Recombinant pichia pastoris for heterogenous efficient expression of lipase and application of recombinant pichia pastoris

Abstract

The invention discloses recombinant pichia pastoris for heterogenous efficient expression of lipase and application of the recombinant pichia pastoris. The recombinant pichia pastoris is obtained by converting plasmid HAC1-pPIC3.5K of an overexpression HAC1 gene in recombinant pichia pastoris X-33 which contains a pro-rml gene of a copy number 4 and is capable of expressing Pro-RML. When the strain is subjected to 96 hours of flask shaking fermentation, the extracellular enzyme activity is up to 1078U / mL at most, and the enzyme activity secretion efficiency is up to 47U / OD600. 120 hours of fermentation is needed when pichia pastoris which is disclosed by patent CN103361327A and has a rhizomucor mieheilipase copy number of 2 meets the highest extracellular enzyme activity and enzyme secretion efficiency, the extracellular enzyme activity is 1038U / mL at most, and the enzyme activity secretion efficiency is only 25U / OD600. By adopting the recombinant pichia pastoris, expression of rhizomucor mieheilipase is effectively promoted, on the premise that the highest extracellular enzyme activity is not influenced, the secretion efficiency of the rhizomucor mieheilipase is increased by 1.9 times, and the fermentation time is shortened by 24 hours.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and relates to a strain of recombinant Pichia pastoris expressing lipase with heterologous high-efficiency and application thereof. Background technique

[0002] Lipase (E C3.1.1.3) is a class of enzymes that can hydrolyze triacylglycerol. It can catalyze a variety of chemical reactions such as: hydrolysis, esterification, transesterification, aminolysis, etc., and has mild and specific reaction conditions. Strong, non-polluting and other advantages are widely used in food, medicine, cosmetics, petroleum and other industries, known as the third largest industrial enzymes.

[0003] Although lipase has a wide range of uses, the low yield of the starting strain limits its application. Therefore, heterologous protein expression is currently the main strategy for industrial protein production. However, there are many factors that affect the expression of heterologous proteins, such as host selection, gen...

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