TY - JOUR
T1 - Identification of novel metabolic engineering targets for S-adenosyl-L-methionine production in Saccharomyces cerevisiae via genome-scale engineering
AU - Dong, Chang
AU - Schultz, J. Carl
AU - Liu, Wei
AU - Lian, Jiazhang
AU - Huang, Lei
AU - Xu, Zhinan
AU - Zhao, Huimin
N1 - Funding Information:
This work was supported by the Natural Science Foundation of Zhejiang Province ( LR20B060003 to JL), the Natural Science Foundation of China ( 21808199 to JL and 21576232 to ZX), the National Key Research and Development Program of China ( 2018YFA0901800 to JL), and the U.S. Department of Energy ( DE-SC0018260 and DE-SC0018420 to HZ). We would like to thank Prof. Takashi Ito from the University of Tokyo for sharing the SAM biosensor plasmid. From the Roy J. Carver Biotechnology Center at the University of Illinois at Urbana-Champaign, we would like to thank Dr. Barbara Pilas for assistance with FACS, Dr. Lucas Li for assistance with metabolomics, and Dr. Chris Wright and Dr. Alvaro Hernandez for performing RNA-sequencing library preparation and next-generation sequencing.
Funding Information:
This work was supported by the Natural Science Foundation of Zhejiang Province (LR20B060003 to JL), the Natural Science Foundation of China (21808199 to JL and 21576232 to ZX), the National Key Research and Development Program of China (2018YFA0901800 to JL), and the U.S. Department of Energy (DE-SC0018260 and DE-SC0018420 to HZ). We would like to thank Prof. Takashi Ito from the University of Tokyo for sharing the SAM biosensor plasmid. From the Roy J. Carver Biotechnology Center at the University of Illinois at Urbana-Champaign, we would like to thank Dr. Barbara Pilas for assistance with FACS, Dr. Lucas Li for assistance with metabolomics, and Dr. Chris Wright and Dr. Alvaro Hernandez for performing RNA-sequencing library preparation and next-generation sequencing.
Publisher Copyright:
© 2021 International Metabolic Engineering Society
PY - 2021/7
Y1 - 2021/7
N2 - S-Adenosyl-L-methionine (SAM) is an important intracellular metabolite and widely used for treatment of various diseases. Although high level production of SAM had been achieved in yeast, novel metabolic engineering strategies are needed to further enhance SAM production for industrial applications. Here genome-scale engineering (GSE) was performed to identify new targets for SAM overproduction using the multi-functional genome-wide CRISPR (MAGIC) system, and the effects of these newly identified targets were further validated in industrial yeast strains. After 3 rounds of FACS screening and characterization, numerous novel targets for enhancing SAM production were identified. In addition, transcriptomic and metabolomic analyses were performed to investigate the molecular mechanisms for enhanced SAM accumulation. The best combination (upregulation of SNZ3, RFC4, and RPS18B) improved SAM productivity by 2.2-fold and 1.6-fold in laboratory and industrial yeast strains, respectively. Using GSE of laboratory yeast strains to guide industrial yeast strain engineering presents an effective approach to design microbial cell factories for industrial applications.
AB - S-Adenosyl-L-methionine (SAM) is an important intracellular metabolite and widely used for treatment of various diseases. Although high level production of SAM had been achieved in yeast, novel metabolic engineering strategies are needed to further enhance SAM production for industrial applications. Here genome-scale engineering (GSE) was performed to identify new targets for SAM overproduction using the multi-functional genome-wide CRISPR (MAGIC) system, and the effects of these newly identified targets were further validated in industrial yeast strains. After 3 rounds of FACS screening and characterization, numerous novel targets for enhancing SAM production were identified. In addition, transcriptomic and metabolomic analyses were performed to investigate the molecular mechanisms for enhanced SAM accumulation. The best combination (upregulation of SNZ3, RFC4, and RPS18B) improved SAM productivity by 2.2-fold and 1.6-fold in laboratory and industrial yeast strains, respectively. Using GSE of laboratory yeast strains to guide industrial yeast strain engineering presents an effective approach to design microbial cell factories for industrial applications.
KW - CRISPR
KW - Genome-scale engineering
KW - Industrial S. cerevisiae
KW - S-Adenosyl-L-methionine
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U2 - 10.1016/j.ymben.2021.03.005
DO - 10.1016/j.ymben.2021.03.005
M3 - Article
C2 - 33713797
AN - SCOPUS:85106585651
VL - 66
SP - 319
EP - 327
JO - Metabolic Engineering
JF - Metabolic Engineering
SN - 1096-7176
ER -