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Systematically redesigning and optimizing the expression of (D)-lactate dehydrogenase efficiently produces high-optical-purity (D)-lactic acid in Saccharomyces cerevisiae

文献类型：期刊论文

作者Zhong, Wei1,2; Yang, Maohua1; Mu, Tingzhen1; Wu, Fan1,3; Hao, Xuemi1,2; Chen, Ruonan4; Sharshar, Moustafa Mohamed1,2; Thygesen, Anders5; Wang, Qinhong6; Xing, Jianmin1
刊名BIOCHEMICAL ENGINEERING JOURNAL
出版日期2019-04-15
卷号144页码:217-226
ISSN号1369-703X
关键词Synthetic biology (D)-lactic acid Double enzyme-coupled system Chromosomal integration Saccharomyces cerevisiae
DOI10.1016/j.bej.2018.09.013
英文摘要(D)-lactic acid ((D)-LA) is gaining increased attention as it can improve the thermostability of poly lactic acid. Acid tolerant Saccharomyces cerevisiae is a good host for (D)-LA fermentation. High catalytic efficiency of (D)-lactate dehydrogenase ((D)-LDH, EC 1.1.1.28) is crucial for the production of (D)-LA in yeast. Here, a synthetic biology approach was used to construct high-producing (D)-LA strains by redesigning and optimization of (D)-LDH expression by a combination of different promoters, terminators and (D)-LDHs. The pyruvate decarboxylase-deficient mutant strain TAMH was used as host strain for optimizing the 40 (D)-LDH expression cassettes. The TCSt strain harboring the pTCSt plasmid with the TEF1 promoter, E. coli (D)-LDH and Synth25 synthetic short terminator produced 5.8 g/L (D)-LA with an optical purity of 99.9%. The production of (D)-LA was further improved by integrating this high expression cassette into the Ty1 transposable element of the YIP-01 strain with deleted Pdc1 and Pdc6. The resulting strain YIP-pTCSt-301(CGMCC2.5726) was screened by a double enzyme-coupled system. Genomes sequencing of the strain revealed three copies of the (D)-LDH expression cassette. This strain was further improved by deleting the Jen1, Cyb2, Dld1, and Adh1 genes and the resulting strain YIP-J-C-D-A1 (CGMCC2.5783) produced 80.0 g/L (D)-LA with a yield of 0.6 g/g glucose and a volumetric productivity of 1.1 g/L/h in fed-batch fermentation under non-neutralization conditions.
WOS关键词D-LACTIC ACID ; D-LACTATE DEHYDROGENASE ; ESCHERICHIA-COLI ; YEAST ; GENE ; FERMENTATION ; PATHWAY ; DNA ; OVEREXPRESSION ; PURIFICATION
资助项目Industrial Biotechnology Program of Tianjin Municipal Science and Technology Commission[14ZCZDSY00066] ; Science and Technology Service Network Initiative of Chinese academy of Sciences[KFJ-SW-STS-165]
WOS研究方向Biotechnology & Applied Microbiology ; Engineering
语种英语
出版者ELSEVIER SCIENCE BV
WOS记录号WOS:000462419600025
资助机构Industrial Biotechnology Program of Tianjin Municipal Science and Technology Commission ; Science and Technology Service Network Initiative of Chinese academy of Sciences
源URL[http://ir.ipe.ac.cn/handle/122111/28239]  
专题中国科学院过程工程研究所
通讯作者Wang, Qinhong; Xing, Jianmin
作者单位1.Chinese Acad Sci, Key Lab Green Proc & Engn, State Key Lab Biochem Engn, Inst Proc Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Univ Calif San Diego, Div Biol Sci, San Diego, CA 92092 USA
4.Beijing Univ Chem Technol, Beijing 100029, Peoples R China
5.Tech Univ Denmark, Prot Chem & Enzyme Technol Div, Dept Biochem & Biomed, DK-2800 Lyngby, Denmark
6.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Key Lab Syst Microbial Biotechnol, Tianjin 300308, Peoples R China
推荐引用方式
GB/T 7714		 Zhong, Wei,Yang, Maohua,Mu, Tingzhen,et al. Systematically redesigning and optimizing the expression of (D)-lactate dehydrogenase efficiently produces high-optical-purity (D)-lactic acid in Saccharomyces cerevisiae[J]. BIOCHEMICAL ENGINEERING JOURNAL,2019,144:217-226.
APA Zhong, Wei.,Yang, Maohua.,Mu, Tingzhen.,Wu, Fan.,Hao, Xuemi.,...&Xing, Jianmin.(2019).Systematically redesigning and optimizing the expression of (D)-lactate dehydrogenase efficiently produces high-optical-purity (D)-lactic acid in Saccharomyces cerevisiae.BIOCHEMICAL ENGINEERING JOURNAL,144,217-226.
MLA Zhong, Wei,et al."Systematically redesigning and optimizing the expression of (D)-lactate dehydrogenase efficiently produces high-optical-purity (D)-lactic acid in Saccharomyces cerevisiae".BIOCHEMICAL ENGINEERING JOURNAL 144(2019):217-226.

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来源：过程工程研究所

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