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

文献类型：期刊论文

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作者Wang, Shao-lu1,2; Li, Yun-liang3; Han, Zhuang1; Chen, Xi4; Chen, Qi-jia4; Wang, Yong1; He, Li-sheng1; He, Li-sheng(Chinese Acad Sci, Inst Deep Sea Sci & Engn, Dept Life Sci, Sanya 572000, Peoples R China)
刊名MARINE DRUGS ; MARINE DRUGS
出版日期2018-03-01 ; 2018-03-01
卷号16期号:3
关键词N-acetylneuraminate Lyase N-acetylneuraminate Lyase Symbiotic Microbe N-acetylneuraminic Acid Mycoplasma Symbiotic Microbe N-acetylneuraminic Acid Mycoplasma
DOI10.3390/md16030080 ; 10.3390/md16030080
产权排序第1完成单位 ; 第1完成单位 ; 第2完成单位 ; 第3完成单位 ; 第4完成单位 ; 第2完成单位 ; 第3完成单位 ; 第4完成单位
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)
文献子类Article ; 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.; 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.
URL标识查看原文 ; 查看原文
WOS关键词SIALIC-ACID METABOLISM ; SIALIC-ACID METABOLISM ; D-NEURAMINIC ACID ; ESCHERICHIA-COLI ; CLOSTRIDIUM-PERFRINGENS ; HAEMOPHILUS-INFLUENZAE ; ENZYMATIC-SYNTHESIS ; EXPRESSION ; GENE ; PURIFICATION ; DIVERSITY ; D-NEURAMINIC ACID ; ESCHERICHIA-COLI ; CLOSTRIDIUM-PERFRINGENS ; HAEMOPHILUS-INFLUENZAE ; ENZYMATIC-SYNTHESIS ; EXPRESSION ; GENE ; PURIFICATION ; DIVERSITY
语种英语 ; 英语
WOS记录号WOS:000428510200006 ; WOS:000428510200006
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 Deep Sea Sci & Engn, Dept Life Sci, Sanya 572000, Peoples R China
2.Univ Chinese Acad Sci, Coll Earth Sci, Beijing 100864, Peoples R China
3.Chinese Acad Sci, Inst Phys, Lab Soft Matter Phys, Beijing 100864, Peoples R China
4.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Natl Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China
推荐引用方式
GB/T 7714		 Wang, Shao-lu,Li, Yun-liang,Han, Zhuang,et al. Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma, Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma[J]. MARINE DRUGS, MARINE DRUGS,2018, 2018,16, 16(3).
APA Wang, Shao-lu.,Li, Yun-liang.,Han, Zhuang.,Chen, Xi.,Chen, Qi-jia.,...&He, Li-sheng.(2018).Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma.MARINE DRUGS,16(3).
MLA Wang, Shao-lu,et al."Molecular Characterization of a Novel N-Acetylneuraminate Lyase from a Deep-Sea Symbiotic Mycoplasma".MARINE DRUGS 16.3(2018).

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来源：深海科学与工程研究所

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