Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma

doi:10.3390/md16030080

CORC > 深海科学与工程研究所 > 中国科学院深海科学与工程研究所 > 深海科学研究部 > 深海生物学研究室 > 深海生物蛋白质组学及分子生物学研究组

	Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma
	Li, Yun-liang 1; Han, Zhuang4 ; Chen, Xi 2; Chen, Qi-jia 2; Wang, Yong4 ; He, Li-sheng4 ; He, Li-sheng(Chinese Acad Sci, Inst Deep Sea Sci & Engn, Dept Life Sci, Sanya 572000, Peoples R China); Wang, Shao-lu3,4
刊名	MARINE DRUGS
	2018-03-01
卷号	16 期号:3
关键词	N-acetylneuraminate Lyase Symbiotic Microbe N-acetylneuraminic Acid Mycoplasma
DOI	10.3390/md16030080
产权排序	第1完成单位 ; 第2完成单位 ; 第3完成单位 ; 第4完成单位
文献子类	Article
英文摘要	N-acetylneuraminic acid (Neu5Ac) based novel pharmaceutical agents and diagnostic reagents are highly required in medical fields. However, N-acetylneuraminate lyase (NAL) for Neu5Ac synthesis is not applicable for industry due to its low catalytic efficiency. In this study, we biochemically characterized a deep-sea NAL enzyme (abbreviated form: MyNal) from a symbiotic Mycoplasma inhabiting the stomach of a deep-sea isopod, Bathynomus jamesi. Enzyme kinetic studies of MyNal showed that it exhibited a very low Km for both cleavage and synthesis activities compared to previously described NALs. Though it favors the cleavage process, MyNal out-competes the known NALs with respect to the efficiency of Neu5Ac synthesis and exhibits the highest k(cat)/K(m)values. High expression levels of recombinant MyNal could be achieved (9.56 mol L-1 culture) with a stable activity in a wide pH (5.0-9.0) and temperature (40-60 degrees C) range. All these features indicated that the deep-sea NAL has potential in the industrial production of Neu5Ac. Furthermore, we found that the amino acid 189 of MyNal (equivalent to Phe190 in Escherichia coli NAL), located in the sugar-binding domain, GX189DE, was also involved in conferring its enzymatic features. Therefore, the results of this study improved our understanding of the NALs from different environments and provided a model for protein engineering of NAL for biosynthesis of Neu5Ac.
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WOS关键词	SIALIC-ACID METABOLISM ; D-NEURAMINIC ACID ; ESCHERICHIA-COLI ; CLOSTRIDIUM-PERFRINGENS ; HAEMOPHILUS-INFLUENZAE ; ENZYMATIC-SYNTHESIS ; EXPRESSION ; GENE ; PURIFICATION ; DIVERSITY
语种	英语
WOS记录号	WOS:000428510200006
内容类型	期刊论文
源URL	[http://ir.idsse.ac.cn/handle/183446/5966]
专题	深海科学研究部_深海生物学研究室_深海生物蛋白质组学及分子生物学研究组
通讯作者	He, Li-sheng; He, Li-sheng(Chinese Acad Sci, Inst Deep Sea Sci & Engn, Dept Life Sci, Sanya 572000, Peoples R China)
作者单位	1.Chinese Acad Sci, Inst Phys, Lab Soft Matter Phys, Beijing 100864, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Natl Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China 3.Univ Chinese Acad Sci, Coll Earth Sci, Beijing 100864, Peoples R China 4.Chinese Acad Sci, Inst Deep Sea Sci & Engn, Dept Life Sci, Sanya 572000, Peoples R China
推荐引用方式 GB/T 7714	Li, Yun-liang,Han, Zhuang,Chen, Xi,et al. Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma[J]. MARINE DRUGS,2018,16(3).
APA	Li, Yun-liang.,Han, Zhuang.,Chen, Xi.,Chen, Qi-jia.,Wang, Yong.,...&Wang, Shao-lu.(2018).Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma.MARINE DRUGS,16(3).
MLA	Li, Yun-liang,et al."Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma".MARINE DRUGS 16.3(2018).