期刊
SYSTEMATIC AND APPLIED MICROBIOLOGY
卷 36, 期 3, 页码 197-204出版社
ELSEVIER GMBH, URBAN & FISCHER VERLAG
DOI: 10.1016/j.syapm.2012.12.008
关键词
Lipase; Metagenomic library; Gene cloning; Expression; Pichia pastoris
资金
- National Natural Science Foundation of China [30800039]
- Institute of Pathogen Biology, Chinese Academy of Medical Sciences [2007IPB007]
Lipases are widely used for a variety of biotechnological applications. Screening these industrial enzymes directly from environmental microorganisms is a more efficient and practical approach than conventional cultivation-dependent methods. Combined with activity-based functional screening, six clones with lipase activity were detected and a gene (termed lipZ01) isolated from a target clone with the highest lipase activity was cloned from an oil-contaminated soil-derived metagenomic library and then sequenced. Gene lipZ01 was expressed in Pichia pastoris GS115 and the molecular weight of the recombinant lipase LipZ01 was estimated by electrophoresis analysis to be approximately 50 kDa. The maximum activity of the purified lipase was 42 U/mL, and the optimum reaction temperature and pH value were 45 degrees C and 8.0, respectively. The enzyme was highly stable in the temperature range 35-60 degrees C and under alkaline conditions (pH 7-10). The presence of Ca2+ and Mn2+ ions could significantly enhance the activity of the lipase. The purified lipase preferentially hydrolysed triacylglycerols with acyl chain lengths >= 8 carbon atoms, and the conversion degree of biodiesel production was nearly 92% in a transesterification reaction using olive oil and methanol. Some attractive properties suggested that the recombinant lipase may be valuable in industrial applications. (C) 2013 Elsevier GmbH. All rights reserved.
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