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微生物蛋白质谷氨酰胺酶的初步分离纯化

, PP. 161-165

Keywords: 微生物蛋白质谷氨酰胺酶,超滤,乙醇沉淀,离子交换层析,SDS-PAGE

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Abstract:

蛋白质谷氨酰胺酶可以特异性地水解蛋白质和多肽的谷氨酰胺残基,研究了产吲哚金黄杆菌产生的微生物蛋白质谷氨酰胺酶的代谢曲线和初步的分离纯化。发酵过程中pH升高,氨浓度增加,到14h时酶活达到最高为0.359U/mL。发酵液离心去上清液经3kD滤膜超滤4倍时,纯化倍数和得率都最高。超滤液加入4倍体积无水乙醇沉淀蛋白质,酶的得率最高为72.7%。沉淀用缓冲液复溶后,经过SP-SepharoseFastFlow离子交换层析分离,得到单一峰。经过多步纯化后酶得率为31.72%,纯化倍数为124倍,经SDS-PAGE电泳鉴定为单一条带,分子量约为20kD。

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