Cloning of a gene encoding a thermo-stable endo-beta-1,4-glucanase from Thermoascus aurantiacus and its expression in yeast.
A gene encoding a thermo-stable endo-beta-1,4-glucanase was isolated from the thermophilic fungus, Thermoascus aurantiacus IFO9748, and designated as eg1. Induction of this gene expression at 50 degrees C was stronger than at 30 degrees 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 70 degrees C and the product was stable from pH 3-10. The enzyme was optimally active at 70 degrees C over 20 min and the optimal pH was 6.
Hong
Jiong
J
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
Tamaki
Hisanori
H
Yamamoto
Kenji
K
Kumagai
Hidehiko
H
D002478:Cells, Cultured; D002480:Cellulase / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D003001:Cloning, Molecular / Q000379:methods; D004789:Enzyme Activation; D004795:Enzyme Stability; D032641:Eurotiales / Q000201:enzymology* / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D013696:Temperature
D011994:Recombinant Proteins; D002480:Cellulase