A Novel meso-Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for D-Amino Acid Synthesis

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	A Novel meso-Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for D-Amino Acid Synthesis
	Gao, Xiuzhen 1; Chen, Xi 1; Liu, Weidong 1; Feng, Jinhui 1; Wu, Qiaqing 1; Hua, Ling 2; Zhu, Dunming 1
刊名	APPLIED AND ENVIRONMENTAL MICROBIOLOGY
	2012-12-01
卷号	78 期号:24 页码:8595-8600
英文摘要	meso-Diaminopimelate dehydrogenase (meso-DAPDH) is an NADP(+)-dependent enzyme which catalyzes the reversible oxidative deamination on the D-configuration of meso-2,6-diaminopimelate to produce L-2-amino-6-oxopimelate. In this study, the gene encoding a meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum was cloned and expressed in Escherichia coli. In addition to the native substrate meso-2,6-diaminopimelate, the purified enzyme also showed activity toward D-alanine, D-valine, and D-lysine. This enzyme catalyzed the reductive amination of 2-keto acids such as pyruvic acid to generate D-amino acids in up to 99% conversion and 99% enantiomeric excess. Since meso-diaminopimelate dehydrogenases are known to be specific to meso-2,6-diaminopimelate, this is a unique wild-type meso-diaminopimelate dehydrogenase with a more relaxed substrate specificity and potential for D-amino acid synthesis. The enzyme is the most stable meso-diaminopimelate dehydrogenase reported to now. Two amino acid residues (F146 and M152) in the substrate binding sites of S. thermophilum meso-DAPDH different from the sequences of other known meso-DAPDHs were replaced with the conserved amino acids in other meso-DAPDHs, and assay of wild-type and mutant enzyme activities revealed that F146 and M152 are not critical in determining the enzyme's substrate specificity. The high thermostability and relaxed substrate profile of S. thermophilum meso-DAPDH warrant it as an excellent starting enzyme for creating effective D-amino acid dehydrogenases by protein engineering.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology ; Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology ; Microbiology
关键词[WOS]	CORYNEBACTERIUM-GLUTAMICUM ; BACILLUS-SPHAERICUS ; GENE ; SEQUENCE ; CREATION
收录类别	SCI
语种	英语
WOS记录号	WOS:000311213200012
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1324]
专题	天津工业生物技术研究所_生物催化与绿色化工朱敦明_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Natl Engn Lab Ind Enzymes, Tianjin, Peoples R China 2.DuPont Ind Biosci, China Res Ctr, Shanghai, Peoples R China
推荐引用方式 GB/T 7714	Gao, Xiuzhen,Chen, Xi,Liu, Weidong,et al. A Novel meso-Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for D-Amino Acid Synthesis[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY,2012,78(24):8595-8600.
APA	Gao, Xiuzhen.,Chen, Xi.,Liu, Weidong.,Feng, Jinhui.,Wu, Qiaqing.,...&Zhu, Dunming.(2012).A Novel meso-Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for D-Amino Acid Synthesis.APPLIED AND ENVIRONMENTAL MICROBIOLOGY,78(24),8595-8600.
MLA	Gao, Xiuzhen,et al."A Novel meso-Diaminopimelate Dehydrogenase from Symbiobacterium thermophilum: Overexpression, Characterization, and Potential for D-Amino Acid Synthesis".APPLIED AND ENVIRONMENTAL MICROBIOLOGY 78.24(2012):8595-8600.