System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production
Abstract S-adenosyl-L-methionine (SAM) is an important compound with significant pharmaceutical and nutraceutical applications. Currently, microbial fermentation is dominant in SAM production, which remains challenging due to its complex biosynthetic pathway and insufficient precursor availability....
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
SpringerOpen
2025-03-01
|
| Series: | Bioresources and Bioprocessing |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40643-025-00858-9 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850039507739475968 |
|---|---|
| author | Liangzhuang Tan Yuehan Zhang Ping Liu Yihang Wu Zuoyu Huang Zhongce Hu Zhiqiang Liu Yuanshan Wang Yuguo Zheng |
| author_facet | Liangzhuang Tan Yuehan Zhang Ping Liu Yihang Wu Zuoyu Huang Zhongce Hu Zhiqiang Liu Yuanshan Wang Yuguo Zheng |
| author_sort | Liangzhuang Tan |
| collection | DOAJ |
| description | Abstract S-adenosyl-L-methionine (SAM) is an important compound with significant pharmaceutical and nutraceutical applications. Currently, microbial fermentation is dominant in SAM production, which remains challenging due to its complex biosynthetic pathway and insufficient precursor availability. In this study, a multimodule engineering strategy based on CRISPR/Cas9 was established to improve the SAM productivity of Saccharomyces cerevisiae. This strategy consists of (1) improving the growth of S. cerevisiae by overexpressing the hxk2 gene; (2) enhancing the metabolic flux toward SAM synthesis by upregulating the expression of the aat1, met17, and sam2 genes and weakening the synthesis pathway of L-threonine; (3) elevating precursor ATP synthesis by introducing the vgb gene; (4) blocking the SAM degradation pathway by knocking out the sah1 and spe2 genes. The SAM titer of the resulting mutant AU18 reached 1.87 g/L, representing an increase of 227.67% compared to the parental strain. With optimal medium, SAM titer of mutant AU18 reached 2.46 g/L in flask shake fermentation. The SAM titer of mutant AU18 further reached 13.96 g/L after 96 h incubation with a continuous L-Met feeding strategy in a 5 L fermenter. Therefore, with comprehensive optimization of both synthesis and degradation pathways of SAM, a multimodule strategy was established, which significantly elevated the SAM production of S. cerevisiae. This laid a foundation for the construction of hyperproducer for SAM and other valuable amino acids or chemicals. |
| format | Article |
| id | doaj-art-7fc02ee8d7dc41a6af5afd1595391121 |
| institution | DOAJ |
| issn | 2197-4365 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Bioresources and Bioprocessing |
| spelling | doaj-art-7fc02ee8d7dc41a6af5afd15953911212025-08-20T02:56:20ZengSpringerOpenBioresources and Bioprocessing2197-43652025-03-0112111510.1186/s40643-025-00858-9System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM productionLiangzhuang Tan0Yuehan Zhang1Ping Liu2Yihang Wu3Zuoyu Huang4Zhongce Hu5Zhiqiang Liu6Yuanshan Wang7Yuguo Zheng8Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyKey Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of TechnologyAbstract S-adenosyl-L-methionine (SAM) is an important compound with significant pharmaceutical and nutraceutical applications. Currently, microbial fermentation is dominant in SAM production, which remains challenging due to its complex biosynthetic pathway and insufficient precursor availability. In this study, a multimodule engineering strategy based on CRISPR/Cas9 was established to improve the SAM productivity of Saccharomyces cerevisiae. This strategy consists of (1) improving the growth of S. cerevisiae by overexpressing the hxk2 gene; (2) enhancing the metabolic flux toward SAM synthesis by upregulating the expression of the aat1, met17, and sam2 genes and weakening the synthesis pathway of L-threonine; (3) elevating precursor ATP synthesis by introducing the vgb gene; (4) blocking the SAM degradation pathway by knocking out the sah1 and spe2 genes. The SAM titer of the resulting mutant AU18 reached 1.87 g/L, representing an increase of 227.67% compared to the parental strain. With optimal medium, SAM titer of mutant AU18 reached 2.46 g/L in flask shake fermentation. The SAM titer of mutant AU18 further reached 13.96 g/L after 96 h incubation with a continuous L-Met feeding strategy in a 5 L fermenter. Therefore, with comprehensive optimization of both synthesis and degradation pathways of SAM, a multimodule strategy was established, which significantly elevated the SAM production of S. cerevisiae. This laid a foundation for the construction of hyperproducer for SAM and other valuable amino acids or chemicals.https://doi.org/10.1186/s40643-025-00858-9S-adenosyl-L-MethionineSaccharomyces cerevisiaeCRISPR/Cas9 systemCombinatorial metabolic engineeringFed-batch fermentationTCA cycle |
| spellingShingle | Liangzhuang Tan Yuehan Zhang Ping Liu Yihang Wu Zuoyu Huang Zhongce Hu Zhiqiang Liu Yuanshan Wang Yuguo Zheng System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production Bioresources and Bioprocessing S-adenosyl-L-Methionine Saccharomyces cerevisiae CRISPR/Cas9 system Combinatorial metabolic engineering Fed-batch fermentation TCA cycle |
| title | System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production |
| title_full | System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production |
| title_fullStr | System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production |
| title_full_unstemmed | System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production |
| title_short | System metabolic engineering modification of Saccharomyces cerevisiae to increase SAM production |
| title_sort | system metabolic engineering modification of saccharomyces cerevisiae to increase sam production |
| topic | S-adenosyl-L-Methionine Saccharomyces cerevisiae CRISPR/Cas9 system Combinatorial metabolic engineering Fed-batch fermentation TCA cycle |
| url | https://doi.org/10.1186/s40643-025-00858-9 |
| work_keys_str_mv | AT liangzhuangtan systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT yuehanzhang systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT pingliu systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT yihangwu systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT zuoyuhuang systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT zhongcehu systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT zhiqiangliu systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT yuanshanwang systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction AT yuguozheng systemmetabolicengineeringmodificationofsaccharomycescerevisiaetoincreasesamproduction |