极地研究

极地研究 ›› 2011, Vol. 22 ›› Issue (2-English): 124-130.DOI: 10.3724/SP.J.1085.2011.00124

• 研究论文 • 上一篇    

Cloning and heterologous expression of pro-2127, a gene encoding cold-active protease from Pseudoalteromonas sp. QI-1

XU GuoYing1,2, CUI ShuoShuo1,2 & LIN XueZheng1,2*   

  1. 1 Key Lab of Marine Bioactive Substances, SOA, Qingdao 266061, China;
    2 First Institute of Oceanography, SOA, Qingdao 266061, China
  • 收稿日期:2010-12-17 修回日期:2011-04-06 出版日期:1961-06-30 发布日期:1961-06-30
  • 通讯作者: LIN XueZheng
  • 基金资助:

    863计划重点项目“极地特殊海洋微生物资源利用的关键技术研究”;基本科研业务费专项资金项目（团队）“极地海冰微生物抗逆功能基因的研究与开发”

Cloning and heterologous expression of pro-2127, a gene encoding cold-active protease from Pseudoalteromonas sp. QI-1

XU GuoYing1,2, CUI ShuoShuo1,2 & LIN XueZheng1,2*   

  1. 1 Key Lab of Marine Bioactive Substances, SOA, Qingdao 266061, China;
    2 First Institute of Oceanography, SOA, Qingdao 266061, China
  • Received:2010-12-17 Revised:2011-04-06 Online:1961-06-30 Published:1961-06-30
  • Contact: LIN XueZheng

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摘要：

The psychrotropic bacterium, Pseudoalteromonas sp. QI-1, which produces extracellular cold-active protease, was isolated from Antarctic seawater. The genomic DNA of this bacterium was used to construct a plasmid genomic library with the goal of screening cold-active protease genes. Gene pro-2127 with an open reading frame of 2127 bp encoding protease PRO-2127 was cloned and sequenced. Alignment of amino acid sequences suggested that the precursor of PRO-2127 was a member of subfamily S8A, and that it might contain four domains: a signal peptide, an N-terminal prosequence, a catalytic domain and a C-terminal extension. Amino acids Asp185, His244 and Ser425 might form a catalytic triad. PRO-2127 showed some structural features common to psychrophilic enzymes, such as a decrease in Arg residues and the Arg/(Arg+Lys) ratio. Heterologous expression of pro-2127 in Escherichia coli BL21 (DE3) by pColdIII was also successfully observed in this study.

关键词: Antarctic, Pseudoalteromonas, cold-active protease, gene cloning and expression

Abstract:

The psychrotropic bacterium, Pseudoalteromonas sp. QI-1, which produces extracellular cold-active protease, was isolated from Antarctic seawater. The genomic DNA of this bacterium was used to construct a plasmid genomic library with the goal of screening cold-active protease genes. Gene pro-2127 with an open reading frame of 2127 bp encoding protease PRO-2127 was cloned and sequenced. Alignment of amino acid sequences suggested that the precursor of PRO-2127 was a member of subfamily S8A, and that it might contain four domains: a signal peptide, an N-terminal prosequence, a catalytic domain and a C-terminal extension. Amino acids Asp185, His244 and Ser425 might form a catalytic triad. PRO-2127 showed some structural features common to psychrophilic enzymes, such as a decrease in Arg residues and the Arg/(Arg+Lys) ratio. Heterologous expression of pro-2127 in Escherichia coli BL21 (DE3) by pColdIII was also successfully observed in this study.

Key words: Antarctic, Pseudoalteromonas, cold-active protease, gene cloning and expression