Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
| Cloning and characterization of a leucine aminopeptidase from Pseudoalteromonas telluritireducens DSM 16098, a strain isolated from hydrothermal vents fluid | |
| Zhang, Huan1,4; Wang, Hao1,4; Liu, Rui2,4; Wang, Lingling3; Song, Linsheng3 | |
| 2018-08-01 | |
| 发表期刊 | DEEP-SEA RESEARCH PART I-OCEANOGRAPHIC RESEARCH PAPERS
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| ISSN | 0967-0637 |
| 卷号 | 138页码:114-121 |
| 通讯作者 | Wang, Hao([email protected]) ; Wang, Lingling([email protected]) |
| 摘要 | Microbes living around hydrothermal vents have developed lots of enzymes with specific characteristics that allow them to thrive in the environment. Extracellular enzyme leucine aminopeptidases (LAPs) are all metallopeptidases that catalyze the hydrolysis of leucine residues from N-terminal residues from peptides and proteins. Genomic analysis of Pseudoalteromonas telluritireducens DSM 16098, a strain of deep-sea bacteria isolated from hydrothermal vents fluid, revealed the presence of an LAP gene (designated as PtLAP). The open reading frame of PtLAP encodes a protein of 614 amino acids (GenBank accession no. KY429939), consisting of a signal peptide, a peptidase_M1 domain and a leuk-A4-hydro_C domain. The putative substrate binding site GGMEN, zinc-binding motif HEXXHX18E and the catalytic residues involved in aminopeptidase activity are all conserved, indicating PtLAP is a member of M1 leucine aminopeptidases. The recombinant PtLAP expressed in Escherichia coli Transetta (DE3) (designated as rPtLAP) exhibits a significant LAP activity to hydrolyze L-leucine 7-amido-4-methylcoumarin (MAC-L). The optimal activity of rPtLAP is observed at pH 7.2 and 28 degrees C with half-saturation coefficient (K-m) of 44.56 +/- 5.09 mu mol L-1. rPtLAP activity is relatively stable at 30-40 degrees C and exhibits a relatively wide pH range (6.0-12.5) with the maximal activity ranging at pH 7.2-8.0. It can be inhibited by EDTA, protease inhibitors and some metal ions including Cu2+, Zn2+ and Cd2+, but is relatively tolerant to Mn2+ which is enriched in hydrothermal vent fluids. Furthermore, rPtLAP can significantly promote the proliferation of tumor cells, including the cell lines HCT116, BEL-7402 and Hela, while displays no effect on that of normal cell 3T3. These results together suggest that the extracellular enzyme PtLAP executes the degradation of organic matters, contributes to the adaptive survival of microbe in deep-sea environment and may play important roles in marine biogeochemical cycles. |
| 关键词 | Leucine aminopeptidases Pseudoalteromonas telluritireducens Recombinant protein Enzyme activity Cell proliferation |
| DOI | 10.1016/j.dsr.2018.06.006 |
| 收录类别 | SCI |
| 语种 | 英语 |
| 资助项目 | Chinese Academy of Sciences[XDA11030202]###983; Chinese Ministry of Science and Technology[2014AA093501]###984; Chinese Academy of Sciences[XDA11030202]; Chinese Ministry of Science and Technology[2014AA093501] |
| WOS研究方向 | Oceanography |
| WOS类目 | Oceanography |
| WOS记录号 | WOS:000445985100010 |
| 出版者 | PERGAMON-ELSEVIER SCIENCE LTD |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qdio.ac.cn/handle/337002/160399 |
| 专题 | 实验海洋生物学重点实验室 |
| 通讯作者 | Wang, Hao; Wang, Lingling |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Ctr Deep Sea Res, 7 Nanhai Rd, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, 7 Nanhai Rd, Qingdao 266071, Peoples R China 3.Dalian Ocean Univ, Liaoning Key Lab Marine Anim Immunol, 52 Heishijiao St, Dalian 116023, Peoples R China 4.Chinese Acad Sci, Ctr Ocean Megasci, 7 Nanhai Rd, Qingdao 266071, Peoples R China |
| 第一作者单位 | 深海极端环境与生命过程研究中心; 中国科学院海洋大科学研究中心 |
| 通讯作者单位 | 深海极端环境与生命过程研究中心; 中国科学院海洋大科学研究中心 |
| 推荐引用方式 GB/T 7714 | Zhang, Huan,Wang, Hao,Liu, Rui,et al. Cloning and characterization of a leucine aminopeptidase from Pseudoalteromonas telluritireducens DSM 16098, a strain isolated from hydrothermal vents fluid[J]. DEEP-SEA RESEARCH PART I-OCEANOGRAPHIC RESEARCH PAPERS,2018,138:114-121. |
| APA | Zhang, Huan,Wang, Hao,Liu, Rui,Wang, Lingling,&Song, Linsheng.(2018).Cloning and characterization of a leucine aminopeptidase from Pseudoalteromonas telluritireducens DSM 16098, a strain isolated from hydrothermal vents fluid.DEEP-SEA RESEARCH PART I-OCEANOGRAPHIC RESEARCH PAPERS,138,114-121. |
| MLA | Zhang, Huan,et al."Cloning and characterization of a leucine aminopeptidase from Pseudoalteromonas telluritireducens DSM 16098, a strain isolated from hydrothermal vents fluid".DEEP-SEA RESEARCH PART I-OCEANOGRAPHIC RESEARCH PAPERS 138(2018):114-121. |
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| Cloning and characte（1328KB） | 期刊论文 | 出版稿 | 限制开放 | CC BY-NC-SA | 浏览 | |
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