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菌物学报  2015 

嗜热毛壳菌热稳定脂肪酶基因(lm)的克隆及其在毕赤酵母中的表达

DOI: DOI:10.13346/j.mycosystema.140046, PP. 434-442

Keywords: 嗜热真菌,分子量,酶活,最适温度,最适pH,热稳定性

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Abstract:

研究从嗜热毛壳菌Chaetomiumthermophilum中克隆了一个新的脂肪酶基因(lm)。其中DNA序列包含一个由870个碱基构成的开放阅读框,编码289个氨基酸,含有4个内含子,没有信号肽序列。序列提交GenBank,登录号为GU338248。将该基因在毕赤酵母中表达。在甲醇的诱导下,重组蛋白得到了高效表达,第6天的表达量最高,蛋白达到0.428mg/mL,酶活力为19.77U/mg。SDS-PAGE检测该蛋白的分子量为35kDa。该脂肪酶的最适反应温度为60℃,具有热稳定性,在40–80℃热稳定,80℃处理60min仍有65%的相对酶活。该酶最适反应pH值为10.0,在pH9.0–12.0酶活相对稳定。该酶具有较好的热稳定性和耐碱性,具有良好的工业应用价值。

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