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共表达内质网响应蛋白HAC1对α-半乳糖苷酶在毕氏酵母中表达的影响

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Keywords: HAC1,α-半乳糖苷酶,毕氏酵母,共表达,UPR反应

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Abstract:

为提高毕氏酵母(Pichiapastoris)中重组α-半乳糖苷酶的表达,将来源于P.pastorisGS115的hac1基因构建至pPIC9K表达载体中,转化到重组P.pastoris中与α-半乳糖苷酶共表达。研究结果表明,在AOX1启动子控制下,HAC1过表达提高了重组P.pastoris中α-半乳糖苷酶蛋白表达含量,使α-半乳糖苷酶活性提高了2.2倍。经PCR产物鉴定,结果表明重组P.pastoris基因组中插入hac1基因。经50L发酵罐甲醇诱导168h,Gal-HAC1-4#工程菌最终酶活达到6560U/mL,比Gal-21#工程菌提高了27%。甲醇诱导后Gal-HAC1-4#发酵液中粗蛋白浓度均高于Gal-21#工程菌。表明共表达HAC1蛋白可提高毕氏酵母产α-半乳糖苷酶蛋白的能力。

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