| 冰川土壤来源Brachybacterium sp.DB5低温脂肪酶基因的克隆与表达 |
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| 引用本文： | 刘万利,张国华,陈强,杨培龙,张宇宏,姚斌.冰川土壤来源Brachybacterium sp.DB5低温脂肪酶基因的克隆与表达[J].中国农业科技导报,2009,11(6):55-62. |
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| 作者姓名： | 刘万利 张国华 陈强 杨培龙 张宇宏 姚斌 |
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| 作者单位： | (1.兰州大学生命科学学院生物化学与分子生物学研究所, 兰州 730000,2.中国农业科学院饲料研究所农业部饲料生物技术重点实验室, 北京 100081,3.中国农业科学院生物技术研究所, 北京 100081) |
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| 基金项目： | 农业微生物菌种资源整理整合与共享项目，国家863计划项目，中国农业科学院生物技术研究所基本科研业务费专项 |
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| 摘 要： | 从新疆一号冰川土壤来源的54株低温细菌中筛出11株产脂肪酶菌株,其中一株短状杆菌Brachybacterium sp. DB5产脂肪酶活力较高。通过同源克隆和TAIL-PCR技术从Brachybacterium sp. DB5 基因组DNA中克隆得到一个脂肪酶基因LipDB5。LipDB5基因全长933 bp,编码310个氨基酸和一个终止密码子,理论蛋白分子质量为34.8 kDa,无信号肽序列。将LipDB5基因在大肠杆菌中进行重组表达,重组脂肪酶最适反应温度30℃,在5℃时能保持最高活力的34.7%,对热不稳定,60℃ 处理 30 min 剩余17.4 %酶活。研究结果表明LipDB5具有低温脂肪酶的性质,在低温生物催化领域具有潜在的应用前景。
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| 关 键 词： | 冰川土壤 短状杆菌 Brachybacterium 低温脂肪酶 基因表达 |
| 收稿时间： | 2009-04-13 |
| 修稿时间： | 2009-08-11 |
Gene Cloning and Expression of a Cold-adapted Lipase from Brachybacterium sp. DB5 Isolated from Glacier Soil |
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| LIU Wan-li,ZHANG Guo-hua,CHEN Qiang,YANG Pei-long,ZHANG Yu-hong,YAO Bin.Gene Cloning and Expression of a Cold-adapted Lipase from Brachybacterium sp. DB5 Isolated from Glacier Soil[J].Journal of Agricultural Science and Technology,2009,11(6):55-62. |
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| Authors: | LIU Wan-li ZHANG Guo-hua CHEN Qiang YANG Pei-long ZHANG Yu-hong YAO Bin |
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| Institution: | (1.Institute of Biochemistry and Molecular Biology, School of Life Science, Lanzhou University, Lanzhou 730000|2.Key Laboratory of Feed Biotechnology, Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081|3.Biotechnology Research Institute, CAAS, Beijing 100081, China) |
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| Abstract: | Eleven lipase-producing bacterial strains were screened from 54 cold-adapted strains, which were isolated from No.1 Glacier soil of Xinjiang. Among them, one strain identified as Brachybacterium sp. DB5 according to 16S rDNA sequence analysis, showed the highest lipolytic activity. The lipase gene LipDB5 with 933 bp was cloned from Brachybacterium sp. DB5 using TAIL-PCR technology. LipDB5 encoded 310 amino acid and one stop codon with a theoretical molecular weight of 34.8 kDa, but without signal peptide. The lipase gene was functionally expressed in E.coli BL21 (DE3). The recombinant LipDB5 was most active at 30 ℃, kept 34.7% of activity at 5 ℃. Moreover, the recombinant enzyme retained 17.4% of the activity after incubation at 60℃ for 30 min. These activity data indicated that LipDB5 had some cold-adapted lipase characteristics. As a new cold-adapted lipase, LipDB5 has the potential application value in the field of bio-catalysis. |
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| Keywords: | Brachybacterium |
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