变形杆菌属脂肪酶基因的克隆表达及酶学性质研究
首发时间:2012-11-12
摘要:筛选获得一株产脂肪酶的菌株,经16S rDNA 鉴定属于变形杆菌属。根据已报道的来源于变形杆菌的脂肪酶基因设计引物,克隆其全长基因,命名为lipK19,大小为864 bp,编码287个氨基酸。构建重组表达质粒pET32a-lipK19,在大肠杆菌中表达酶活可达1500 U/L。该脂肪酶的最适温度为40 ℃,最适pH为9,在pH 8~10之间稳定性较好,Ca2+对酶活有激活作用,表面活性剂等对酶活抑制作用明显,对有机溶剂有一定的耐受性。
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Cloning, Expression and Characterization of the Lipase LipK19 from a screened Proteus sp
Abstract:A lipase production strain identified as Proteus sp by 16S rDNA was screened and named as K19. According to the reported lipase gene from Proteus vulgaris K80, a pair of primers was designed to clone the full length lipase gene of K19, which was named as lipK19 with the size of 864 bp, and encoding 287 amino acids. Recombinant expression plasmid pET32a-lipK19 was constructed, which was transferred in E. coli and induced with IPTG, the expressed LipK19 activity attained to 1500 U/L. The optimum temperature of LipK19 was 40 ℃, and the optimum pH was 9 with high stability between pH 8~10. The Ca2+could activate the activity of LipK19, but the surfactants inhibited the enzyme activity significantly. This lipase showed a certain degree of tolerance to organic solvent.
Keywords: Proteus sp Lipase Cloning Expression Characterization
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