A novel protease-resistant alpha-galactosidase with high hydrolytic activity from Gibberella sp F75: gene cloning, expression, and enzymatic characterization
文献类型: 外文期刊
第一作者: Cao, Yanan
作者: Cao, Yanan;Wang, Yaru;Meng, Kun;Bai, Yingguo;Shi, Pengjun;Luo, Huiying;Yang, Peilong;Zhou, Zhigang;Yao, Bin;Zhang, Zhifang
作者机构:
关键词: alpha-Galactosidase;Gibberella;Protease-resistance;High hydrolytic activity
期刊名称:APPLIED MICROBIOLOGY AND BIOTECHNOLOGY ( 影响因子:4.813; 五年影响因子:4.697 )
ISSN: 0175-7598
年卷期: 2009 年 83 卷 5 期
页码:
收录情况: SCI
摘要: A novel alpha-galactosidase gene (aga-F75) from Gibberella sp. F75 was cloned and expressed in Escherichia coli. The gene codes for a protein of 744 amino acids with a 24-residue putative signal peptide and a calculated molecular mass of 82.94 kDa. The native structure of the recombinant Aga-F75 was estimated to be a trimer or tetramer. The deduced amino acid sequence showed highest identity (69%) with an alpha-galactosidase from Hypocrea jecorina (Trichoderma reesei), a member of the glycoside hydrolase family 36. Purified recombinant Aga-F75 was optimally active at 60A degrees C and pH 4.0 and was stable at pH 3.0-12.0. The enzyme exhibited broad substrate specificity and substantial resistance to neutral and alkaline proteases. The enzyme K (m) values using pNPG, melibiose, stachyose, and raffinose as substrates were 1.06, 1.75, 54.26, and 8.23 mM, respectively. Compared with the commercial alpha-galactosidase (Aga-A) from Aspergillus niger var. AETL and a protease-resistant alpha-galactosidase (Aga-F78) from Rhizopus sp. F78, Aga-F75 released 1.4- and 4.9-fold more galactose from soybean meal alone, respectively, and 292.5- and 8.6-fold more galactose from soybean meal in the presence of trypsin, respectively. The pH and thermal stability and hydrolytic activity of Aga-F75 make it potentially useful in the food and feed industries.
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