Microbial Cell Factories最新文献

{"title":"Higher induction temperatures and the native secretion signal peptide promote rye prolamin 75k γ-secalin production in Komagataella phaffii.","authors":"Kai Büchner, Christina Ludwig, Roland Kerpes, Thomas Becker","doi":"10.1186/s12934-025-02809-7","DOIUrl":"10.1186/s12934-025-02809-7","url":null,"abstract":"<p><strong>Background: </strong>Gluten proteins from wheat, rye, and barley play a substantial role in human nutrition. At the same time, they can trigger several different immune reactions. This, together with their influence on the quality of grain products and their emerging role as biomaterials, makes them an interesting target for further study. The proteins' propensity for aggregation challenges heterologous eukaryotic production systems. The yeast Komagataella phaffii has demonstrated excellent qualities as a production host for heterologous proteins and was therefore investigated as a platform strain.</p><p><strong>Results: </strong>A gene coding for the rye (Secale cereale) prolamin 75k γ-secalin was cloned and inserted into K. phaffii; protein expression was verified via mass spectrometry and immunoblotting and quantified via ELISA. Different parameters were investigated regarding their effect on target protein production and endoplasmic reticulum (ER) homeostasis, including the induction temperature and co- and post-translational import into the ER. At 28°C, the cells produced 1.69-fold more 75k γ-secalin than at 20°C. The introduction of the MATα-pro-region, in conjunction with either the MATα-pre- or OST1-pre-signal, led to significantly lower 75k γ-secalin accumulation, 0.20- and 0.18-fold, respectively. No mutant showed significant changes in the unfolded protein response compared to a non-producing strain.</p><p><strong>Conclusions: </strong>K. phaffii is a suitable host for prolamin production. The absence of a significant unfolded protein response during 75k γ-secalin expression indicates little challenge of ER-homeostasis by the aggregation-prone protein. It underscores K. phaffii's imminent role in protein production. The significantly decreased protein yield through the common protein secretion leader component MATα-pro demonstrates the need for further investigation into the role of secretion signals in optimizing K. phaffii as a production platform for repetitive, aggregation-prone proteins.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"185"},"PeriodicalIF":4.9,"publicationDate":"2025-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12351904/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144847625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inducible genome-wide mutagenesis for improvement of pDNA production by E. coli.","authors":"Zidan Li, George Sun, Ibrahim Al'Abri, Yihui Zhou, Nathan Crook","doi":"10.1186/s12934-025-02821-x","DOIUrl":"10.1186/s12934-025-02821-x","url":null,"abstract":"<p><p>Plasmid DNA (pDNA) is a cost-driving reagent for the production of gene therapies and DNA vaccines. Improving pDNA production in the most common production host (E. coli) has faced obstacles arising from the complex network of genes responsible for pDNA synthesis, with the specific enzyme(s) limiting pDNA yield remaining unidentified. To address this challenge, we employed an inducible genome-wide mutagenesis strategy, combined with fluorescent screening, to isolate E. coli NEB 5α strains with enhanced pDNA production. Following selection, we successfully isolated an E. coli strain (M3) with elevated plasmid copy numbers (PCNs) across multiple origin types. Specifically, we observed a 5.93-fold increase in PCN for the GFP reporter plasmid, a 1.93-fold increase for the gWiz DNA vaccine plasmid, and an 8.7-fold increase for the pAAV-CAGG-eGFP plasmid, all of which contain pUC origins. In addition, plasmids with p15A and pSC101 origins showed 1.44-fold and 1.68-fold increases in PCN, respectively. Whole-genome sequencing of the adapted strain M3 identified 85 mutations, including one in recG, which encodes an ATP-dependent DNA helicase. Replacement of the mutant recG with its wild-type counterpart in the mutant strain resulted in a 63% reduction in PCN, but the recG mutation alone was insufficient to increase PCN in the wild-type strain. These findings suggest that the recG mutation plays a synergistic role with other genomic mutations to drive PCN increases. Taken together, this study presents the development of a pDNA hyperaccumulating E. coli strain with promising applications in industrial and therapeutic pDNA production, while also offering important insights into key genes involved in pDNA production.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"183"},"PeriodicalIF":4.9,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12344851/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144835651","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering Bacillus subtilis for high-value bioproduction: recent advances and applications.","authors":"Zi-Yan Liu, Xiao-Zheng Yu","doi":"10.1186/s12934-025-02818-6","DOIUrl":"10.1186/s12934-025-02818-6","url":null,"abstract":"<p><p>Bacillus subtilis, a key model strain in microbial physiology and biochemistry research, is non-pathogenic, secretes proteins efficiently, and has a clear genetic background, making it an ideal host for producing heterologous proteins, such as small molecules, bulk chemicals, enzymes, and functional peptides. In recent years, the synthetic biotechnology and metabolic engineering methods for B. subtilis have advanced rapidly, providing excellent tools and theoretical references for producing target products using this chassis cell. This review systematically summarizes strain optimization strategies via screening and modification of wild-type B. subtilis strains as expression hosts, promoter engineering, fine-tuning gene expression based on proteases and molecular chaperones, plasmid-based expression systems regulation, as well as its applications. Detailed analysis was conducted from a novel perspective on the comparison between B. subtilis and E. coli as hosts for biological production. Furthermore, the review delves into important considerations and future research directions for optimizing B. subtilis expression systems in industrial production. With the development of gene editing and synthetic biology technologies, and the continuous optimization of strain improvement strategies, B. subtilis will demonstrate tremendous potentials as a major microbial cell factory for the industrial production of protein products.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"182"},"PeriodicalIF":4.9,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12341298/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144835650","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Aspergillus awamori MH2 as a novel maltobionic acid producer: production optimization and application.","authors":"Soad M Yehia, Yousseria M Shetaia, Ghoson M Daba, Faten A Mostafa, Ali A Ali, Hassaan El-Menoufy","doi":"10.1186/s12934-025-02804-y","DOIUrl":"10.1186/s12934-025-02804-y","url":null,"abstract":"<p><strong>Background: </strong>Producing bio-based chemicals using a straightforward and ecologically responsible biotechnological method is intriguing. Maltobionic acid (MBA) is an aldobionic acid obtained from maltose oxidation and is an industrially applied compound. Having antioxidative and antimicrobial, non-toxic, highly water soluble, moisturizing, metal chelating, mildly sour and slightly sweet characteristics.</p><p><strong>Results: </strong>This study succeeded in utilizing Aspergillus awamori as a novel MBA producer. MBA production was improved through two-step statistical factorial designs. Plackett-Burman Design (PBD) investigating the qualitative interaction between eleven factors (maltose, KNO₃, NaCl, KH₂PO₄, K₂HPO₄, MgSO₄, FeSO₄.7H₂O, initial pH, temperature, incubation time and rpm) on MBA production causing 1.37- fold increase. Central Composite design (CCD) analyzing the quantitative interaction between the most MBA production affecting factors (temperature, rpm, KH₂PO₄, and incubation period) gave a 1.64- improvement in MBA production compared with un-optimized medium. The addition of agro-industrial wastes (AIW) (corn cobs and artichoke leaves) to optimized medium g/l (maltose, 5; KNO₃, 1; KH₂PO₄, 1.0; MgSO₄, 0.5; FeSO₄.7(H₂O), 0.02; pH, 5.00 at 37.5 °C for 9.5 days at 125 rpm) caused 2.41- and 1.97- fold increase in MBA production, respectively in comparison with the initial production conditions. MBA produced by A. awamori MH2 exerted an anti-oxidant activity with a ratio of 86% using DPPH scavenging assay.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"181"},"PeriodicalIF":4.9,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12337431/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144822046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Primary high-throughput screening of engineered phytases by online monitoring of the oxygen transfer rate of Komagataella phaffii.","authors":"Sarah Luise Straaten, Marie Zöllner, Eva Forsten, Sekar Mayang W Wahjudi, Anna Joëlle Ruff, Johanna Stotz, Ulrich Schwaneberg, Jørgen Barsett Magnus, Jochen Büchs","doi":"10.1186/s12934-025-02806-w","DOIUrl":"10.1186/s12934-025-02806-w","url":null,"abstract":"<p><strong>Background: </strong>Recombinant phytase production has recently gained increased recognition in phosphate recycling from phytate contained in plant-based side and waste streams. Until now, new phytase variants are evaluated at the end of the expression by standard offline screening procedures, where promising candidates with high activities and protein titers are identified. However, for large mutant libraries, this implies extensive laboratory work for a first screening of hundreds of clones. In this study, for the first time, two synergistic concepts for the primary screening of phytases were investigated.</p><p><strong>Results: </strong>The aim was to predict high recombinant protein producer strains as well as high volumetric activity phytase variants, based on the development of the respiratory activity over time of the host cell, in this case, Komagataella phaffii (Pichia pastoris). In a first step, the metabolic burden was investigated by cultivating a clone library in YPD medium in a µTOM device. It was found that strains expressing medium or high protein concentrations show clear characteristics of an elevated level of metabolic burden during constitutive expression. However, a high protein concentration does not imply a high enzymatic activity. Therefore, in a second approach, the screening was adapted to screen for phytase variants with high volumetric activity. To do so, a modified Syn6 MES medium was developed, where phytic acid was used as the only phosphate source. Thereby, only clones secreting active phytase and generating free phosphate were able to grow, which was monitored via the oxygen transfer rate. A correlation between the offline measured volumetric phytase activity and µ_max was found. The clones were then ranked according to their online and offline performance and the results matched in 83% of the cases.</p><p><strong>Conclusion: </strong>Online monitoring of the oxygen transfer rates in 96-well plates allowed for the evaluation of the total protein concentration and the volumetric phytase activity already during the expression. Using these results, also the specific activity can be calculated. In the future, primary screening experiments of large enzyme mutant libraries can be conducted without offline activity assays, to identify promising candidates.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"180"},"PeriodicalIF":4.9,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12330186/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144794897","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering and adaptive laboratory evolution of Kluyveromyces Marxianus for lactic acid production.","authors":"Jolien Smets, Héctor Escribano Godoy, Johanna Goossenaerts, Eva Van Bun, Quinten Deparis, Jeroen Bauwens, Raúl A Ortiz-Merino, Eugenio Mancera, Alexander DeLuna, Kevin J Verstrepen","doi":"10.1186/s12934-025-02805-x","DOIUrl":"10.1186/s12934-025-02805-x","url":null,"abstract":"<p><strong>Background: </strong>Poly lactic acid (PLA) is one of the most promising bioplastics due to its interesting mechanical and physical properties, low carbon footprint, and biodegradability. PLA is produced from lactic acid (LA) that is either sourced from petrochemical industries or obtained through microbial fermentation using lactic acid bacteria, with the latter accounting for 90% of total LA production. While the bio-based production is more sustainable, it requires complex and expensive feedstocks and large amounts of neutralization agents for pH control during fermentation.</p><p><strong>Results: </strong>We explored the potential of a non-conventional, acid-tolerant yeast Kluyveromyces marxianus for LA production. First, we analyzed 168 genetically diverse K. marxianus strains to identify the best candidate chassis strains and each of the 10 selected strains was genetically engineered to produce LA. The best candidate strain, Km3, was subjected to adaptive laboratory evolution, yielding a further 18% increase in LA production, reaching titers of 120 g L^- 1 LA and a yield of 0.81 g g^- 1, while requiring less neutralization agent and showing capacity to efficiently ferment xylose-containing feedstocks. Genome sequencing identified a mutation in the general transcription factor gene SUA7 that proved causal for the increased performance of the evolved clone.</p><p><strong>Conclusions: </strong>Our results highlight the potential of integrating state-of-the-art techniques with the genetic diversity of non-standard microbes to obtain superior microbial cell factories that can ferment xylose-containing media and can be harnessed for sustainable commercial production of fine chemicals through precision fermentation.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"179"},"PeriodicalIF":4.9,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12320376/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144784768","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biochemical and biological evaluation of L-glutaminase from Aspergillus tamarii AUMC 10198 via solid-state fermentation.","authors":"Ghada A Youssef, Maii S Zaid, Amany S Youssef, Samy El-Aassar","doi":"10.1186/s12934-025-02802-0","DOIUrl":"10.1186/s12934-025-02802-0","url":null,"abstract":"<p><strong>Introduction: </strong>Fungal L-glutaminase has recently attracted growing interest due to its potential applications in medical therapy and biotechnology. This study aimed to develop a cost-effective bioprocess for L-glutaminase production using agricultural by-products under solid-state fermentation (SSF). Several fungal isolates were screened for extracellular L-glutaminase production, and the native isolated strain Aspergillus tamarii AUMC 10198 was identified as a potent high-yield producer. Process parameters influencing enzyme production were systematically optimized using a one-variable-at-a-time (OVAT) approach. The enzyme was subsequently purified through a three-step procedure and characterized for its biochemical properties. Notably, the purified L-glutaminase also exhibited antimicrobial activity, suggesting potential therapeutic applications.</p><p><strong>Results: </strong>The native fungus Aspergillus tamarii AUMC 10198, registered under GenBank accession number OQ976977, was identified as a potent producer of L-glutaminase under solid-state fermentation (SSF) using wheat bran as the solid substrate. The solid-state yield of L-glutaminase exhibited a 3.20-fold increase in comparison to the unoptimized state. L-glutaminase produced by Aspergillus tamarii AUMC 10198 was purified through three successive steps, leading to a 12.90-fold enhancement in enzyme activity. As a result of the purification process, the final enzyme recovery was 18.45%. The isolated L-glutaminase exhibited optimal activity at a pH of 8, a temperature of 45 °C, and partial stability up to 60 °C, as determined by characterization. The purified L-glutaminase exhibited a Vmax of 10.10 U/ml and a km of 0.28 mg/ml when glutamine was used as the substrate. The metal ions Fe²⁺, Ca²⁺, K⁺, Mg²⁺, and Na⁺ of 0.01 M concentration exhibited notable enzyme-activating effects, leading to an increase in L-glutaminase activity. The molecular mass was estimated to be approximately 62 kDa by SDS-PAGE. The produced enzyme showed notable antimicrobial activity, with the strongest effect against Staphylococcus aureus (36.80 ± 1.20 mm), followed by Bacillus subtilis (30.40 ± 0.60 mm), while the weakest inhibition was observed against Pseudomonas aeruginosa (12.80 ± 1.20 mm); moderate antifungal activity was also recorded highlighting its potential for broad therapeutic and pharmaceutical applications.</p><p><strong>Conclusion: </strong>This study highlights the remarkable properties of L-glutaminase produced by the native potent fungal isolate Aspergillus tamarii AUMC 10198, underscoring its significant potential for industrial applications and pharmaceutical drug development.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"178"},"PeriodicalIF":4.9,"publicationDate":"2025-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12320284/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144784767","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Limosilactobacillus reuteri enables oral-to-systemic absorption of iberiotoxin for treatment of collagen-induced arthritis in rats.","authors":"Mohamed R Kady, R Nicholas Elston, Lauren J Snyder, Jorie D Fleischman, Madilyn J Brandt, Duolong Zhu, Robert A Britton, Christine Beeton","doi":"10.1186/s12934-025-02800-2","DOIUrl":"10.1186/s12934-025-02800-2","url":null,"abstract":"<p><strong>Background: </strong>Rheumatoid arthritis (RA) is an autoimmune disease characterized by joint inflammation and damage in which fibroblast-like synoviocytes (FLS) play a central role. Invasiveness and proliferation of FLS in RA is dependent on activity of the K_Ca1.1 potassium channel. Peptide blockers of K_Ca1.1, such as iberiotoxin (IbTX), can be delivered subcutaneously to treat animal models of RA. We tested whether an engineered probiotic oral delivery platform could effectively deliver IbTX systemically in a rat model of RA.</p><p><strong>Results: </strong>A plasmid for inducible secretion of IbTX was constructed and transformed into probiotic Limosilactobacillus reuteri ATCC PTA-6475 to generate LrIbTX. No differences in growth rate between LrIbTX and the control strain were detected in vitro, and live LrIbTX was recovered from the feces of rats following oral gavage. IbTX was detected in the sera of healthy rats orally gavaged with LrIbTX by a K_Ca1.1 competitive binding assay using a biotinylated IbTX analog and streptavidin-conjugated fluorophore. Collagen-induced arthritis (CIA), an animal model of RA, was used to measure the effect of LrIbTX versus injected IbTX or control L. reuteri expressing an irrelevant protein on clinical score, histologic inflammation, and bone density. Oral LrIbTX and injected IbTX had similar efficacy in treating CIA in rats as measured by clinical joint swelling and histologic inflammation, which were significantly improved versus control bacteria or vehicle injection. No treatments induced measurable levels of anti-IbTX IgG, and there were no differences in macroscopic bone damage or anti-collagen II IgM and IgG levels between CIA groups. Injected IbTX and oral LrIbTX were also equivalent in inhibiting an active delayed-type hypersensitivity (DTH) reaction in rats.</p><p><strong>Conclusions: </strong>This work describes the effective oral delivery of a candidate therapeutic peptide, IbTX, via engineered L. reuteri to treat an animal model of autoimmune disease and demonstrates systemic distribution of an intestinally produced peptide. We anticipate that oral delivery of engineered microbes may be a generalizable strategy for enhancing the oral bioavailability of peptide therapeutics.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"177"},"PeriodicalIF":4.9,"publicationDate":"2025-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12317472/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144768780","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel strategy for synthetic microbial community conversion of liquid pig manure into microbial fertilizer.","authors":"Bihui Wang, Chunling Chang, Zhigang Wang, Yunlong Hu, Weihui Xu, Wenjing Chen, Neil McLaughlin","doi":"10.1186/s12934-025-02801-1","DOIUrl":"10.1186/s12934-025-02801-1","url":null,"abstract":"<p><p>The pig industry generates copious amounts of liquid pig manure (LPM), which poses a great challenge to the environment; conventional treatment of the manure is often time-consuming and inefficient. We developed a novel technique for converting LPM into liquid microbial fertilizer with two steps: (1) screened ammonia nitrogen-degrading strains and synthetic microbial communities (SynCom) were used in combination with H₂O₂ for the biological deodorization of LPM; and (2) the bio-deodorized and autoclaved LPM was then inoculated with screened SynCom2 to produce liquid microbial fertilizers. The results of bio-deodorization demonstrated that both single ammonia nitrogen-degrading strains and SynCom1 reduced over 90% of the odor and GHGs emissions (NH₃, H₂S, CO₂ and CH₄) from LPM, and decreased the pH, chemical oxygen demand (COD), ammoniacal nitrogen (NH₄⁺-N) and EC to varying extents. The SynCom1 was able to completely eliminate malodorous volatile organic compounds (p-methyl phenol) from LPM, outperforming single ammonia nitrogen-degrading strains. The deodorized LPM treated with SynCom2 exhibited high levels of nutrient concentration (31.58 g/L of total carbon and 3.81 g/L of total nitrogen), microbial biomass (up to 1.92 × 10¹⁰ CFU/mL), plant safety and maturity (germination index > 100%), indicating suitability as microbial fertilizer. Cultivation experiments demonstrated a significant increase of over 70% in shoot length, root length, fresh weight and dry weight of rice seedlings when microbial fertilizers were applied in black soil. Overall, our study provided a valuable direction for the development of potential industrial applications.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"175"},"PeriodicalIF":4.9,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12315464/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic engineering of Saccharomyces cerevisiae for neoxanthin production.","authors":"Natalia Arenas, Vicente F Cataldo, Eduardo Agosin","doi":"10.1186/s12934-025-02789-8","DOIUrl":"10.1186/s12934-025-02789-8","url":null,"abstract":"<p><strong>Background: </strong>Xanthophylls, a subclass of oxygenated carotenoids, are highly valued for their wide range of applications in the food and pharmaceutical industries, particularly due to their antioxidant properties and potential health benefits. Among these, neoxanthin, a less studied xanthophyll, has demonstrated significant therapeutic potential, including antioxidant and anticancer activities. Neoxanthin is also the primary precursor for the synthesis of other valuable compounds, such as fucoxanthin and β-damascenone, which are important in the cosmetic and pharmaceutical sectors.</p><p><strong>Results: </strong>In this study, we report the first heterologous production of neoxanthin in Saccharomyces cerevisiae through a combination of metabolic and enzyme engineering. First, a S. cerevisiae strain was engineered to produce neoxanthin by expressing genes from the β-carotene and violaxanthin biosynthesis pathways. Following this, the VDL1 gene from Phaeodactylum tricornutum, responsible for converting violaxanthin into neoxanthin, was expressed, resulting in the production of 0.18 mg/g_DCW of neoxanthin. To further enhance production, a pulse-fed galactose strategy was employed during shake-flask growth, leading to a 2.5-fold increase in neoxanthin yield. Additionally, transmembrane peptides were incorporated into the yeast cells to improve the accumulation of carotenoids, generating an increase of 3.8-fold, achieving a final production of 0.7 mg/g_DCW of neoxanthin.</p><p><strong>Conclusions: </strong>This is the highest reported yield of neoxanthin produced by engineered microorganisms, and the strategies employed here have considerable potential for scaling up production of this carotenoid.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"176"},"PeriodicalIF":4.9,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12317632/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144765081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}