农业生物技术学报2001，Vol.9Issue(1)：81-85,5.
木霉 -1,3-1,4-葡聚糖酶性质及其cDNA片段克隆
Study on the Properties of -1,3-1,4-glucanase;Cloning and Sequence Analysis of cDNA Fragment from Trichoderma reesei
孙建义 1李卫芬 1许梓荣 1廖玉华1
作者信息
- 1. 浙江大学华家池校区动物科学学院饲料科学研究所,
- 折叠
摘要
Abstract
-1,3-1,4-glucanase was purified from a solid-state culture of Trichoderma reesei GXC on wheat bran in three steps which comprisedammonium sulfate precipitation,Sephadex G-100 chromatography,and DEAE-Sephadex A-50 chromatography.The molecular mass was determined to be 35.21 kilodaltons by 12.5%sodium dodecyl sulfate polyacrylamide gel electrophoresis.The optimal temperature and pH for purified -1,3-1,4-glucanase reaction were 50℃ and 6.0,respectively.The Km of the enzyme on -glucan was 10.86 mg/mL,and the Vmax on -glucan was 14 286 molof glucose equivalents per mg of the pure enzyme per min.A partial -1,3-1,4-glucanase cDNA Fragment from Trichoderma reesei GXC was amplified and cloned by Reverse Transcription-PCR strategy. Sequence analysis showed that the cDNA Fragment was 280 bp,it had 90%,79%,91% identity with the sequences of -1,3-1,4-glucanase from Bacillus amyloliquefaciens,Orpinomyces Strain PC-2 and Bacillus subtilis,respectively.关键词
里氏木霉GXC/-1,3-1,4-葡聚糖酶/酶学性质/克隆/测序分类
农业科技引用本文复制引用
孙建义,李卫芬,许梓荣,廖玉华..木霉 -1,3-1,4-葡聚糖酶性质及其cDNA片段克隆[J].农业生物技术学报,2001,9(1):81-85,5.
