Cloning And Expression Of Lipase Gene From Rhizopus Delemar

Lipase from Rhizopus delemar has 1,3 positional specificity, it can hydrolyze triacylglycerol produced fatty acid and monoglyceride(MG). MG is a important non-ionic surfactants, widely used as emulsifier in the food stuff and other industries. However the high operation cost, poor stability and low activity restrict its application. Today people focus on constructing gene strain of high enzyme activity.The gene which encoding lipase from Rhizopus delemar AS3.230 was amplified by PCR and cloned into vector pUCm-T. The recombinant plasmid named pUCm-T-LipRD, and transferred into E.coli DH5a. The pUCm-T-LipRD sequenced with dideoxynucleotide termination method.After the DNA sequence was determined, the gene was coloned into the yeast integrative plasmid pPIC9K, which was then transformed into His4 mutant yeast GS115. Recombinant Pichia strains were obtained by G418 plates and tributyrin flat screening, and obtained seven high expression recombinant. The engineered strain was expressed by the induction of methanol. The expression products of RDL gene was...