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Cloning of a gene encoding a thermo-stable endo-beta-1,4-glucanase from Thermoascus aurantiacus and its expression in yeast
Authors:Hong Jiong  Tamaki Hisanori  Yamamoto Kenji  Kumagai Hidehiko
Institution:(1) Division of Applied Life Sciences, Graduate School of Agriculture, Sciences, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan;(2) Division of Integrated Life Sciences, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan
Abstract:A gene encoding a thermo-stable endo-beta-1,4-glucanase was isolated from the thermophilic fungus, Thermoascus aurantiacusIFO9748, and designated as eg1. Induction of this gene expression at 50thinsp°C was stronger than at 30thinsp°C. The deduced amino acid sequence encoded by eg1 showed that it belongs to the glycoside hydrolase family 5. The cloned gene was expressed in Saccharomyces cerevisiae and the gene product was purified and characterized. No significant activity loss was detected over 2 h at 70thinsp°C and the product was stable from pH 3–10. The enzyme was optimally active at 70thinsp°C over 20 min and the optimal pH was 6.
Keywords:cloning  endo-beta-1" target="_blank">gif" alt="beta" align="MIDDLE" BORDER="0">-1  4-d-glucanase  expression  Thermoascus aurantiacus
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