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OALib Journal期刊
ISSN: 2333-9721
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嗜热子囊菌光孢变种内切葡聚糖酶i基因在毕赤酵母中的表达及部分酶学性质
DOI: 10.3724/SP.J.1145.2009.00419, PP. 419-422
Keywords: 纤维素酶,内切葡聚糖酶,嗜热子囊菌光孢变种,嗜热真菌,毕赤酵母
Abstract:
摘要嗜热子囊菌是一种嗜热真菌，可以产生具有很高工业价值的内切葡聚糖酶.本研究成功表达了嗜热子囊菌内切葡聚糖酶ⅰ基因，并获得热稳定的重组内切葡聚糖酶.提取嗜热子囊菌光孢变种(thermoascusaurantiacusvar.levisporus)总rna，通过rt-pcr方法克隆出内切-β-葡聚糖酶eg1基因的成熟肽编码序列.采用基因重组的方法构建该基因的巴斯德毕赤酵母pichiapastoris分泌型表达载体ppic9k-eg1，经线性化后采用电穿孔法将其导入毕赤酵母gs115中，大量筛选后获得高效表达内切葡聚糖酶ⅰ的毕赤酵母工程菌株gpn24.该菌株采用甲醇诱导120h后，内切葡聚糖酶ⅰ的活力可达570.7u/ml，最适温度为55℃，在90℃的条件下保温30min后仍具有60％的酶活力；最适ph为5.0，在ph3.0~5.0的条件下酶活力保持稳定.图6表2参16
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