Zhao Weijun, Shi Feng, Hang Baojian, Huang Lei, Cai Jin, Xu Zhinan
Key Laboratory of Biomass Chemical Engineering (Ministry of Education), College of Chemical and Biological Engineering, Zhejiang University, 38 Zheda Road, Hangzhou, 310027, China.
Shandong Institute for Food and Drug Control, Jinan, 250101, China.
Appl Biochem Biotechnol. 2016 Mar;178(6):1263-72. doi: 10.1007/s12010-015-1943-1. Epub 2016 Jan 4.
S-Adenosyl-L-methionine (SAM) plays important roles in trans-methylation, trans-sulfuration, and polyamine synthesis in all living cells, and it is also an effective cure for liver disease, depressive syndromes, and osteoarthritis. The increased demands of SAM in pharmaceuticals industry have aroused lots of attempts to improve its production. In this study, a multiple-copy integrative plasmid pYMIKP-SAM2 was introduced into the chromosome of wild-type Saccharomyces cerevisiae strain ZJU001 to construct the recombined strain R1-ZJU001. Further studies showed that the recombinant yeast exhibited higher enzymatic activity of methionine adenosyltransferase and improved its SAM biosynthesis. With a three-phase fed-batch strategy in 15-liter bench-top fermentor, 8.81 g/L SAM was achieved after 52 h cultivation of R1-ZJU001, about 27.1 % increase over its parent strain ZJU001, whereas the SAM content was also improved from 64.6 mg/g DCW to 91.0 mg/g DCW. Our results shall provide insights into the metabolic engineering of SAM pathway in yeast for improved productivity of SAM and subsequent industrial applications.
S-腺苷-L-甲硫氨酸(SAM)在所有活细胞的转甲基作用、转硫作用和多胺合成中发挥着重要作用,并且它还是治疗肝病、抑郁综合征和骨关节炎的有效药物。制药行业对SAM需求的增加引发了许多提高其产量的尝试。在本研究中,将多拷贝整合质粒pYMIKP-SAM2导入野生型酿酒酵母菌株ZJU001的染色体中,构建重组菌株R1-ZJU001。进一步研究表明,重组酵母表现出更高的甲硫氨酸腺苷转移酶活性,并改善了其SAM生物合成。在15升台式发酵罐中采用三相补料分批策略,R1-ZJU001培养52小时后获得了8.81 g/L的SAM产量,比其亲本菌株ZJU001提高了约27.1%,同时SAM含量也从64.6 mg/g干细胞重量提高到91.0 mg/g干细胞重量。我们的结果将为酵母中SAM途径的代谢工程提供见解,以提高SAM的产量并用于后续工业应用。