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毕赤酵母表达重组葡萄糖氧化酶的发酵条件

, PP. 136-141

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Abstract:

对甲醇诱导重组PichiapastorisGS115mut+高效表达黑曲霉葡萄糖氧化酶基因的条件进行了研究,拟建立一条高效低成本的生产重组葡萄糖氧化酶的工艺路线。通过摇瓶试验对发酵培养基、诱导时机、pH、诱导温度、甲醇诱导浓度等进行了优化,随后进行了50L发酵罐扩大化培养。结果表明,最适发酵培养基为YEPD培养基,甲醇诱导产酶的最佳时机为菌体对数生长末期,最佳诱导浓度为1%,最适诱导pH范围为6-6.5,最适诱导温度25℃。按优化的条件用50L发酵罐分批培养酵母14h,甲醇诱导培养48h后生物量和酶活性达到最高。菌体终密度达到OD600为44,每升发酵液中产酶105kU,发酵液上清蛋白含量为1000mg/L。

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