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来源于瘤胃厌氧真菌Neocallimastixfrontalis木聚糖酶在毕赤酵母中的表达

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.05.029, PP. 186-193

Keywords: Neocallimastixfrontalis,木聚糖酶,毕赤酵母,密码子优化,酶学性质

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Abstract:

厌氧真菌Neocallimastixfrontalis是瘤胃中降解木聚糖和纤维素的主要微生物之一,其木聚糖酶具有潜在的应用价值。对来源于Neocallimastixfrontalis木聚糖酶基因Xyn11B进行密码子优化;通过全基因合成优化后的木聚糖酶基因Xyn11Bm,构建该基因的酵母表达载体pPIC9K-Xyn11Bm,并在毕赤酵母GS115中诱导表达。摇瓶水平时,重组Xyn11Bm酶活性最高为4874.8U/mL。在10L发酵罐中诱导96h后,重组Xyn11Bm的酶活性为5139.7U/mL,菌体湿重和干重达到216.7g/L和117.3g/L。酶学性质分析表明,重组Xyn11Bm的最适反应温度为50℃,最适反应pH为5.0。在pH5.0-8.0时该酶具有较好的稳定性,但温度稳定性较差。底物特异性分析表明,重组Xyn11Bm可水解燕麦木聚糖、桦木木聚糖和可溶性木聚糖4-O-Me-D-glucurono-D-xylan,但不降解地衣多糖和大麦β-葡聚糖。结果表明重组Xyn11Bm具有潜在的应用价值。

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