Takada G, Kawaguchi T, Sumitani J, Arai M
Department of Applied Biological Chemistry, College of Agriculture, Osaka Prefecture University, Sakai, Japan.
Biosci Biotechnol Biochem. 1998 Aug;62(8):1615-8. doi: 10.1271/bbb.62.1615.
A cellobiohydrolase I (cbhI) and a beta-glucosidase 1 (bgl1) gene of Aspergillus aculeatus were expressed in Saccharomyces cerevisiae. The transformed cells secreted the enzymes efficiently in an active form. The recombinant CBHI gave two bands of different molecular mass (110 and 90 kDa) and the recombinant BGL1 gave one band (180 kDa) by SDS-PAGE. The recombinant CBHI and BGL1 had the same enzymatical properties as the native enzyme except for the specific activity toward cellulosic substrates. By the combination of three different types of cellulases, FI-CMCase, CBHI, and BGL1, we could hydrolyze Avicel up to 59% under our experimental conditions.
棘孢曲霉的纤维二糖水解酶I(cbhI)和β-葡萄糖苷酶1(bgl1)基因在酿酒酵母中表达。转化后的细胞能高效分泌活性形式的酶。通过SDS-PAGE分析,重组CBHI出现两条不同分子量的条带(110 kDa和90 kDa),重组BGL1出现一条条带(180 kDa)。重组CBHI和BGL1除了对纤维素底物的比活性外,具有与天然酶相同的酶学性质。在我们的实验条件下,通过将三种不同类型的纤维素酶,即FI-CMCase、CBHI和BGL1组合使用,我们可以将微晶纤维素水解高达59%。