从绿色木霉中克隆两个纤维二糖水解酶基因，并在酵母酿酒酵母中异源表达。
Cloning of two cellobiohydrolase genes from Trichoderma viride and heterogeneous expression in yeast Saccharomyces cerevisiae.
机构信息
Department of Life Science and Engineering, Harbin Institute of Technology, P.O. Box 437, No. 92, West Da-Zhi Street, Harbin, Heilongjiang, 150001, China.
出版信息
Mol Biol Rep. 2010 Apr;37(4):2135-40. doi: 10.1007/s11033-009-9683-3. Epub 2009 Aug 11.
Cellobiohydrolase genes cbhI and cbhII were isolated from Trichoderma viride AS3.3711 and T. viride CICC 13038, respectively, using RT-PCR technique. The cbhI gene from T. viride AS3.3711 contains 1,542 nucleotides and encodes a 514-amino acid protein with a molecular weight of approximately 53.96 kDa. The cbhII gene from T. viride CICC 13038 was 1,413 bp in length encoding 471 amino acid residues with a molecular weight of approximately 49.55 kDa. The CBHI protein showed high homology with enzymes belonging to glycoside hydrolase family 7 and CBHII is a member of Glycoside hydrolase family 6. CBHI and CBHII play a role in the conversion of cellulose to glucose by cutting the disaccharide cellobiose from the non-reducing end of the cellulose polymer chain. The two cellobiohydrolase (CBHI, CBHII) genes were successfully expressed in Saccharomyces cerevisiae H158. Maximal activities of transformants Sc-cbhI and Sc-cbhII were 0.03 and 0.089 units ml(-1) under galactose induction, respectively. The optimal temperatures of the recombinant enzymes (CBHI, CBHII) were 60 and 70 degrees C, respectively. The optimal pHs of recombinant enzymes CBHI and CBHII were at pH 5.8 and 5.0, respectively.
纤维二糖水解酶基因 cbhI 和 cbhII 分别从绿色木霉 AS3.3711 和绿色木霉 CICC 13038 中通过 RT-PCR 技术分离得到。来自绿色木霉 AS3.3711 的 cbhI 基因包含 1542 个核苷酸，编码一个 514 个氨基酸的蛋白质，分子量约为 53.96 kDa。来自绿色木霉 CICC 13038 的 cbhII 基因长度为 1413bp，编码 471 个氨基酸残基，分子量约为 49.55 kDa。CBHI 蛋白与属于糖苷水解酶家族 7 的酶具有高度同源性，而 CBHII 是糖苷水解酶家族 6 的成员。CBHI 和 CBHII 通过从纤维素聚合物链的非还原端切割二糖纤维二糖来将纤维素转化为葡萄糖。两个纤维二糖水解酶（CBHI、CBHII）基因在酿酒酵母 H158 中成功表达。在半乳糖诱导下，转化子 Sc-cbhI 和 Sc-cbhII 的最大酶活分别为 0.03 和 0.089 单位 ml(-1)。重组酶（CBHI、CBHII）的最适温度分别为 60 和 70°C。重组酶 CBHI 和 CBHII 的最适 pH 分别为 pH5.8 和 pH5.0。
Zotero 插件