| 利用重组Pichia pastoris生产腺苷甲硫氨酸 |
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| 引用本文： | 李东阳,于健,田露,吉鑫松,袁中一.利用重组Pichia pastoris生产腺苷甲硫氨酸[J].生物工程学报,2002,18(3):295-299. |
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| 作者姓名： | 李东阳 于健 田露 吉鑫松 袁中一 |
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| 作者单位： | 中国科学院上海生命科学院生物化学与细胞生物学研究所,上海,200031 |
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| 摘 要： | 为改造甲醇利用型酵母Pichia pastoris来生产腺苷甲硫氨酸（SAM，S－adenosyl-L-methionine)，我们将一个带有SAM合成酶基因的胞内表达质粒转化入Pichia pastoris菌株GS115，经过G418抗性筛选得到一株有两个基因拷贝的转化子。该菌在含有甲醇和甲硫氨酸的培养基中生长5d后，其细胞内的SAM的产量比原始菌株提高了30余倍。对该菌生产SAM的培养基中的碳源与氮源进行了优化，结果显示碳源的控制对该菌SAM产量的影响很大。在试管水平，该菌在含有0．75％的L-methionine并且碳源和有机氮源经过一定程度优化的培养基中，生长6d后SAM产量达到1．58g/L。
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| 关 键 词： | 腺苷甲硫氨酸高产菌 重组 SAM合成酶 |
| 文章编号： | 1000-3061(2002)03-0295-05 |
| 修稿时间： | 2001年11月23 |
Production of SAM by Recombinant Pichia pastoris |
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| LI Dong-Yang,YU Jian,TIAN Lu,JI Xin-Song,YUAN Zhong-Yi.Production of SAM by Recombinant Pichia pastoris[J].Chinese Journal of Biotechnology,2002,18(3):295-299. |
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| Authors: | LI Dong-Yang YU Jian TIAN Lu JI Xin-Song YUAN Zhong-Yi |
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| Affiliation: | Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. |
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| Abstract: | To utilize Pichia pastoris to produce S-adenosyl-L-methionine (SAM), an intracellular expression vector harboring S. cerevisiae SAM2 was transformed into GS115. A recombinant strain having 2 copies of expression cassette was obtained through G418 resistance screening. This strain had higher SAM synthetase activity and higher SAM production capacity than the original strain, when cultured in medium containing methanol and methionine. The carbon source and nitrogen source of medium was optimized. The results showed SAM production by this strain was closely related to carbon metabolism. With supplementation of 0.2% glycerol every day from the beginning of 3rd day, this strain produced 1.58g/L SAM when cultured in a medium containing 0.75% L-methionine and optimized carbon and nitrogen source after 6 days. |
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| Keywords: | S-adenosyl-L-methionine high production strain recombinant SAM synthetase Pichia pastoris |
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