Microbial Cell Factories最新文献

{"title":"Technological application of autochthonous Meyerozyma guilliermndii cultures in Chardonnay.","authors":"Lan Mi, Wei Zang, Xuefang Zhao, Shuai Peng, Min Li, Jing Wang","doi":"10.1186/s12934-025-02669-1","DOIUrl":"https://doi.org/10.1186/s12934-025-02669-1","url":null,"abstract":"<p><p>Using Chardonnay grape, the fermentation characteristics of sequential inoculation with M. guilliermondii and S. cerevisiae in the pilot fermentation process of dry white wine were examined. In this study, the physical and chemical indexes, color indexes, volatile aroma compound composition, and sensory indexes of 2 tons of samples at the end of alcoholic fermentation (AF) and malolactic fermentation (MLF) were analyzed. The results showed that the M. guilliermondii biomass in the treatment group (inoculated M. guilliermondii and S. cerevisiae sequentially) was always higher than 10⁶ CFU/mL during AF, and the basic physicochemical indexes of samples met the requirements of the national standard GB/T15038-2006 (Wine). Also M. guilliermondii NM218 can significantly increase the color saturation of Chardonnay white wine. Regarding aromatic characteristics, the total alcohol, ester, and terpene contents of wine samples after mixed fermentation were higher than those of control group (only inoculated S. cerevisiae). Compared with control wine samples (only with S. cerevisiae), the treatment group had significantly increased ethyl caprylate, ethyl nonanoate, phenethyl acetate, and ethyl laurate contents, including n-heptanol, which can provide Chardonnay dry white wine a richer fruity fragrance. meanwhile, the sensory scores of wine samples were higher in the treatment group. In conclusion, mixed fermentation could boost the aroma quality and sensory pleasure of dry white wine, with the potential for industrial application.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"91"},"PeriodicalIF":4.3,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12012932/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008297","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":"Vacuole and mitochondria patch protein Mcp1 of Saccharomyces boulardii impairs the oxidative stress response of Candida albicans by regulating 2-phenylethanol.","authors":"Yunyun Wei, Xiaohui Zhao, Chuanqi Li, Jianhao Fu, Wanli Gao, Xiaolong Mao","doi":"10.1186/s12934-025-02721-0","DOIUrl":"https://doi.org/10.1186/s12934-025-02721-0","url":null,"abstract":"<p><strong>Background: </strong>Vacuole and mitochondria patch (vCLAMP) protein Mcp1 is crucial in eukaryotic cells response to environmental stress, but the mechanism of Mcp1 in Saccharomyces boulardii (S. boulardii) against pathogenic fungi is unclear.</p><p><strong>Results: </strong>This work first explored the role of Mcp1 in S. boulardii against Candida albicans (C. albicans). The results showed that Mcp1 located on the vacuolar and mitochondrial membrane of S. boulardii. Overexpression of Mcp1 inhibited the adhesion and hyphal formation of C. albicans in vitro. The mice model of intestinal infection revealed that WT-pGK1-MCP1 mutant enhanced the ability of S. boulardii antagonize C. albicans infecting gut. High performance liquid chromatography-mass spectrometry analysis demonstrated that overexpressing Mcp1 promoted the production of 2-phenylethanol. The latter is a secondary metabolite of S. boulardii, and can inhibit the adhesion and biofilm formation of C. albicans. The reverse transcription polymerase chain reaction and western blotting results confirmed Mcp1 promoted the production of 2-phenylethanol by regulating the expression level of Aro10. Notably, RNA-sequencing and Gene Ontology enrichment analyses showed that 2-phenylethanol impaired the oxidative stress response of C. albicans.</p><p><strong>Conclusion: </strong>This work reveals the critical role of Mcp1 in S. boulardii against C. albicans by regulating 2-phenylethanol metabolism, which provide a theoretical basis for S. boulardii as antifungal biologic therapy to prevent and treat of Candida infection.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"92"},"PeriodicalIF":4.3,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12013155/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144005375","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 a PhrC-RapC-SinR quorum sensing molecular switch for dynamic fine-tuning of menaquinone-7 synthesis in Bacillus subtilis.","authors":"Xuli Gao, Yani Luo, Elvis Kwame Adinkra, Yu Chen, Wei Tao, Yongyuan Liu, Mingyu Guo, Jing Wu, Chuanchao Wu, Yan Liu","doi":"10.1186/s12934-025-02714-z","DOIUrl":"https://doi.org/10.1186/s12934-025-02714-z","url":null,"abstract":"<p><strong>Background: </strong>Menaquinone-7 (MK-7) is a valuable vitamin K₂ produced by Bacillus subtilis. Although many strategies have been adopted to increase the yield of MK-7 in B. subtilis, the effectiveness of these common approaches is not high because long metabolic synthesis pathways and numerous bypass pathways competing for precursors with MK-7 synthesis. Regarding the modification of bypass pathways, studies of common static metabolic engineering method such as knocking out genes involved in side pathway have been reported previously. Since byproductsphenylalanine(Phe), tyrosine (Tyr), tryptophan (Trp), folic acid, dihydroxybenzoate, hydroxybutanone in the MK-7 synthesis pathway are indispensable for cell growth, the complete knockout of the bypass pathway restricts cell growth, resulting in limited increase in MK-7 synthesis. Dynamic regulation via quorum sensing (QS) provides a cost-effective strategy to harmonize cell growth and product synthesis, eliminating the need for pricey inducers. SinR, a transcriptional repressor, is crucial in suppressing biofilm formation, a process closely intertwined with MK-7 biosynthesis. Given this link, we targeted SinR to construct a dynamic regulatory system, aiming to modulate MK-7 production by leveraging SinR's regulatory influence.</p><p><strong>Results: </strong>A modular PhrC-RapC-SinR QS system is developed to dynamic regulate side pathway of MK-7. In this study, first, we analyzed the SinR-based gene expression regulation system in B. subtilis 168 (BS168). We constructed a promoter library of different abilities, selected suitable promoters from the library, and performed mutation screening on the selected promoters. Furthermore, we constructed a PhrC-RapC-SinR QS system to dynamically control the synthesis of Phe, Tyr, Trp, folic acid, dihydroxybenzoate, hydroxybutanone in MK-7 synthesis in BS168. Cell growth and efficient synthesis of the MK-7 production can be dynamically balanced by this QS system. Using this system to balance cell growth and product fermentation, the MK-7 yield was ultimately increased by 6.27-fold, from 13.95 mg/L to 87.52 mg/L.</p><p><strong>Conclusion: </strong>In summary, the PhrC-RapC-SinR QS system has been successfully integrated with biocatalytic functions to achieve dynamic metabolic pathway control in BS168, which has potential applicability to a large number of microorganisms to fine-tune gene expression and enhance the production of metabolites.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"88"},"PeriodicalIF":4.3,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12010548/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063832","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":"Linking a polyketide synthase gene cluster to 6-pentyl-alpha-pyrone, a Trichoderma metabolite with diverse bioactivities.","authors":"Daniel Flatschacher, Alexander Eschlböck, Siebe Pierson, Ulrike Schreiner, Valentina Stock, Arne Schiller, David Ruso, Maria Doppler, Veronika Ruzsanyi, Mario Gründlinger, Christoph Büschl, Rainer Schuhmacher, Susanne Zeilinger","doi":"10.1186/s12934-025-02718-9","DOIUrl":"https://doi.org/10.1186/s12934-025-02718-9","url":null,"abstract":"<p><strong>Background: </strong>Members of the fungal genus Trichoderma are well-known for their mycoparasitic and plant protecting activities, rendering them important biocontrol agents. One of the most significant specialized metabolites (SMs) produced by various Trichoderma species is the unsaturated lactone 6-pentyl-alpha-pyrone (6-PP). Although first identified more than 50 years ago and having pronounced antifungal and plant growth-promoting properties, the biosynthetic pathway of 6-PP still remains unresolved.</p><p><strong>Results: </strong>Here, we demonstrate that 6-PP is biosynthesized via the polyketide biosynthesis pathway. We identified Pks1, an iterative type I polyketide synthase, as crucial for its biosynthesis in Trichoderma atroviride, a species recognized for its prominent 6-PP production abilities. Phylogenetic and comparative genomic analyses revealed that the pks1 gene is part of a biosynthetic gene cluster conserved in those Trichoderma species that are known to produce 6-PP. Deletion of pks1 caused a complete loss of 6-PP production in T. atroviride and a significant reduction in antifungal activity against Botrytis cinerea and Rhizoctonia solani. Surprisingly, the absence of pks1 led to enhanced lateral root formation in Arabidopsis thaliana during interaction with T. atroviride. Transcriptomic analysis revealed co-regulation of pks1 with adjacent genes, including candidates coding for a C3H1-type zinc finger protein and lytic polysaccharide monooxygenase, suggesting coordination between 6-PP biosynthesis and environmental response mechanisms.</p><p><strong>Conclusion: </strong>Our findings establish pks1 as an essential gene for 6-PP biosynthesis in T. atroviride, providing novel insights into the production of one of the most significant compounds of this mycoparasite. These findings may pave the way for the development of improved biocontrol agents and the application of 6-PP as potent biopesticide contributing to an eco-friendly and sustainable way of plant disease management.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"89"},"PeriodicalIF":4.3,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12010586/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144021842","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":"Effects of Magnaporthe oryzae cell-free filtrate on the secondary metabolism of Streptomyces bikiniensis HD-087: a non-targeted metabolomics analysis.","authors":"Jiahan Gang, Qingqing Tian, Chunmei Du","doi":"10.1186/s12934-025-02711-2","DOIUrl":"https://doi.org/10.1186/s12934-025-02711-2","url":null,"abstract":"<p><p>Rice blast, a disease caused by Magnaporthe oryzae, significantly threatens global rice production. To improve the anti-M. oryzae activity of Streptomyces bikiniensis HD-087 metabolites, the effects of inducer, Magnaporthe oryzae acellular filtrate, on secondary metabolism of S. bikiniensis HD-087 were studied. The results showed that M. oryzae cell-free filtrate cultured for 96 h served as the most effective inducer, significantly enhancing the anti-M. oryzae activity of metabolites of S. bikiniensis HD-087 and increasing the diameter of the inhibitory zone by 2.96 mm. The inhibition rates of M. oryzae colony diameter and spore germination in the induced group were 12.39% and 39.6% higher than those in the non-induced group, respectively. Metabolomic profiling of strain HD-087 highlighted substantial differences between the induced and non-induced groups. At 48 h of fermentation, a total of 705 distinct metabolites were identified, while at 96 h this number decreased to 321. Moreover, induction markedly altered primary pathways such as the tricarboxylic acid cycle, amino acid biosynthesis, and fatty acid metabolism in S. bikiniensis HD-087. qPCR analysis showed that nrps genes and pks genes in the induced group were significantly up-regulated by 9.92 ± 0.51 and 2.71 ± 0.17 times, respectively, and biotin carboxylase activity was also increased 26.63%. These results provide a theoretical basis for using inducers to enhance the antimicrobial ability of Streptomyces.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"90"},"PeriodicalIF":4.3,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12013144/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144007803","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":"Development and construction of a novel Bacillus subtilis autoinducible extracellular expression system based on a LuxI/R device.","authors":"Bin Wang, Keyi Wang, Xiuyue Zhao, Zemin Fang, Yanyan Zhao, Yulu Fang, Yazhong Xiao, Dongbang Yao","doi":"10.1186/s12934-025-02719-8","DOIUrl":"https://doi.org/10.1186/s12934-025-02719-8","url":null,"abstract":"<p><strong>Background: </strong>Microbial chassis expression systems are valuable tools in biotechnology and synthetic biology, and Bacillus subtilis is an important industrial microbial chassis. Quorum sensing (QS)-based dynamic regulation is widely used to automatically activate gene expression in response to changes in cell density. The main bottleneck currently limiting the use of exogenous QS systems in B. subtilis for efficient autoinducible extracellular expression of recombinant proteins is their low level of autoinducible expression.</p><p><strong>Results: </strong>A novel B. subtilis autoinducible extracellular expression system based on the LuxI/R-type QS system (lux system) of Vibrio fischeri was developed in which the autoinducible expression of the lux system was enhanced by engineering the sensing module and response module promoters. By engineering the sensing module promoter SP_luxI core region (- 10 and - 35 elements) and critical region (UP and spacer elements), and the response module promoter RP_luxIR6 core region and lux box copy number in the original LuxI/R device (S0-R0), the high-expression Sc-R2 construct was obtained. After shake flask and 3-L fermenter fermentation, the extracellular amylase activity obtained with Sc-R2 was 2.7- and 3.1-fold greater, respectively, than that obtained with the well-characterized promoter P_veg. Sc-R2 achieved 2.6-fold greater extracellular activity than S0-R0 when either levansucrase or invertase was used as a reporter protein. Overall, the B. subtilis autoinducible extracellular expression system developed in this study showed good generalizability and application potential for industrial-scale fermentation.</p><p><strong>Conclusions: </strong>To our knowledge, this is the first study to report enhanced autoinducible expression of the lux system in B. subtilis by engineering the sensing module promoter SP_luxI sequence and the lux box copy number of the response module promoter RP_luxIR6. This study further expands the application potential of the B. subtilis expression system in synthetic biology.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"86"},"PeriodicalIF":4.3,"publicationDate":"2025-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008979/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144022855","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":"Optimization of thermostable amylolytic enzyme production from Bacillus cereus isolated from a recreational warm spring via Box Behnken design and response surface methodology.","authors":"Oluwaseun Abosede Adetiloye, Bamidele Ogbe Solomon, Japhael Abel Omolaiye, Eriola Betiku","doi":"10.1186/s12934-025-02709-w","DOIUrl":"https://doi.org/10.1186/s12934-025-02709-w","url":null,"abstract":"<p><p>This study aimed to find a source for local amylase-producing microbes. Sixteen isolates were obtained from the water samples from the warm spring and characterized based on morphological and biochemical tests. The 16S rRNA molecular identification technique confirmed the most potent isolate as Bacillus cereus. The thermophilic property of the bacterium demonstrated that it could withstand temperatures of up to 80 °C. One-factor-at-a-time (OFAT) and Box Behnken Design (BBD) coupled with response surface methodology (RSM) optimization techniques were used to improve amylase production. OFAT established optimal physical parameter conditions as the starch concentration of 5% w/v, inoculum volume of 2% v/v, pH of 8, incubation temperature of 45 °C, and 48 h of incubation, leading to amylase activity of 172.6 U/mL by the isolated B. cereus. A quadratic mathematical model with a coefficient of determination (R²) of 0.9957 was established for the amylase production process. Enhanced amylase activity of 196.02 U/mL was achieved with BBD-RSM under optimal growth conditions of pH of 7, incubation time of 48 h, substrate concentration of 5% w/v of starch, and at 45 °C, a 1.2-fold increase compared to the OFAT method. The B. cereus strain isolated from the warm spring was a mildly thermophilic bacterium with the potential for synthesizing amylolytic enzymes with characteristics beneficial for commercial utilization.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"87"},"PeriodicalIF":4.3,"publicationDate":"2025-04-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12008982/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144029366","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":"Biodegradation of p-nitrophenol by Rhodococcus sp. 21391 unveils a two-component p-nitrophenol monooxygenase with broad substrate specificity.","authors":"Jian Yang, Shanshan Lin, Wei Li, Xianjie Wang, Ru Li","doi":"10.1186/s12934-025-02712-1","DOIUrl":"https://doi.org/10.1186/s12934-025-02712-1","url":null,"abstract":"<p><strong>Background: </strong>Bioremediation relying on highly efficient degrading bacteria constitutes a promising and sustainable avenue for controlling and reducing nitrophenol contamination in the environment. A thorough understanding of the bacterial degradation mechanism of nitrophenol is of paramount importance for supporting the development of efficient microbial remediation technology.</p><p><strong>Results: </strong>In this study, a new bacterium, Rhodococcus sp. 21391, endowed with superior p-nitrophenol (PNP) degradation ability was obtained. Genomic and comparative proteomic analyses revealed that it utilizes the 1,2,4-benzenetriol (BT) pathway for PNP degradation. The catalytic properties of the two-component p-nitrophenol monooxygenase RsNcpAB from the strain were investigated in vitro. The enzyme exhibited a broad substrate selectivity, catalyzing the oxidation of various nitrophenols and halogenated phenols, with significant potential for further research and development. Additionally, the crystal structure of the oxidative component of p-nitrophenol monooxygenase, RsNcpA, was determined. Structural analysis and site-directed mutagenesis revealed that residues Arg100 and His293 in the active site play a crucial role in enzyme catalysis, and a catalytic mechanism model was subsequently proposed.</p><p><strong>Conclusions: </strong>This study reports a high-performance nitrophenol-degrading bacterium and enzyme, and reveals their mechanisms at the molecular level. These findings increase the understanding of the bacterial degradation of nitrophenol, thereby providing a crucial foundation for the development of efficient bioremediation technologies.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"85"},"PeriodicalIF":4.3,"publicationDate":"2025-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12007285/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144026665","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 IgG-Fc-Fused multiepitope vaccine against Brucella: robust immunogenicity.","authors":"Aodi Wu, Yuting Zhang, Caidong Liu, Kaiat Zhumanov, Tao He, Kexin Yan, Honghuan Li, Shuangshaung Fu, Xin Li, Wenxiang Zhang, Chuang Meng, Changsuo Zhang, Jinliang Sheng, Zhongchen Ma, Mingguo Xu, Junbo Zhang, Jihai Yi, Yueli Wang","doi":"10.1186/s12934-025-02713-0","DOIUrl":"https://doi.org/10.1186/s12934-025-02713-0","url":null,"abstract":"<p><p>Brucellosis is one of the most common zoonotic diseases caused by Brucella spp. However, there is currently no Brucella vaccine available for humans. Although some attenuated live vaccines have been approved for animals, their protective efficacy is suboptimal. In previous studies, we utilized an epitope- and structure-based vaccinology platform to identify the immunodominant epitopes of Brucella antigens OMP19, OMP16, OMP25, and L7/L12, and constructed the multi-epitope vaccine MEV-Fc against Brucella. In this study, OMP19, OMP16, OMP25, and L7/L12, and MEV-Fc was expressed and purified via an Escherichia coli expression system, which validated that MEV-Fc possesses high immunological efficacy and exerts a significant protective effect in BALB/c mice within the Brucella infection model. MEV-Fc enhanced Th1 and Th2 immune responses and strongly induced the production of the pro-inflammatory cytokine IFN-γ. Furthermore, MEV-Fc protected mice against Brucella infection compared to control group (PBS). In conclusion, our results provide new insights and data support for the development of human Brucella vaccines.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"84"},"PeriodicalIF":4.3,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11998165/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001595","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":"Improved methyl supply for 5-methyltetrahydrofolate production in E. coli through a novel C1 transfer pathway.","authors":"Wen Liu, Jing Guo, Wei Lu, Tao Cheng, Yiting Li, Mo Xian, Rubing Zhang","doi":"10.1186/s12934-025-02707-y","DOIUrl":"https://doi.org/10.1186/s12934-025-02707-y","url":null,"abstract":"<p><strong>Background: </strong>L-5-Methyltetrahydrofolate (5-MTHF) is the sole biologically active form of folate present in human blood and serves as an essential nutritional supplement. While microbial biosynthesis of 5-MTHF offers a sustainable alternative to chemical synthesis, its low yield limits industrial potential.</p><p><strong>Results: </strong>In this study, strategies for improving the methyl supply combined with engineering the tetrahydrofolate (THF) synthetic pathway were employed in E. coli to increase 5-MTHF production. First, a new exogenous C1 pathway was introduced to improve the intracellular methyl supply through acetyl-CoA breakdown. High expression of key rate-limiting genes folE, folP and purU enhanced metabolic flux of THF pathway, resulting in a 5-MTHF titer of 1.075 mg/L during shake-flask fermentation. A subsequent increase in 5-MTHF production was achieved by knocking out the metE gene, which is involved in the consumption of 5-MTHF. The best engineered strain, M3012, produced 8.2 mg/L 5-MTHF in a 5 L bioreactor via fed-batch fermentation, which presented the highest 5-MTHF titer to date.</p><p><strong>Conclusion: </strong>We successfully engineered E. coli by introducing a novel exogenous C1 metabolic pathway to augment the methyl donor pool essential for the biosynthesis of 5-MTHF. Further metabolic optimizations, including the enhancement of the THF precursor flux and the elimination of competing degradation pathways, developed a recombinant strain with significantly increased yield, which paves the way for industrial production of 5-MTHF.</p>","PeriodicalId":18582,"journal":{"name":"Microbial Cell Factories","volume":"24 1","pages":"83"},"PeriodicalIF":4.3,"publicationDate":"2025-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11998260/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144033579","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}