Keywords

Lipase, Jatropha Curcas, Degenerate Primers, Protein Structure

Abstract

Lipase (Lipase, EC3.1.1.3) is an industrial enzyme preparation commonly used in the leather, feed and biodiesel industries and has wide application value. There are currently few studies on plant-derived lipases. In this study, Jatropha curcas (Jatrophacurcas), an oil plant with promising application in biodiesel, was used as the research object, and the lipase gene (JcLIP) of this species was cloned. Through multiple sequence alignment and the genetic relationship of species, degenerate primers with higher specificity were designed. By using RT-PCR and RACE technology, the full-length sequence of the leprosy resinase gene was finally obtained and successfully expressed in E. coli Expressed in Escherichia coli, the enzyme activity measurement results show that leprosy resinase is expressed in inclusion bodies in E. coli, but can produce active proteins with an enzyme activity of about 0.8U.mL-1. Structural prediction and comparison show that JcLIP protein has the structural core and catalytic active center of lipase, but has more insertions and random coils in the non-core region than Mucor lipase, which may determine the difference in enzyme activity between the two. important reason.

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Original research done by Wang Xiaoxing, Peng Feng, Niu Dongyun, Li Gang, Chen Fang

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