Identification, Gene Expression and Enzyme Characterization of Serratia sp.Producing Cold Adapted Lipase | |
Liu, Yuanli; Chen, Jixiang; Li, Yanlin; Zhou, Min; Cheng, Lin | |
刊名 | Journal of Chinese Institute of Food Science and Technology |
2018-06-30 | |
卷号 | 18期号:6页码:121-129 |
关键词 | Affinity chromatography Bacteria Biodiesel Cell proliferation Cloning Genes Polymerase chain reaction Recombinant proteins Synthetic fuels Catalytic synthesis Cold-adapted lipase DNA sequence analysis Enzymatic activities Enzyme characterization Metal-affinity chromatography Morphological characteristic Serratia |
ISSN号 | 10097848 |
DOI | 10.16429/j.1009-7848.2018.06.016 |
英文摘要 | A bacterium LHY-1 producing cold-adapted lipase was isolated from the contaminated oil samples in Qinghai Province by the method of Rhodamine B plate. The strain was preliminarily identified as Serratia sp. by morphological characteristics, physiological and biochemical, and 16 S r DNA sequence analysis. The optimum carbon source, nitrogen source, temperature, p H for the cell growth and lipase production were beef extract, yeast powder, 30 and p H7.0 respectively. The bacterium could grow in the medium with salinity of 9%. The lipase production was 1 326 U/m L under optimum culture conditions. A lipase gene was cloned from LHY-1 by PCR amplification with specific primers of the lipase of Serratia. An expression vector p ET-28 a (+) /lipase was constructed. The expressed lipase was purified by NiNTA metal affinity chromatography. The recombinant protein was 36 ku on SDS-PAGE. The optimal temperature for the recombinant lipase activity was at 30 . The lipase was stable at temperature range of 0-30 and p H range of 7-11. It belonged to cold active lipase. Addition of Mg2+and Ca2+increased its activity, but Co2+, Zn2+, PMSF, SDS, EGTA and EDTA inhibited the enzymatic activity. The lipase can be used in catalytic synthesis of biodiesel. Keyword: © 2018, Editorial Office of Journal of CIFST. All right reserved. |
语种 | 中文 |
出版者 | Chinese Institute of Food Science and Technology |
内容类型 | 期刊论文 |
源URL | [http://ir.lut.edu.cn/handle/2XXMBERH/114072] |
专题 | 石油化工学院 |
作者单位 | School of Petrochemical Engineering, Lanzhou University of Technology, Lanzhou; 730050, China |
推荐引用方式 GB/T 7714 | Liu, Yuanli,Chen, Jixiang,Li, Yanlin,et al. Identification, Gene Expression and Enzyme Characterization of Serratia sp.Producing Cold Adapted Lipase[J]. Journal of Chinese Institute of Food Science and Technology,2018,18(6):121-129. |
APA | Liu, Yuanli,Chen, Jixiang,Li, Yanlin,Zhou, Min,&Cheng, Lin.(2018).Identification, Gene Expression and Enzyme Characterization of Serratia sp.Producing Cold Adapted Lipase.Journal of Chinese Institute of Food Science and Technology,18(6),121-129. |
MLA | Liu, Yuanli,et al."Identification, Gene Expression and Enzyme Characterization of Serratia sp.Producing Cold Adapted Lipase".Journal of Chinese Institute of Food Science and Technology 18.6(2018):121-129. |
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