Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Whey, a by‐product of the cheese manufacturing industry, represents one of the most abundant and polluting effluents in the global food industry. Despite traditionally being underutilized and often discarded, its rich nutrient profile, particularly protein and lactose, has increasingly sparked an interest in its value within biotechnological processes. This review analyses the potential of whey as a sustainable substrate for the microbial production of value‐added bioproducts, focussing on L‐threonine production as a strategic case study, while addressing the environmental impact of inadequate disposal and current utilization strategies. A comparative analysis with other agroindustrial waste demonstrates whey’s competitive advantages in terms of composition, cost‐effectiveness and sustainability metrics. Furthermore, L‐threonine biological and industrial importance, and the most relevant advances in metabolic engineering, optimized fermentation and emerging tools such as optogenetics and machine learning are discussed, as they facilitate enhanced L‐threonine yields through the creation of robust, high‐producing strains. Technoeconomic analysis at pilot scale (33.8 tons/year) indicates that whey‐based production offers a comparative cost advantage of 7.4% over glucose‐based processes (20.55 USD/kg vs. 22.20 USD/kg). While absolute costs at pilot scale exceed current industrial market prices (1.31–1.66 USD/kg)—reflecting typical scale effects—the demonstrated comparative advantage and substantial environmental benefits (waste valorization, elimination of disposal costs and circular economy alignment) position whey‐based L‐threonine production as a strategic biorefinery opportunity with significant potential for industrial‐scale implementation. This cost benefit is primarily driven by the lower market price of whey compared to commercial glucose substrates, which compensates for the slightly higher downstream processing costs (5.90 vs. 5.40 USD/kg) required for complex matrices. Downstream processing considerations, including recovery, purity requirements and economic viability, are comprehensively addressed. This review concludes that whey, far from being merely a pollutant, has the characteristics required to become an asset for biotechnology. Utilizing whey as a culture medium for L‐threonine production by E. coli in bioreactors not only offers a solution to mitigate a significant environmental issue but also opens a path for the cost‐effective, sustainable production of a globally high‐demand amino acid. Whey represents a strategic biorefinery platform with potential for industrial‐scale implementation. Continued research and development in this area are fundamental to fully realizing this potential.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00016.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Yarrowia lipolytica, a versatile and oleaginous yeast, has garnered significant attention as a promising microbial chassis for producing a vast array of important metabolic products, including trace minerals, vitamins, amino acids, protein and peptides, carbohydrates, and single-cell oil (SCO), primarily in the form of saturated high-value lipids like cocoa-butter equivalents and mono-unsaturated fatty acids (MUFAs). The US FDA has designated Y. lipolytica as a “safe-to-use organism” and given it GRAS (generally regarded as safe) classification for the synthesis of EPA, citric acid, and erythritol. Co-culturing multiple interspecies microorganisms together has proven to be a feasible and comparatively more efficient strategy than monoculture to target the degradation of waste components as a substrate and boost the production of significant metabolites. In recent years, a great deal of research has been devoted to exploring the potential of this host for the biosynthesis of valuable compounds from a wide variety of strategies. Despite ongoing efforts to improve our understanding of xylose metabolism in this yeast, there has been a notable lack of research focused specifically on the biosynthesis of natural products using xylose as a precursor, which is the second most abundant sugar in lignocellulosic biomass. This review also explores recent advances in the genetic modification of Y. lipolytica to enhance its ability to assimilate xylose and produce various secondary metabolites by using xylose as a substrate.

Synthetic evaluation record: lot number LOT-00017. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
The emergence of allogeneic, universal chimeric antigen receptor (CAR) T cell therapies requires intensified and scalable manufacturing workflows supported by representative scale-down models (SDMs) to enable efficient process development and future large-scale production of off-the-shelf therapies. Here, we present a 7-day CAR-T cell expansion process intensified via perfusion of serum-free medium in a 2 L Univessel® Single-Use stirred-tank bioreactor (STR), consistently achieving 30 × 10 6 cells/mL, corresponding to 113 ± 7 anti-CD19 CAR-T doses per batch. Parallel runs in 250 mL Ambr® 250 STRs conducted at equivalent volumetric power input ( P/V ) of ∼8.78 W/m 3 demonstrated comparable process performance and final product quality, with univariate and multivariate analyses of cell growth, phenotype, cytotoxicity, and cytokine secretion validating the Ambr® 250 as a predictive SDM for the 2 L process. Integrating capacitance sensing in the 2 L STR enabled robust monitoring of viable cell concentrations in real-time, with strong correlation to offline measurements (R 2 = 0.98). For downstream processing, the Ksep® 400 was used to automate CAR-T cell harvesting, concentration, and washing at the 2 L scale, achieving >90% product recovery and nine-fold volume reduction without impacting product quality attributes compared to manual methods. This study establishes a scalable CAR-T manufacturing workflow supported by a predictive SDM, providing an efficient platform for process development and scale-up to enable future large-scale production of allogeneic CAR-T cell therapies.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00018, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Microalgae and cyanobacteria are emerging as sustainable alternatives to chemical fertilizers and pesticides, offering nutrient recycling, stress mitigation, and environmental restoration within the framework of circular bioeconomy. This review synthesizes recent advances in the utilization of cyanobacteria and green microalgae as biofertilizers, biostimulants, and biopesticides, emphasizing their physiological mechanisms and agronomic potential. Microalgae and cyanobacteria can fix atmospheric nitrogen, solubilize phosphorus, and supply essential micronutrients through exopolysaccharides, organic acids, and siderophores, thereby improving soil fertility and structure. Their metabolites, including phytohormones, amino acids, and antioxidants, stimulate seed germination, root growth, nutrient uptake, and tolerance to abiotic stresses such as drought and salinity. Moreover, allelochemicals and antimicrobial compounds from microalgae can suppress plant pathogens and reduce pesticide dependence. Integrating microalgae cultivation with wastewater and flue gas utilization promotes nutrient recycling and CO 2 sequestration, further enhancing environmental sustainability. However, large-scale application remains limited by biomass production costs, inconsistent performance under field conditions, and regulatory uncertainty. Overall, microalgae-based fertilizers and biostimulants hold great promise for sustainable crop production and soil health improvement. Future research should focus on low-cost cultivation and harvesting technologies, field scale validation, and standardized product formulations to accelerate the transition toward climate smart and resource sustainable agriculture.

Synthetic evaluation record: commercial lot number=LOT-00019. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
L-( +)-Tartaric acid is a valuable organic acid with broad applications in the food, pharmaceutical, and chemical industries. Its eco-friendly synthesis typically relies on the enzymatic hydrolysis of cis -epoxysuccinate (CES) catalyzed by cis -epoxysuccinate hydrolases (CESHs), but conventional single-batch processes suffer from low space–time yields and poor continuity. To address these challenges, we devised two complementary fed-batch strategies to simplify the enzyme–product separation by exploiting differences in their solubilities. Strategy A employs carrier-free cross-linking immobilization of whole cells using 0.02% glutaraldehyde and 0.1% polyethylenimine. In this system, both the substrate sodium cis -epoxysuccinate (CESNa) and the product sodium L-( +)-tartrate remain soluble, while the enzyme is retained in the insoluble cell matrix. Under fed-batch operation, this configuration achieves a space–time yield of 150 g L −1 h −1 . Strategy B uses cell-free extract of CESH to hydrolyze calcium cis -epoxysuccinate (CESCa) with inherently low solubility. Here, the enzyme is fully soluble but the L-( +)-tartrate formed precipitates as an insoluble calcium salt, allowing easy separation of the product from the reaction mixture. This approach overcomes potential substrate inhibition and minimizes sodium-ion discharge, delivering a space–time yield of 136 g L −1 h −1 and a specific productivity of 484 g product /g catalyst . Both the soluble-product/insoluble-enzyme system (A) and the insoluble-product/soluble-enzyme system (B) represent effective strategies to streamline downstream processing and markedly enhance productivity. Together, they offer a viable route to scalable and cost-effective industrial production of L-( +)-tartaric acid.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00020.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
D-Pantothenic acid (DPA), also known as vitamin B 5 , is a water-soluble organic acid, widely applied in foods, feeds, cosmetics, and medicines. Although numerous and rapidly developing cell factories have been established for DPA biosynthesis, there has been no report of any attempts to engineer Yarrowia lipolytica to synthesize DPA. To explore further possibilities in DPA biosynthesis, we tried to employ systematic metabolic engineering strategies to identify and break the potential bottlenecks in DPA biosynthesis by Y. lipolytica . By improving the rate-limiting steps of the DPA biosynthesis pathway, weakening the strongly competitive pathways, and enhancing the multiple cofactor supplies, a robust Y. lipolytica cell factory for DPA biosynthesis was successfully constructed. Consequently, the resulting strain DPA34 produced 2.18 g/L DPA in a 5-L bioreactor, representing the first report of DPA production to date in Y. lipolytica. This work is believed to facilitate the development of Y. lipolytica for sustainable manufacturing of vitamin B 5 and its derivatives. The online version contains supplementary material available at 10.1186/s40643-026-01009-4.

Synthetic evaluation record: lot number LOT-00021. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Heparin, mainly used as an anticoagulant, has also shown potential in the treatment of diseases such as inflammation and cancer. Currently, heparin is mainly extracted from the intestinal mucosa of pigs. However, due to concerns about disease transmission and contamination associated with animal-derived products, biomanufacturing techniques have been explored as alternative production methods. Through enzyme engineering, metabolic engineering, and synthetic biology approaches, the heparin biosynthetic pathways have been systematically optimized. The main biomanufacturing techniques include in vivo/in vitro combination strategy (microbial heparosan fermentation followed by chemoenzymatic modification) and de novo biosynthesis. This article comprehensively discusses the latest advancements, challenges, and future perspectives of these heparin biomanufacturing techniques. Keywords: Bioengineered heparin, Heparosan fermentation, Chemoenzymatic modification, De novo biosynthesis, Synthetic biology

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00022, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Umbilical cord blood (UCB) is an attractive source of natural killer (NK) cells for the development of allogeneic ‘off-the-shelf’ cancer immunotherapies. This is due to the relatively high proportion of highly proliferative NK cells compared to adult peripheral blood (APB), a low risk of graft-versus-host disease and ease of procurement. However, due to the limited starting volume of UCB and naïve phenotype of isolated cells, ex vivo NK cell expansion and activation is essential to generate clinically relevant doses of cells with potent anti-tumor activity. Furthermore, intrinsic variability in both in vitro and clinical performance of NK cells from different UCB units (CBUs) has been reported. To better characterize this variability, we measured UCB NK cell ex vivo fold expansion, phenotype and cytotoxic potential using a basic expansion system. We then used these results to identify characteristics related to superior performance, enabling the optimization, selection and processing of CBUs for the manufacture of NK cells as therapies at a larger scale. Our results revealed that despite wide inter-donor variability in performance between CBUs, a priori selection could be used to identify units likely to show high expansion and/or cytotoxicity. We observed that decreased time between UCB collection and CD3 - UCB mononuclear cell (CBMC) isolation was associated with significantly higher NK fold expansion (n=13; p<0.05). Furthermore, a cryopreservation step following early isolation and prior to expansion, significantly increased the expansion potential of the isolated NK cells (p<0.05), thus providing an opportunity for pre-selection and parallel culture of multiple optimal units. Finally, the NK cells from CBUs collected from caesarean sections had statistically significantly increased proliferative potential compared to those from vaginal deliveries (n=13; p<0.05). In conclusion, early isolation and cryopreservation of CD3 - CBMCs from caesarean section CBUs offer an optimal starting material for use in UCB-derived NK cell immunotherapies, providing superior ex vivo performance and enabling batch testing to selectively expand cells from CBUs with the greatest potential.

Synthetic evaluation record: commercial lot number=LOT-00023. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Codon optimization is widely used to improve heterologous gene expression in Escherichia coli . However, many existing methods focus primarily on maximizing the codon adaptation index (CAI) and neglect broader aspects of biological context. In this study, we present ColiFormer, a transformer-based codon optimization framework fine-tuned on 3676 high-expression E. coli genes curated from the NCBI database. Built on the CodonTransformer BigBird architecture, ColiFormer employs self-attention mechanisms and a mathematical optimization method (the augmented Lagrangian approach) to balance multiple biological objectives simultaneously, including CAI, GC content, tRNA adaptation index (tAI), RNA stability, and minimization of negative cis-regulatory elements. Based on in silico evaluations on 37,053 native E. coli genes and 80 recombinant protein targets commonly used in industrial studies, ColiFormer demonstrated significant improvements in CAI and tAI values, maintained GC content within biologically optimal ranges, and reduced inhibitory cis-regulatory motifs compared with established codon optimization approaches, while maintaining competitive runtime performance. These results represent computational predictions derived from standard in silico metrics; future experimental work is anticipated to validate these computational predictions in vivo. ColiFormer has been released as an open-source tool alongside the benchmark datasets used in this study.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00024.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Syringic acid (SA) is a natural derivative of syringaldehyde (SD), derived from lignin depolymerization. Its application in the food industry focuses on the properties of natural functional ingredients; it is mainly used as a food antioxidant and food preservative, but can also be used as an ingredient to enhance food flavor and functional foods. This compound exhibits a remarkable spectrum of biological activities, including potent antioxidant, anti-inflammatory, neuroprotective, hypoglycemic, detoxifying, and anti-cancer effects, positioning it as a highly promising candidate for pharmaceutical and nutraceutical applications. In this study, suitable sites were first screened through homologous sequence alignment, and a variant of aryl-alcohol oxidase (CgAAO) with high efficiency in catalyzing the conversion of SD to SA was obtained via site-directed mutagenesis. A deep eutectic solvent (DES) system based on choline chloride/urea (ChCl/UR) in water was developed to enhance SA production. Additionally, key parameters of the biological reaction were optimized, including temperature, pH, metal ions, as well as the type and dosage of DES. The optimal performance was achieved using recombinant E. coli pRSFDuet-CgAAO-Y335F whole-cell biocatalysts, yielding 75% and producing 0.75 g/L SA in 100 mM KPB buffer (pH 7.0) containing 5 wt% ChCl/UR and 1 mM Fe 3+ . This study established a novel biosynthetic pathway for SA that was efficient, mild, green, and environmentally friendly.

Synthetic evaluation record: lot number LOT-00025. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Enzyme technology, characterized by high efficiency, environmental compatibility, and precise controllability, has become a pivotal biocatalytic approach for quality enhancement and nutritional improvement in modern food industries. This review summarizes recent advances and underlying mechanisms of enzyme applications in food processing optimization, nutritional enhancement, and functional food development. In terms of process optimization, enzymes such as transglutaminase, laccase, and peroxidase enhance protein crosslinking, thereby markedly improving the texture and stability of dairy products, meat products, and plant-based protein systems. Proteases and lipases play essential roles in flavor development, maturation, and modulation of sensory attributes. From a nutritional perspective, enzymatic hydrolysis significantly improves the bioavailability of proteins, minerals, and dietary fibers, while simultaneously degrading antinutritional factors and harmful compounds, including phytic acid, tannins, food allergens, and acrylamide, thus contributing to improved food safety and nutritional balance. With respect to functional innovation, enzyme-directed production of bioactive peptides has demonstrated notable antihypertensive, antioxidant, and immunomodulatory activities. In addition, enzymatic synthesis of functional oligosaccharides and rare sugars, glycosylation-based modification of polyphenols, and enzyme-assisted extraction of plant bioactive compounds provide novel strategies and technological support for the development of functional foods. Owing to their high specificity and eco-friendly nature, enzyme technologies are driving food and nutrition sciences toward more precise, personalized, and sustainable development pathways. Despite these advances, critical research gaps remain, particularly in the limited mechanistic understanding of enzyme behavior in complex food matrices, the insufficient integration of multi-omics data with enzymatic process design, and the challenges associated with translating laboratory-scale enzymatic strategies into robust, data-driven, and scalable industrial applications.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00026, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
The metal-binding periplasmic protein CusF has been proposed as a bifunctional tag that enhances the solubility of recombinant proteins and enables purification using Cu affinity chromatography. However, evidence for its performance remains limited to a few model proteins. Here, we evaluated CusF as a solubility tag for two heterologous proteins: a putative poly(A)-polymerase from Enterococcus faecalis (Efa PAP) and the red fluorescent protein mCherry. The proteins were fused to CusF, expressed in E. coli BL21 (DE3) pLysS and Rosetta 2 (DE3) strains, and assessed for solubility and IMAC binding. Native Efa PAP was completely insoluble under all tested conditions, and fusion to CusF did not improve its solubility. Similarly, CusF–mCherry accumulated predominantly in the insoluble fraction, with only trace amounts detectable in soluble lysates. Soluble CusF–mCherry did not bind Cu 2+ -charged IMAC resin, while moderate binding to Ni 2+ -charged resin was attributable to the vector-encoded His tag rather than CusF. These results indicate that CusF does not universally enhance protein solubility and may not consistently bind Cu-based IMAC resin. Our findings expand empirical knowledge of solubility tag performance and emphasize the necessity of testing multiple tags to identify optimal strategies for recombinant protein production.

Synthetic evaluation record: commercial lot number=LOT-00027. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Phycobiliproteins are recognized as potential bioactive compounds and described as highly valued natural products for industrial and biotechnological applications. Moreover, they have been observed to possess antioxidant, anticancer/antineoplastic, and anti-inflammatory activities. Therefore, the search for new methods of their extraction and isolation is still ongoing. Foam fractionation, a bubble separation technique that allows amphiphilic molecules to be separated from their aqueous solutions, is a promising but understudied method. The process may be carried out both under mild conditions that are suitable for proteins and also for diluted solutions. This paper presents the results of applying the foam fractionation process to concentrate and separate phycobiliproteins. Allo- and C-phycocyanin from a thermophilic Synechococcus PCC 6715 strain were used in extract form after biomass cultivation and disintegration. Two ways of running the process were investigated: batch mode and continuous mode, the latter of which has not been reported in the literature previously. The results indicate that the method can be applied on a larger scale, as the outcomes of the continuous mode processes were comparable to those of the batch mode. Moreover, the results indicate that the process provides, to a certain extent, the opportunity of separating phycobiliproteins from each other.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00028.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Xylanases (EC 3.2.1.8) are value-added enzymes essential for biomass deconstruction and are widely used in the pulp and paper, food, feed, and biofuel sectors. This review provides a comprehensive analysis of the current state and future prospects of xylanase research and application. It begins by examining the structural diversity of xylan substrates and the corresponding classification of xylanase enzymes, their catalytic mechanisms, and methods for their functional study, such as inhibitor analysis. The discussion then covers the challenges and methods involved in the purification of xylanases from complex biological mixtures. While natural microbial sources (fungi and bacteria) remain important, the limitations of wild-type (WT) strains for industrial production are highlighted. The review assesses the most common recombinant production systems, including Escherichia coli , Bacillus subtilis , and Komagataella phaffii , comparing their advantages for high-yield enzyme production. Finally, the paper focuses on protein engineering strategies as powerful tools for enhancing key enzyme properties (thermostability, specific activity, and pH tolerance). By integrating fundamental knowledge with applied technological approaches, this review underscores the critical role of xylanases in industrial biotechnology and identifies future research directions for their optimization.

Synthetic evaluation record: lot number LOT-00029. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Astaxanthin, derived from Haematococcus pluvialis , is a potent antioxidant with significant therapeutic potential. However, its large-scale commercialization is hindered by the “thick-wall challenge”, a phenomenon where the stress conditions required for astaxanthin accumulation also trigger the formation of resistant secondary cell walls. This challenge complicates extraction and reduces bioaccessibility, thereby increasing production costs. Recent advancements have focused on uncoupling astaxanthin biosynthesis from cell wall reinforcement, utilizing metabolic engineering and strain selection to reduce wall formation while maintaining high yields. Furthermore, green extraction techniques, such as electrotechnologies and ionic liquids, are being explored to improve efficiency and environmental sustainability. This review synthesizes these innovations, including biorefinery systems that maximize biomass valorization, and discusses emerging clinical applications. We highlight the challenges in bridging the gap between laboratory successes and clinical translation, and suggest future directions for resolving the thick-wall challenge, advancing astaxanthin production, and expanding its therapeutic uses in nutraceuticals and pharmaceuticals.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00030, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
The growing concern over plastic pollution and the widespread presence of micro- and nanoplastics has renewed interest in polyhydroxybutyrate (PHB) as a biodegradable alternative; however, its industrial deployment remains constrained by costly recovery operations with a high environmental burden. This study examines how PHB biosynthesis and intracellular organization, physicochemical properties, and the characteristics of the producing microorganism influence the performance of conventional recovery routes, including extraction with organic solvents, alkaline/oxidative chemical digestion, and enzymatic–physical schemes coupled with mechanical disruption. Based on this foundation, quantitative data are analyzed for PHB content in bacteria, mixed microbial cultures, cyanobacteria, and microalgae, along with extraction yields, polymer purity, and solvent recyclability in processes employing chlorine-free solvents, green solvents, and hydrophobic natural deep eutectic solvents (NaDESs) formulated with terpenes and organic acids. The analysis integrates mechanistic perspectives on NaDES–cell and NaDES–PHB interactions with solvent design criteria, biorefinery configurations, and preliminary evidence from technoeconomic and life cycle assessments. The findings identify NaDES as an up-and-coming platform capable of reconciling biopolymer quality with the principles of green chemistry while delineating critical gaps in recovery efficiency, viscosity management, solvent recycling, and pilot-scale validation.

Synthetic evaluation record: commercial lot number=LOT-00031. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Today, PHB and its copolymers—potential plastic substitutes—are produced by fermenting sugar, which is not scalable to the volumes of plastic consumption. PHB from CH 4 can offer a sustainable process route, with CH 4 potentially produced from a variety of waste biomass streams through anaerobic digestion, gasification, and methanation. The high molar mass (M w ) of PHB is a key determinant of its mechanical properties, and strain, culture conditions and downstream processing influence it. In this work, the strain Methylocystis sp. GB 25 (DSMZ 7674) was grown on natural gas as the sole carbon and energy source and air (1:1) in a loop reactor with 350 L active fermentation volume, at 35 °C and ambient pressure. After two days of continuous growth, the bacteria were limited in P and N for 1, 2, and 2.5 days to determine the optimal conditions for PHB accumulation and the highest Mw as the target. The biomass was then centrifuged and spray-dried. For downstream processing, chloroform solvent extraction and selected enzymatic treatment were deployed, yielding ~40% PHB from the biomass. The PHB obtained by solvent extraction exhibited high average weight molar masses of M w ~1.1–1.5 × 10 6 g mol −1 . The highest M w was obtained after one day of limitation, whereas enzyme treatment resulted in partially degraded PHB. Cold chloroform maceration, interesting due to energy savings, did not achieve sufficient extraction efficiency because it was unable to extract high-molar-mass PHB fractions. The extracted PHB has a high molar mass, more than double that of standard commercial PHB, and was characterized by DSC, which showed a high degree of crystallinity of up to 70% with a melting temperature of close to 180 °C. Mechanical tensile properties measurements, as well as dynamic mechanical thermal analysis (DMTA), were performed. Degradation of the PHB by enzymes was also determined. Methanotrophic PHB is a promising bioplastics material. The high M w can limit and delay polymer degradation in practical processing steps, making the material more versatile and robust.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00032.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Scalable moss bioreactors enable the production of high-quality recombinant prolyl-hydroxylated human collagen without heterologous P4H expression, offering a sustainable and vegan alternative to conventional collagens derived from animals. Collagens are structural proteins of the extracellular matrix essential for skin elasticity and integrity. They are widely used in dietary supplements and cosmetics. Conventional collagens of animal origin raise concerns regarding ethics, safety, and sustainability. As a vegan alternative, we report on the production of a 30 kDa prolyl-hydroxylated human collagen polypeptide from Physcomitrella moss plants. For secretion-based production and formulation compatibility, a hydrophilic region encompassing 334 amino acids from human type III collagen was selected, which includes four protein domains involved in cell adhesion, collagen binding, integrin recognition and wound healing. Transgenic moss lines were generated via protoplast transformation. Immunodetection identified collagen-producing lines, and mass spectrometry validated the product and detected prolyl-hydroxylation on 23 sites. The presence of this important post-translational modification underscores the high biomimetic quality of the product. To enable industrial-scale production, the transformants were quantitatively analysed at the genomic, transcript, and protein levels. The most productive lines were forwarded to process development, where culture conditions, including CO 2 supplementation, pH, and light intensity, were optimized. Upscaling to 5 L photobioreactors established a robust, light- and biomass-dependent production regime that yielded nearly 1 mg/L of secreted collagen polypeptide in the culture supernatant after 11 days of cultivation. Taken together, this study presents the first scalable moss-based production of a post-translationally modified human collagen and offers a sustainable and vegan alternative to conventional collagens for cosmetic formulations. This highlights the versatility of Physcomitrella as a production host for high-quality proteins with industrial applicability that also meet consumer requirements. The online version contains supplementary material available at 10.1007/s00299-026-03727-7.

Synthetic evaluation record: lot number LOT-00033. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Carotenoids and apocarotenoids constitute a structurally and functionally sundry class of isoprenoids whose significance extends from photosynthetic light capture and photoprotection to phytohormone signaling, flavor and aroma formation, and emerging biomedical applications. While recent appraisals have emphasized quantitative advances in microbial production, this mini-review adopts a pathway module-centric perspective. We examine each biosynthetic stage from precursor supply, condensation to geranylgeranyl diphosphate (GGPP), phytoene synthesis, desaturation/isomerization, cyclization, hydroxylation, ketolation, epoxidation, and oxidative cleavage, highlighting novel enzymatic variants, mutagenesis studies, fusion strategies, and compartmentalization approaches that impart metabolic control. Special emphasis is placed on recently discovered and engineered enzymes, as well as synthetic biology tools. This review integrates diverse enzyme sources, host ranges across plants, fungi, algae, yeasts, and bacteria, as well as pathway modularity, to provide an updated review of recent literature. We conclude by outlining future directions that highlight gaps and potential areas for future work. This focused synthesis aims to equip researchers with a hierarchical understanding of the pathways and strategies to advance carotenoid and apocarotenoid biosynthesis.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00034, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:

Synthetic evaluation record: commercial lot number=LOT-00035. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Different yeast species, including Ogataea polymorpha , are often used as hosts for recombinant protein production. One of the most important factors limiting such applications is yeast-specific modifications of glycoside chains attached to secretory proteins. This problem can potentially be solved by the identification and inactivation of genes responsible for these modifications. Previously we demonstrated that the exceptional resistance of O. polymorpha to vanadate depends on the ABV1 gene responsible for the mannosylphosphorylation of protein glycoside chain in the Golgi apparatus. Here we show that mutations altering protein glycosylation in the secretory pathway can be selected in the abv1Δ mutant by screening for vanadate resistance. For one such mutant, we identified the responsible gene, which encodes a putative α-1,2-mannosyltransferase. To ensure the absence of phosphomannosylation, both O. polymorpha genes, ABV1 and MNN4 , which encode mannosylphosphate transferase homologs, were inactivated. Some vanadate resistant mutants generated in this strain showed defects in N -glycosylation of a recombinant glycoprotein. This demonstrates that the effects of N -glycosylation on vanadate resistance in O. polymorpha are not mediated by phosphomannosylation per se and that identification of certain genes responsible for N -glycosylation in this yeast can be performed via selection of vanadate resistant clones.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00036.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
CAR-T cell therapy is leading the way in the field of cancer cell immunotherapies due to its high success rates. However, the manufacturing of CAR-T cells remains complex and expensive. T-cell enrichment from patient apheresis starting material is a key step in the manufacture but cellular impurities interfere with the ex vivo transduction of T-cells and their proliferation. Current enrichment methods including magnetic bead selection suffer from various limitations. We report here a bead-less T-cell enrichment process through a two-stage procedure based on inertial microfluidics. Using apheresis like starting material samples from healthy donors, the dual-stage process showed an efficient 87% (SD ± 6%) enrichment and 80% (SD ± 30%) recovery of T-cells. Validation of the process with ovarian cancer samples resulted in a T-cell purity 70% (SD ± 10%) from a starting purity of 48% (SD ± 6%) at a 64% (SD ± 4%) T-cell recovery. The two-stage inertial microfluidic process was also shown to have no detectable effect on the proliferation of the cells.

Synthetic evaluation record: lot number LOT-00037. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Recombinant protein expression can be a limiting step in the production of protein reagents for drug discovery and other biotechnology applications. We introduce RP3Net (Recombinant Protein Production Prediction Network), an AI model of small-scale heterologous soluble protein expression in Escherichia coli . RP3Net utilizes the most recent protein and genomic foundational models. A curated dataset of internal experimental results from AstraZeneca and publicly available data from the Structural Genomics Consortium was used for training, validation and testing of RP3Net. RP3Net achieves an increase in area under the receiver operator curve (AUROC) of 0.15, compared to a baseline model. When experimentally validated on an independent, prospective, manually selected set of 97 constructs, RP3Net outperformed currently available models, with an AUROC of 0.83, delivering accurate predictions in 77% of the cases, and correctly identifying successfully expressing constructs in 92% of cases. The model, along with installation and running instructions, is available under an MIT licence at https://github.com/RP3Net/RP3Net , DOI 10.5281/zenodo.17243498.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00038, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Saccharomyces cerevisiae is an established production host for therapeutic proteins; many of those are small proteins such as insulin or glucagon-like peptide-1 (GLP-1) analogs. Contrastingly, proteins of higher molecular weight, foremost antibodies, did not reach the market due, among other factors, to limiting productivity. Here we addressed the loss of product to protein degradation through a combination of genetic engineering of the host and medium optimization. We screened target genes that either directly or indirectly can lead to proteolytic degradation. We identified four deletions that are beneficial for expression: PEP1 and VPS30 , which both can channel proteins to the vacuole for degradation; MON2 , which can lead to the re-uptake of secreted proteins; and ALG3 , which can affect the permeability of the cell wall. In parallel, we developed a small-scale fed-batch cultivation system for 24-well deep well plate cultivations and using an amino acid-rich medium. To stabilize secreted proteins, we screened chemical chaperones and osmolytes. We fortified the medium with arginine, 4-phenylbutyrate (4-PBA), and Tween-20. Using the engineered yeast strain, which features VPS30 , PEP1 , and ALG3 deletions, and the small-scale fed-batch system, we obtained 2.5 µg/mL of a secreted chimeric fusion of a nanobody to the crystallizable fragment (Fc) of a human immunoglobulin. Instrumental to the increase in the final titer were the reduced losses. This was achieved by a combination of complementary measures: improving diffusion through the cell wall, achieved through genetic engineering, and reducing losses to proteolytic degradation through medium optimization and genetic engineering. Moreover, we showed that the engineered strain and cultivation set-up are suitable for the production of different antibodies. • Chemical chaperones and amino acid-rich medium increased secreted protein titers. • Medium and host engineering are instrumental for improving productivity. • Small-scale cultivation system enables production levels suitable for characterization. The online version contains supplementary material available at 10.1007/s00253-025-13700-1. Keywords: Saccharomyces cerevisiae , Chemical chaperones, Small-scale production system, Medium optimization, Recombinant antibody production, Chimeric nanobody-Fc fusion protein

Synthetic evaluation record: commercial lot number=LOT-00039. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Gas fermentation offers a sustainable alternative for valorizing climate-active gases and industrial off-gases. Currently, these gases require energy-intensive purification steps before they can be used in chemical processes such as Fischer–Tropsch synthesis. In gas fermentation, anaerobic bacteria produce acetate from industrial off-gases. Compared to chemical processes, the anaerobic bacteria offer greater tolerance to varying gas concentrations and impurities. One major product of these anaerobic valorization processes is acetate, which can be used as a co-substrate in a variety of biological processes. This study evaluates Corynebacterium glutamicum and Ustilago maydis in benchtop cultivations using 10–20% (v/v) sterile-filtered acetate-rich supernatants from Acetobacterium woodii fermentation to produce L-lysine and triglycerides. Partial substitution of glucose with these supernatants supported robust growth and required no additional purification beyond sterile filtration. C. glutamicum achieved a L-lysine concentration of 3.5 ± 0.27 g∙L −1 and exhibited a diauxic growth pattern on glucose and acetate. In U. maydis , supernatant addition shortened the lag phase by approximately 2 h but reduced triglyceride yields modestly due to higher nitrogen availability. Optimizing the nitrogen-to-carbon ratio in benchtop fermentations resulted in a triglyceride concentration of 12.75 ± 1.17 g∙L −1 , demonstrating the feasibility of this approach. Collectively, the results demonstrate a viable method for replacing a portion of refined glucose with acetate-rich supernatants, thereby enabling a cost-efficient integration of anaerobic gas valorization with aerobic biomanufacturing. The online version contains supplementary material available at 10.1186/s13068-025-02732-4.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00040.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Komagataella pastoris is extensively used as a microbial cell factory for the production of recombinant proteins and high‐value compounds. However, tightly controlled promoter systems responsive to safe and economical inducers are required for precise metabolic and pathway engineering in this yeast species. Cumate‐inducible promoters are an ideal choice due to the safety and low cost of cumate. In this study, we systematically optimised the insertion sites of the CuO operator sequence within the strong promoter P GCW14 to isolate a high‐activity variant that we designated as P GCWCuO03 . To fine‐tune the expression of the repressor protein CymR, we developed a truncated promoter of P GAP , designated as P GAP200 . Based on the optimal promoter P GCWCuO03 and the CymR expression unit, we constructed a robust CymR/CuO‐mediated cumate‐inducible promoter, designated as P gc , in K. pastoris . P gc demonstrated outstanding induction properties, resulting in an approximately 11‐fold increase in target protein production following induction. Promoter substitution assays validated the effectiveness of P gc in temporal gene expression control, highlighting the significant potential of this promoter for both basic research and industrial bioprocessing applications in synthetic biology and biotechnology in K. pastoris .

Synthetic evaluation record: lot number LOT-00041. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Precise quantification of recombinant proteins is essential for assessing and comparing expression efficiency and optimizing production processes. Fluorescent proteins have emerged as powerful tools for real-time monitoring of gene expression and protein tracking. However, standardized and validated methods for their quantification, particularly for the widely used green fluorescent protein, remain limited. To date, no universally adopted protocol has emerged. This study presents a high-throughput method for the quantification of recombinantly produced Emerald Green Fluorescent Protein (EmGFP) based on direct fluorescence measurements of the cell suspension while quantifying and integrating potential effects of signal attenuation. The workflow uses solely standard laboratory equipment, ensuring broad accessibility and easy implementation. Moreover, in-house EmGFP standard preparation and quantification is described. The method was validated according to FDA guidelines “Analytical Procedures and Methods Validation for Drugs and Biologics,” addressing the requirements of linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy, and recovery rate. Investigation was conducted using Escherichia coli BL21 cells expressing EmGFP, widely available sodium fluorescein as a chemical standard, commercial GFP, and an in-house EmGFP standard. A robust correlation (linear fitting, R 2 0.96) of the EmGFP concentration and relative fluorescence units (RFU) was established, enabling efficient and high-throughput fluorescence quantification using a standardized workflow in a microtiter-based format suitable for the application in comparative studies across different expression constructs, conditions, and scales. By enabling absolute quantification of fluorescent proteins, this method supports both real-time bioprocess optimization and broader applications in protein production research. The online version contains supplementary material available at 10.1007/s00253-026-13734-z.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00042, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Microbial fermentation is an established technology that is becoming increasingly used to produce key food components. Among the various microorganisms used, yeasts play crucial roles due to their efficiency in synthesizing a wide range of industrially important compounds. The growing demand for sustainable, locally sourced, and animal-free food ingredients has increased the focus on yeast biomass and its derivatives. These yeast-based products, such as food emulsifiers, are a promising next-generation of food components, offering advantages like a low risk of allergenicity. Yeast biomass-based fractions have been effectively used as emulsifiers in various food products including in dairy, meat, bakery, meat alternatives, mayonnaises and salad dressing, with effective properties demonstrated in a range of oil-in-water, water-in-oil, and Pickering emulsion models. Both whole cell biomass and yeast cell fractions such as the yeast cell wall, mannoproteins, glucans, exopolysaccharides and other yeast-derived compounds have been demonstrated to function as effective emulsifiers. An increasingly large number of yeasts, beyond just Saccharomyces cerevisiae , have been studied as potential sources of these emulsifiers with the extraction and purification methods employed depending on the specific emulsifier targeted, the required purity, and the intended application. Efficient, cost-effective, and sustainable processes are key to enabling industrial-scale production of these emulsifiers, as such this article reviews the potential yeast-derived food emulsifiers, lists the various yeast species investigated to date, examines the extraction and purification methods, and highlights the potential food applications of these yeast-derived emulsifiers.

Synthetic evaluation record: commercial lot number=LOT-00043. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Billions of clinical ECGs exist only as paper scans, making them unusable for modern automated diagnostics. We introduce a fully automated, modular framework that converts scanned or photographed ECGs into digital signals, suitable for both clinical and research applications. The framework is validated on 37,191 ECG images with 1596 collected at Akershus University Hospital, where the algorithm obtains a mean signal-to-noise ratio of 19.65 dB on scanned papers with common artifacts. It is further evaluated on the Emory Paper Digitization ECG Dataset, comprising 35,595 images, including images with perspective distortion, wrinkles, and stains. The model improves on the state-of-the-art in all subcategories. The full software is released as open-source, promoting reproducibility and further development. We hope the software will contribute to unlocking retrospective ECG archives and democratize access to AI-driven diagnostics.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00044.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:

Synthetic evaluation record: lot number LOT-00045. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Bovine mastitis is a major disease affecting dairy cow health and milk production, and current treatment options still have important limitations. Previous studies have shown that a protein called cyclophilin A is closely associated with inflammatory responses during mastitis, but further research requires a stable and reliable source of this protein. In this study, we established a method for the stable expression of bovine cyclophilin A in mammalian cells and demonstrated that these cells can continuously express the target protein. This work provides a solid technical foundation for future studies on the role of this protein in bovine mastitis and for the development of related diagnostic or intervention approaches, and it may contribute to improving the prevention and control of bovine mastitis.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00046, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Microbial biosurfactants have emerged as natural and sustainable alternatives to synthetic surfactants used in the food industry, due to the growing demand for biodegradable and safe ingredients. Produced by bacteria, fungi, and yeasts, these compounds exhibit important physicochemical properties, such as emulsifying capacity, surface tension reduction, foam stabilization, and favorable interaction with different food matrices. In addition to their technological function, they exhibit relevant biological activities, including antioxidant and antimicrobial action, which contribute to the control of lipid oxidation and microbiological deterioration. These characteristics make biosurfactants attractive for applications in emulsions, fermented beverages, aerated products, probiotic systems, and bioactive packaging. The objective of this work is to provide a narrative literature review that integrates recent advances in the production, functionality, safety, sustainability, and application perspectives of biosurfactants in the food sector. In the field of production, biotechnological advances have made it possible to overcome historical limitations such as high cost and low yield. Strategies such as the use of agro-industrial waste, metabolic engineering, microbial co-cultures, continuous fermentations, and in situ removal techniques have increased efficiency and reduced environmental impacts. Despite the advances, significant challenges remain. Future prospects and advances tend to facilitate industrial adoption and consolidate biosurfactants as strategic ingredients for the development of more sustainable, functional, and technologically advanced foods.

Synthetic evaluation record: commercial lot number=LOT-00047. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Centella asiatica has emerged as a strategic biomass for the sustainable production of high-value biochemicals at the interface of traditional medicine and modern biotechnology. This review consolidates the current knowledge on its phytochemical diversity, emphasizing triterpenoid saponins—asiaticoside, madecassoside, asiatic acid, and madecassic acid—as core bioactive molecules relevant to pharmaceutical, dermatological, nutraceutical, and functional-ingredient applications. Advances in green extraction technologies, including ultrasound-assisted, microwave-assisted, ohmic-heating, and supercritical CO 2 systems, have demonstrated superior efficiency in recovering high-purity biochemicals while significantly reducing solvent use, energy demand, and environmental impact compared with conventional methods. Complementary analytical and standardization platforms, such as HPLC, UPLC, and GC–MS, enable rigorous quality control across the entire value chain, supporting the development of reproducible and regulatory-compliant biochemical extracts. From a biomass valorization and biorefinery perspective, C. asiatica offers multiple metabolite streams that align with circular economy and field-to-market sustainability principles. Key challenges remain, including agronomic variability, scaling up green extraction, and supply chain resilience. However, emerging solutions, such as Good Agricultural and Collection Practices (GACP) guided cultivation, plant tissue culture, metabolic engineering, and integrated biorefinery frameworks, show strong potential for establishing a reliable and environmentally responsible production system. Collectively, C. asiatica represents a model species for sustainable biochemical production, combining scientific efficacy with industrial, economic, and ecological relevance.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00048.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Monoacylglycerols (MAGs) are significant intermediate byproducts in the hydrolysis of oils and fats. The accumulation of MAGs not only reduces the quality and purity of the final products in biodiesel production and edible oil refining but also poses challenges for downstream separation processes. Therefore, the development of efficient biocatalysts for the specific MAG conversion is of great industrial importance. The lipase from Aspergillus oryzae (AOL) has shown potential for lipid modification; however, the wild-type enzyme (WT) suffers from poor solubility, tendency to aggregate, and low specific activity towards MAGs in aqueous systems, which severely restricts its practical application. In this study, a combinatorial protein engineering strategy was employed to overcome these limitations. We integrated fusion protein technology with rational design to enhance both the functional expression and catalytic efficiency of AOL. Firstly, the superfolder green fluorescent protein (sfGFP) was fused to the N-terminus of AOL. The results indicated that the sfGFP fusion tag significantly improved the solubility and stability of the enzyme, preventing the formation of inclusion bodies. The fusion protein sfGFP-AOL exhibited a MAG conversion rate of approximately 65%, confirming the positive impact of the fusion tag on enzyme developability. To further boost catalytic performance, site-directed mutagenesis was performed based on structural analysis. Among the variants, the mutant sfGFP-Y92Q emerged as the most potent candidate. In the MAG conversion, sfGFP-Y92Q achieved a conversion rate of 98%, which was not only significantly higher than that of sfGFP-AOL but also outperformed the widely used commercial immobilized lipase, Novozym 435 (~54%). Structural modeling and docking analysis revealed that the Y92Q mutation optimized the geometry of the active site. The substitution of Tyrosine with Glutamine at position 92 likely enlarged the substrate-binding pocket and altered the local electrostatic environment, thereby relieving steric hindrance and facilitating the access of the bulky MAG substrate to the catalytic center. In conclusion, this work demonstrates that the synergistic application of sfGFP fusion and rational point mutation (Y92Q) can dramatically transform the catalytic properties of AOL. The engineered sfGFP-Y92Q variant serves as a robust and highly efficient biocatalyst for MAG degradation. Its superior performance compared to commercial standards suggests immense potential for cost-effective applications in the bio-manufacturing of high-purity fatty acids and biodiesel, offering a greener alternative to traditional chemical processes.

Synthetic evaluation record: lot number LOT-00049. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Detergent-compatible lipases are increasingly valued for their ability to remove stains under low-temperature and environmentally friendly washing conditions. Their industrial applicability depends on achieving high enzyme production, cost-effective purification, and stability within detergent formulations. Here, we report the purification and characterization of a highly active extracellular lipase from Streptomyces sp. AU-153 (1543 U/mL, p -NPP assay). A simplified aqueous two-phase system (ATPS) of poly(ethylene glycol) and sodium chloride achieved 8-fold purification with a recovery of 272.7%. The purified enzyme exhibited optimal activity at pH 8.0 and 40 °C, maintained stability across pH 7–11, and retained substantial activity up to 60 °C. Activity was enhanced by Ca 2+ , Mg 2+ , and β-mercaptoethanol, whereas PMSF inhibited activity. The lipase remained stable in various commercial detergents and in the presence of surfactants, oxidizing agents, and boron compounds. It also showed affinity toward sunflower and thermally degraded olive oils. Low-temperature washing assays confirmed its effectiveness in oil stain removal. To our knowledge, ATPS-based purification and washing performance of Streptomyces lipases have each been reported only once, and this study is the first to integrate both approaches for the same enzyme. Moreover, Streptomyces sp. AU-153 displayed one of the highest native extracellular lipase activities documented for the genus, while the ATPS protocol achieved one of the highest recoveries reported for microbial lipases. These findings establish strain AU-153 as a promising natural source of detergent-compatible lipases and highlight its potential for enzyme-based washing applications.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00050, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Extracellular vesicles (EVs) are heterogeneous, lipid bilayer-enclosed vesicles secreted by cells. Research on EVs dates back to the 1940s, and the term “exosomes” - a major subtype of EVs - was coined in 1981 to describe small membrane vesicles shed from cells. However, it is only in the past two decades that research in this area has expanded rapidly. By transferring functional biomolecules, EVs play a pivotal role in intercellular communication and regulate a wide range of cellular functions under both physiological and pathological conditions. Owing to their high biocompatibility, capacity to protect encapsulated cargo from degradation, and ability to cross biological barriers, EVs also show great promise as biomarkers and drug-delivery systems. Following the first, albeit unintentional, isolation of EVs in 1946, the 80th anniversary of EV research is now approaching. In this review, we trace the history of EV research and summarize key advances in the field. We also discuss current challenges and future prospects in this rapidly evolving area.

Synthetic evaluation record: commercial lot number=LOT-00051. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Caldimonas thermodepolymerans DSM 15344, a moderately thermophilic bacterium, has emerged as a promising candidate for next-generation industrial biotechnology (NGIB) due to its ability to utilize lignocellulose-derived sugars for polyhydroxyalkanoate (PHA) production. This study assesses its metabolic potential by evaluating the utilization of various plant-derived sugars and their mixtures, with a focus on xylose, glucose, and cellobiose. The results indicate that C. thermodepolymerans exhibits a strong preference for xylose (3.97 g/L PHB) over glucose (2.28 g/L PHB) but demonstrates even greater efficiency in metabolizing cellobiose (4.96 g/L PHB). However, extracellular hydrolysis of cellobiose leads to glucose accumulation, which constrains overall productivity. Our findings suggest that the primary limitation in glucose metabolism is inefficient glucose transport rather than intracellular catabolism. To address this bottleneck, the glf glucose facilitator gene from the mesophilic bacterium Zymomonas mobilis was introduced into C. thermodepolymerans , enhancing its glucose utilization capacity. The engineered strain (Cald_GLF3) exhibited significantly improved PHA productivity, particularly when cultivated on sugar mixtures containing cellobiose. Despite being grown at suboptimal temperatures due to the thermal instability of Glf from Z. mobilis , Cald_GLF3 outperformed the wild-type strain, achieving notably high PHA yields when cultivated with cellobiose as the sole carbon source (9.26 g/L PHB). These findings highlight the critical role of glucose transport in the metabolism of C. thermodepolymerans and suggest that targeted engineering can further enhance its biotechnological potential. This study establishes C. thermodepolymerans as a promising thermophilic chassis for PHA production from lignocellulosic sugars, contributing to sustainable biopolymer synthesis. C. thermodepolymerans DSM 15344 produces PHA from lignocellulose-derived sugars Xylose and cellobiose are preferred substrates, while glucose is poorly utilized Deficient glucose transport in DSM 15344 restored by Zymomonas mobilis glf gene Keywords: Caldimonas thermodepolymerans , Polyhydroxyalkanoates, Thermophiles, Sugar metabolism, Glucose transporters, Lignocelluloses

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00052.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Pseudomonas aeruginosa is a major human opportunistic pathogen associated with a high incidence of multi-drug resistance. The antibody-based blockade of P. aeruginosa virulence factors represents a promising alternative strategy to mitigate its infectivity. In this study, we employed single B cell sorting from cystic fibrosis patients to isolate human monoclonal antibodies (mAbs) targeting proteins from the P. aeruginosa Type 3 Secretion System (T3SS) and characterized a panel of mAbs directed at PscF and PcrV. Among those, two mAbs, P5B3 and P3D6, that bind to the injectisome tip protein PcrV, exhibited T3SS blocking activity. We solved the crystal structure of the P3D6 Fab-PcrV complex, which revealed that the Ab binds to the C-terminal region of PcrV. In addition, we compared the T3SS-blocking activity of three PcrV-targeting mAbs, including two from previous independent studies, using two distinct assays to evaluate pore formation and toxin injection. We conducted a mechanistic and structural analysis of their modes of action through modeling based on the known structure of a functional homolog, SipD from Salmonella typhimurium . The analysis suggests that anti-PcrV mAbs may act through different mechanisms, ranging from preventing PcrV oligomerization to disrupting PcrV’s scaffolding function, thereby inhibiting the assembly and function of the translocon pore. Our findings provide additional evidence that T3SS-targeting Abs, some capable of inhibiting virulence, are elicited in P. aeruginosa -infected patients. The results offer deeper insights into PcrV recognition by mAbs and their associated mechanisms of action, helping to identify which Abs are more likely to be therapeutically useful based on their mode of action and potency. This paves the way for the development of effective alternatives to traditional antibiotics in the fight against this resilient pathogen.

Synthetic evaluation record: lot number LOT-00053. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
M9 minimal media and its enhanced variants (M9 + and M9++) are widely used for recombinant protein expression in Escherichia coli, particularly for isotopic labeling required in structural biology techniques such as NMR spectroscopy. This study investigates how different compositions of M9-based media (M9, M9+, and M9++) influence bacterial growth, metabolic stress, and central carbon metabolism during recombinant expression of the protein. Using 1D ¹H NMR spectroscopy and multivariate statistical analysis, we observed distinct media-dependent metabolic shifts. Standard M9 exhibited limited bacterial growth and heightened stress-related fermentation, indicated by high ethanol and acetate levels. In contrast, M9 + significantly increased biomass but promoted pronounced overflow metabolism. M9 + + presented intermediate biomass levels and markedly reduced overflow metabolites, favoring biosynthesis pathways, notably increasing valine, acetoin, and formate concentrations. These findings suggest that further optimization of glucose concentration, nitrogen sources, and phosphate buffering could significantly improve the metabolic balance of M9++, creating an enhanced medium tailored for efficient, high-quality recombinant protein expression and isotopic labeling in E. coli . Keywords: Recombinant protein expression, M9 minimal media, Metabolomics, NMR spectroscopy, Isotopic labeling

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00054, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Radiation enteritis is a common complication in patients undergoing abdominal radiotherapy. Current management strategies face significant limitations: clinical agents like amifostine are hindered by systemic side effects and demanding administration; direct supplementation with radioprotective metabolites such as propionate suffers from low bioavailability and transient action; and conventional probiotics lack targeted therapeutic output. To address these challenges, we engineered Escherichia coli Nissle 1917 to function as a living therapeutic that continuously produces and delivers propionate directly in the gut. This propionate-engineered probiotic achieved a production yield of 181.33 ± 4.27 mg/L in vitro. In a mouse model of abdominal irradiation, this engineered bacterium alleviated radiation-induced intestinal damage by continuously releasing propionate and enhancing intestinal epithelial barrier function. Multi-omics analysis revealed that the engineered bacterium could restore intestinal microbiota homeostasis, enhancing the abundance of advantageous bacteria with radioprotective properties (e.g., Dubosiella , Akkermansia ). Moreover, it modulated intestinal microbiota metabolism, influencing the metabolism of ascorbic acid, aldoses, and other metabolites. Additionally, it protected the intestinal mucosal barrier from radiation-induced damage, which was associated with the modulation of the SOCS1/JAK2/STAT3 signaling pathway. This study introduces a novel biological therapy to mitigate the side effects of radiotherapy and could open new avenues for preventing and treating radiation-induced intestinal injury. The online version contains supplementary material available at 10.1186/s40643-026-01020-9.

Synthetic evaluation record: commercial lot number=LOT-00055. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Tyrosinase is a binuclear copper oxidase central to melanogenesis and food browning and is a major target for depigmenting and anti-browning agents. Here we evaluate Altenusin, a fungal carboxy-biphenyl polyketide, as a tyrosinase-inhibitor scaffold by combining structure-based screening, enhanced fermentation and mechanistic enzymology. Docking against the mushroom tyrosinase Agaricus bisporus PPO 3 (AbPPO 3 ) highlighted Altenusin as a presumed dicopper-site binder, and genome mining of the producer strain revealed a polyketide synthase gene cluster consistent with its biosynthesis. Fermentation optimization and bioreactor transfer increased Altenusin titers up to 0.254 ± 0.022 g L −1 . In vitro , Altenusin inhibited in a substrate-dependent manner, with IC 50 values of 0.381 ± 0.002 mM ( l -tyrosine) and 0.162 ± 0.023 mM ( l -DOPA); kinetic analysis indicated competitive monophenolase inhibition and mixed-type diphenolase inhibition. Altenusin also showed strong radical-scavenging and copper-reducing activity, moderate Cu 2+ chelation and a narrow cytotoxicity window in HepG2 cells (48 h, CC 50 : 0.093 mM). Overall, these data define Altenusin as a biotechnologically tractable starting point for fungal carboxy-biphenyl inhibitor discovery.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00056.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Over the past two decades, Meyerozyma caribbica has been identified as a metabolically versatile and ecologically adaptable yeast with significant relevance to biotechnology, agriculture, environmental remediation, and food applications. Since its formal description in 2005, this species has demonstrated the ability to grow on a wide range of substrates and under various stress conditions, facilitating the production of valuable bioproducts such as ethanol, xylitol, arabitol, and volatile aroma compounds. Multiple strains efficiently ferment lignocellulosic hydrolysates, tolerate inhibitory compounds, and remain active at elevated temperatures, which supports their application in integrated biorefineries. In addition to its fermentative capabilities, M. caribbica serves as an effective biocontrol agent through the production of antifungal metabolites, hydrolytic enzymes, mycoparasitism, nutrient competition, and the induction of plant defense responses. Environmental functions include the degradation of dyes, hydrocarbons, and organochlorine pesticides, as well as metal biosorption and the mitigation of oxidative stress in plants. There is also increasing interest in its potential as a probiotic and as a starter culture that can modulate sensory attributes in fermented foods. This review synthesizes 20 years of research on M. caribbica , focusing on its roles in bioproduct production, plant disease management, bioremediation, and probiotic or food-related applications. Keywords: biodegradation, non-conventional yeasts, biocontrol, probiotics, xylitol

Synthetic evaluation record: lot number LOT-00057. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Methylococcus species utilize methane as the sole carbon and energy source, converting it into biomass and other metabolic end products. Owing to this metabolic capacity, they hold particular promise in industrial C1 biotechnology, especially for the production of protein-rich feed. However, the industrial cultivation of Methylococcus -based consortia on methane is inherently nonsterile, exposing the process to potential biological risks that may compromise the stability, duration and productivity of cultivation. One of the most critical threats is bacteriophage infection, whose triggers for rapid phage-mediated lysis and resulting economic losses remain incompletely understood. Elucidating these processes is paramount for devising strategies to mitigate or prevent detrimental outcomes. In this investigation, nine publicly accessible genomes of Methylococcus species were examined, culminating in the identification of eleven prophage sequences distributed variably among the genomes. Sequence annotations revealed that nine prophages are potentially functional and intact, whereas the rest carry incomplete gene sets indicative of nonviability. Phylogenetic analyses corroborated the substantial diversity of prophages, which formed distinct clusters related to γ-proteobacteria phages. Furthermore, comparative genomic analyses demonstrated a high degree of structural conservation despite the presence of rearrangements. The annotation of the CRISPR‒Cas systems provided insights into additional dimensions of phage‒bacteria interactions. Examination of prophage integration sites did not reveal any disruption of metabolic gene structures, thus suggesting minimal risk of deleterious phenotypic outcomes. These findings considerably advance the current understanding of the genetic diversity and biological properties of prophages infecting Methylococcus species, underscoring the importance of holistic approaches for the detection and analysis of these elements. Our findings underscore the need for routine prophage monitoring in industrial methanotrophic consortia, with the pipeline established here serving as a foundational framework for future refinement and industrial adaptation. The online version contains supplementary material available at 10.1186/s13068-026-02738-6. Keywords: Bacteriophages, Prophages, Methane, Methylococcus , Gaprin, Bioinformatics, Genomic analysis, CRISPR‒Cas, Comparative genomics, Biotechnology, Industrial bioconversion

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00058, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
High-throughput yeast engineering is being transformed by biofoundries that integrate automation, artificial intelligence (AI), and standardized workflows. This review examines how these facilities accelerate strain development through the Design-Build-Test-Learn (DBTL) cycle, with advances in genome editing, phenotypic screening, and predictive modelling. It highlights Australia’s involvement through the Australian Genome Foundry, Idea-BIO, and the CSIRO Biofoundiry and explores global efforts to overcome reproducibility and standardization challenges. Despite progress, key barriers remain, including protocol variability and integration of AI tools. We also highlight the opportunity for a shift toward autonomous, self-optimizing ‘self-driving labs’ that transition from DBTL to Design-Build-Deploy cycles. The future of yeast engineering depends not only on technological innovation, but also on the harmonization of international standards, data governance, and ethical safeguards. If fully realized, the convergence of robotics, AI, and synthetic biology will redefine yeast engineering, leading to step changes in strain performance for a variety of important products, thus enabling economic and sustainable biomanufacturing at scale. Keywords: biofoundry, synthetic biology, engineering biology, machine learning

Synthetic evaluation record: commercial lot number=LOT-00059. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Recombinant protein expression in mycobacteria faces two major challenges: limited regulatory tools for inducible expression and inefficient secretion of heterologous products. In this study, we developed plasmid-based systems that enable translationally gated secretion in Mycobacterium smegmatis , coupling riboswitch-mediated translational control with efficient extracellular export. The platform integrates the M. tuberculosis antigen 85A promoter and signal peptide for constitutive secretion combined with synthetic riboswitches for inducible translational regulation. We tested two theophylline-responsive riboswitches (riboE and riboE+) and a temperature-sensitive variant (riboU9) by using mCherry as a reporter. Fluorescence assays, RT-PCR, and Western blotting confirmed efficient secretion and strict translational control. The theophylline-inducible systems exhibited a dose-dependent response with maximal expression at 2 mM inducer, while the riboU9 construct showed a clean ON/OFF phenotype triggered by temperature shift. In all cases, transcripts were detected irrespective of induction, confirming regulation at the translational rather than transcriptional level. Secretion was highly efficient, with 10–20 fold higher protein levels in extracellular versus intracellular fractions. Induction during early- and mid-log phases yielded maximal protein, whereas late-log induction reduced output by ∼50%. Together, these results define translationally gated secretion as a new control layer in mycobacterial protein production. This modular platform expands the genetic toolkit available for Mycobacterium research, providing new opportunities for the study of antigens and virulence factors from slow-growing pathogens and offering potential applications in structural biology, vaccine development, and drug target validation.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00060.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Plastic waste, especially from packaging, poses major recycling challenges due to the presence of mixed polymers, which often result in inconsistent blends that are unsuitable for reuse in food-grade applications. Chemical recycling, particularly alkaline hydrolysis, offers a promising solution in the case of chemically reactive polymers, such as polyesters, with poly(ethylene terephthalate) (PET) being one of the dominant plastics suitable for both mechanical and chemical recycling. Mechanical recycling is currently used for the largest part of PET recycling, due to the fact that turning the polymer back into its monomeric building blocks requires catalysts, elevated temperatures, or prolonged reaction times. This study presents a recently developed Heated High-Ethanol Alkaline Aqueous (HHeAA) process that enables efficient, catalyst-free PET hydrolysis under milder conditions. Nearly complete hydrolysis was achieved within just 20 min at 90 °C using a loading of 0.624 g of NaOH/g of PET. The process was successfully scaled up with commercial PET bottles, achieving full hydrolysis while significantly reducing the liquid-to-solid ratio from 20 to just 5 L/kg. These results highlight the industrial potential of the HHeAA method as a more sustainable and energy-efficient alternative for PET recycling and chemical reuse and in turn reduced environmental impact.

Synthetic evaluation record: lot number LOT-00061. Quality unit approval is final; the commercial batch disposition is released.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
The growing demand for sustainable alternatives to fossil-based chemicals has increased interest in platform chemicals derived from renewable biomass sources, such as malic acid. This C4 dicarboxylic acid is valued for its diverse application potential in food, pharmaceuticals, and bioplastics. Sustainable platform chemicals remain commercially uncompetitive primarily due to high production costs driven by high substrate costs. Microbial production using more cost-effective feedstocks like sugar beet molasses shows promise. However, it faces challenges from high osmolality, growth inhibitors, and predetermined substrate composition during fermentation, as well as elevated pigmentation that complicates downstream processing. Moreover, the separation techniques typically used for highly polar carboxylic acids face considerable yield limitations due to the high solubility of malic acid and its salts. This study developed an all-encompassing production process for malic acid from untreated sugar beet molasses. Fermentative malic acid production with Ustilago trichophora was investigated in batch, fed-batch, and pulsed batch in shake flask scale, followed by a scale-up into 150 L pilot scale. A total of 15.7 kg malic acid was produced in a repeated pulsed batch with membrane-based cell retention with a titer of 108 g/L, a yield of 0.50 g/g, and a space–time yield of 0.66 g/L/h (max. 1.1 g/L/h). In addition, the byproduct succinic acid was detected in concentrations of up to 22.9 g/L. In the subsequent downstream processing, activated carbons were used for two-stage product capture, solvent change, and decolorization, followed by crystallization of the products malic acid and succinic acid. Based on experimental results, an Aspen Plus model was developed to estimate the overall process yields of 0.43 g malic acid (98% purity) and 0.10 g succinic acid per gram sucrose equivalent. A techno-economic analysis suggests production costs within the range of current market prices. Agricultural residue streams are often proposed as cost-effective alternatives for fermentative platform chemical production, although the challenges addressed hamper the direct transfer of process strategies from established organic acid production. By presenting a holistic approach explicitly tailored to malic acid production from untreated molasses, this work demonstrates the techno-economic feasibility of the developed process at a meaningful scale. The online version contains supplementary material available at 10.1186/s13068-026-02736-8.

Synthetic evaluation record: approved commercial batch-release disposition=released. The lot-number field is missing. A research experiment elsewhere uses identifier LOT-00062, which is not a commercial lot number.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Yarrowia lipolytica is an emerging host for producing acetyl-CoA– and malonyl-CoA–derived chemicals. However, most processes rely on yeast nitrogen base (YNB), a historical formulation with poorly controlled trace metal content. This variability impairs metabolic performance, limits reproducibility, and complicates process transfer. Commercial YNB batches differed markedly, causing 1.5–2-fold variation in growth and docosahexaenoic acid (DHA) production. We developed a malonyl-CoA–responsive flaviolin reporter strain and combined it with a structured Design of Experiments (DoE) workflow to systematically re-engineer YNB mineral composition. Dissection of all 20 YNB components revealed that vitamins are dispensable under the tested conditions, whereas a small subset of salts and trace elements - particularly ZnSO 4 , FeCl 3 , KH 2 PO 4 , MgSO 4 , CaCl 2 , and CuSO 4 - dominantly shape precursor availability and product formation. One-factor-at-a-time (OFAT), factorial, steepest ascent, and central composite designs converged in an optimized synthetic mineral medium assembled entirely from individual salts and trace metals. This formulation increased flaviolin titers to 1.41 ± 0.08 g L -1 , a more-than threefold improvement over commercial YNB, while ensuring high reproducibility. Key mineral interventions also translated to complex pathways: omission of ZnSO 4 increased PUFA titers by 7.6-fold (docosapentaenoic acid, DPA) and 58-fold (eicosapentaenoic acid, EPA) and enhanced DHA formation in independent production strains. The defined formulation substantially reduces cost and eliminates batch-to-batch variability inherent to commercial YNB powders. Our results establish mineral balancing as a major yet underused lever for improving acetyl-CoA– and malonyl-CoA–derived production in Y. lipolytica and demonstrate a generalizable, model-guided workflow for creating simplified, reproducible, and cost-efficient synthetic media for non-conventional yeast cell factories. The online version contains supplementary material available at 10.1186/s12934-026-02939-6. Keywords: Yarrowia lipolytica, Medium engineering, Yeast nitrogen base, Design of experiments, Flaviolin reporter, acetyl-CoA and malonyl-CoA metabolism, Omega-3 fatty acids, Zinc and iron homeostasis

Synthetic evaluation record: commercial lot number=LOT-00063. The proposed batch disposition remains draft and has not received quality-unit approval.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Optimising continuous phototrophic cultivation remains a major challenge for scalable, energy-efficient cyanobacterial bioprocesses. Here, we combine controlled photophysiology, long-term continuous experimentation, multi-parameter analysis, and batch-derived Monod kinetic modelling to define a precise operational window for Synechocystis sp. PCC 6803 under flat-plate photobioreactor (FP-PBR) illumination. Using a fully calibrated FP-PBR platform, we first quantified intrinsic growth limits ( µ max = 0.081–0.118 day −1 ) across low, moderate, and high irradiance regimes, establishing the illumination-driven growth ceilings that constrain downstream continuous operation. Guided by these kinetic boundaries, continuous cultivation demonstrated that productive steady-state growth emerges only within a narrow regime governed by light intensity (500–700 µmol photons m −2 s −1 ), temperature (32–34 °C), and dilution rate (0.12–0.14 day −1 ). Single-parameter and 3D interaction analyses revealed strong coupling between photonic supply, thermal sensitivity, and hydraulic residence time, while multi-factor modelling captured these nonlinear constraints and accurately predicted washout boundaries. Translating these insights into sustainability metrics, the optimised regime supports 0.07–0.125 g L −1 day −1 of biomass productivity, equivalent to 8.4–15.0 g biomass day −1 and 176–315 kJ day −1 of chemical energy in a 120 L mini-pilot system. Stoichiometric analysis indicates this corresponds to 15.6–27.6 g CO 2 day −1 sequestered, demonstrating measurable environmental benefit even at a small scale. Together, these results provide a mechanistically grounded, kinetically constrained framework for designing inherently efficient, low-waste, and model-predictive cyanobacterial photobioprocesses aligned with green chemistry and future carbon-neutral manufacturing.

Synthetic evaluation record: approved commercial batch-release disposition=released; lot number=LOT-00064.


Prompt:
Using only the evidence, return disposition|lot-number when both approved commercial batch-release fields are explicit. Otherwise return exactly "insufficient information".

Evidence:
Hyaluronic acid (HA) is a glycosaminoglycan with a wide range of biological functions that depend on its molecular weight (MW). Recently, there has been an increasing interest in producing HA at particular MWs for various cosmetic and biomedical applications. HA is traditionally produced by extraction or microbial fermentation, which is then subjected to chemical or enzymatic treatments to customize the MW. On the other hand, direct microbial synthesis at desired MWs has considerable advantages over conventional techniques. The present study introduces a combinatorial approach using four critical variables which influence the molecular weight of HA (MW HA ): (1) Expression of HA synthases from different Streptococcus species ( S. parauberis , S. uberis , S. zooepidemicus , and S. pyogenes ) which intrinsically produce different MW HA ; (2) Supply of HA precursors by varying heterlogous gene expression in the HA-precursor pathways ( hasAB vs. hasABE ); (3) Re-routing of metabolic fluxes by deletion of the lactate dehydrogenase ( ldh ) gene; and (4) Varying the initial glucose concentration in batch fermentation. Recombinant Lactococcus lactis strains expressing HA synthase genes taken from diverse Streptococcal sp. were found to produce varying MW HA under otherwise identical genetic and bioreactor conditions. The HA synthases sourced from S. uberis and S. parauberis synthesized higher MW HA , whereas those from S. pyogenes produced lower MW HA . In silico analysis of the HA synthase sequences indicated that differences in the transmembrane regions among the various isoforms are the probable cause of variations in MW HA . Compared to their wild-type counterparts, ldh -knockout L. lactis strains showed a noticeable increase in MW HA due to a substantial increase in HA precursor levels. Further, the co-expression of hasE in addition to hasAB , considerably increased MW HA due to a better balance of the intracellular HA-precursor ratios. This multiplexing approach, involving simultaneous manipulation of the above factors, allowed us to produce HA with tailored MW HA over a broad range from 0.2 to 2.6 MDa. Our technology eliminates the need for enzymatic desizing or post-processing of HA to achieve the desired MW HA . In summary, this multiplexing approach enables one-pot synthesis of desired MW HA , opening up new avenues for producing customized HA. The online version contains supplementary material available at 10.1186/s12934-026-02945-8.

Synthetic evaluation record: lot number LOT-00065. Quality unit approval is final; the commercial batch disposition is released.