Expression of secreted His-tagged S-adenosylmethionine synthetase in the methylotrophic yeast Pichia pastoris and its characterization, one-step purification, and immobilization

	Expression of secreted His-tagged S-adenosylmethionine synthetase in the methylotrophic yeast Pichia pastoris and its characterization, one-step purification, and immobilization
	Luo, YX; Yuan, ZY; Luo, GM; Zhao, FK
刊名	BIOTECHNOLOGY PROGRESS
	2008
卷号	24 期号:1 页码:214-220
通讯作者	Zhao, FK (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, Key Lab Proteom, Shanghai 200031, Peoples R China.,fkzhao@sibs.ac.cn
英文摘要	S-Adenosylmethionine synthetase (SAM synthetase) catalyzes the synthesis of S-adenosylmethionine (SAM), which plays an important role in cellular functions such as methylation, sulfuration, and polyamine synthesis. To develop a simple and effective way to enzymatically synthesize and produce SAM, a soluble form of SAM synthetase encoded by SAM2 from Saccharomyces cerevisiae was successfully produced at high level (similar to 200 mg/L) by the recombinant methylotrophic yeast Pichia pastoris. The secreted HiS(6)-tagged SAM synthetase was purified in a single chromatography step with a yield of approximately 82% for the total activity. The specific activity of the purified synthetase was 23.84 U/mg. The recombinant SAM synthetase could be a kind of allosteric enzyme with negative regulation. The enzyme functioned optimally at a temperature of 35 degrees C and pH 8.5. The stability of the recombinant synthetase and the effectiveness of different factors in preventing the enzyme from inactivation were also studied. Additional experiments were performed in which the recombinant SAM synthetase was purified and immobilized in one step using immobilized metal-chelate affinity chromatography. The immobilized synthetase was found to be 40.4% of the free enzyme activity in catalyzing the synthesis of SAM from DL-Met and ATP.
学科主题	Biotechnology & Applied Microbiology; Food Science & Technology
类目[WOS]	Biotechnology & Applied Microbiology ; Food Science & Technology
关键词[WOS]	ADENOSYL-L-METHIONINE ; SACCHAROMYCES-CEREVISIAE ; ENZYMATIC-SYNTHESIS ; ESCHERICHIA-COLI ; BAKERS-YEAST ; RAT-LIVER ; ADENOSYLTRANSFERASE ; TRANSMETHYLATION ; ACTIVATION ; ENZYMES
收录类别	SCI
语种	英语
WOS记录号	WOS:000252878700030
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1325]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Luo, YX,Yuan, ZY,Luo, GM,et al. Expression of secreted His-tagged S-adenosylmethionine synthetase in the methylotrophic yeast Pichia pastoris and its characterization, one-step purification, and immobilization[J]. BIOTECHNOLOGY PROGRESS,2008,24(1):214-220.
APA	Luo, YX,Yuan, ZY,Luo, GM,&Zhao, FK.(2008).Expression of secreted His-tagged S-adenosylmethionine synthetase in the methylotrophic yeast Pichia pastoris and its characterization, one-step purification, and immobilization.BIOTECHNOLOGY PROGRESS,24(1),214-220.
MLA	Luo, YX,et al."Expression of secreted His-tagged S-adenosylmethionine synthetase in the methylotrophic yeast Pichia pastoris and its characterization, one-step purification, and immobilization".BIOTECHNOLOGY PROGRESS 24.1(2008):214-220.