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南极耐冷 Pseudomonas sp. 7323低温脂肪酶的性质及基因克隆
引用本文:王水琦,曾润颖,张金伟.南极耐冷 Pseudomonas sp. 7323低温脂肪酶的性质及基因克隆[J].中国生物化学与分子生物学报,2007,23(4):271-277.
作者姓名:王水琦  曾润颖  张金伟
作者单位:1. 漳州职业技术学院,福建,漳州,363000
2. 国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,福建,厦门,361005
摘    要: 南极微生物是筛选低温酶的良好来源,但尚未得到充分的研究与开发.低温脂肪酶具有广阔的应用前景,其基因结构特征也具有重要的研究意义. 本文对南极微生物开展了低温脂肪酶产生菌的筛选、基因克隆及特征分析.采用功能筛选的方法,从南极普里兹湾深海沉积物中获得一株产低温脂肪酶的菌株7323,其最适温度和最高生长温度分别为20℃和30℃,属于耐冷菌.16S rDNA序列分析表明,该菌属于假单胞菌属(Pseudomonas).通过设计引物扩增出的脂肪酶基因全长为1854 bp,该基因编码一个由617氨基酸、分子量预计为64466的蛋白质.氨基酸序列分析表明,该酶与Pseudomonas sp. UB48 的脂肪酶有89%的相似性,在催化区和C末端信号肽中存在高度保守的序列.纯化后的酶学性质研究表明,该脂肪酶的最适温度为35℃,最适pH值为9.0,为碱性低温酶.

关 键 词:南极  耐冷菌  假单胞菌  低温脂肪酶
收稿时间:2006-8-28
修稿时间:2006年8月28日

Characterization and Gene Cloning of Cold-active Lipase from Antarctic Psychrotrophic Pseudomonas sp. 7323
WANG Shui-Qi,ZENG Run-Ying,ZHANG Jin-Wei.Characterization and Gene Cloning of Cold-active Lipase from Antarctic Psychrotrophic Pseudomonas sp. 7323[J].Chinese Journal of Biochemistry and Molecular Biology,2007,23(4):271-277.
Authors:WANG Shui-Qi  ZENG Run-Ying  ZHANG Jin-Wei
Institution:Zhangzhou Institute of Technology, Zhangzhou 363000,Fujian,China; Key Laboratory of Marine Biogenetic Resources, State Oceanic Administration, Xiamen 361005,Fujian,China
Abstract:Microorganisms in Antarctic were good resource for screening the cold active enzymes, and were still inadequately studied and exploited. The cold active lipase (CAL) had good potential in the industry, and its gene characteristic was worth of study. Therefore, the screening of CAL- producing bacteria from Antarctic sample, the CAL gene clone, and the characterization of CAL were reported in this study. By using medium containing lipase substrates, a bacterium 7323 with high cold active lipase activity was isolated from the deep sea sediment of Prydz Bay, Antarctic. The optimal and highest temperatures for its growth were 20℃ and 30℃, respectively, indicating that it was a psychrotrophic bacterium. It was affiliated with the genus Pseudomonas by the analysis of 16S rDNA sequences. The Pseudomonas sp. 7323 lipase gene with 1 854 bp was cloned by PCR. It encoded a protein of 617 amino acids, exhibiting 89% of identity with the lipase amino acid sequences of Pseudomonas sp. UB48. Two conservative motifs were found in the catalytic site and C-terminal signal peptide domain. The optimal temperature and pH for the activity of purified lipase were 35℃ and pH 9.0, respectively, indicating that it belonged to alkaline cold active lipase.
Keywords:Antarctic  psychrotroph  Pseudomonas  cold active lipase
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