Department of Biotechnology, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.
J Agric Food Chem. 2012 Mar 7;60(9):2354-61. doi: 10.1021/jf2049799. Epub 2012 Feb 22.
Production, purification, and characterization of a novel β-1,3-1,4-glucanase (lichenase) from thermophilic Rhizomucor miehei CAU432 were investigated. High-level extracellular β-1,3-1,4-glucanase production of 6230 U/mL was obtained when oat flour (3%, w/v) was used as a carbon source at 50 °C. The crude enzyme was purified to homogeneity with a specific activity of 28818 U/mg. The molecular weight of purified enzyme was estimated to be 35.4 kDa and 33.7 kDa by SDS-PAGE and gel filtration, respectively. The optimal pH and temperature of the enzyme were pH 5.5 and 60 °C, respectively. The K(m) values of purified β-1,3-1,4-glucanase for barley β-glucan and lichenan were 2.0 mM and 1.4 mM, respectively. Furthermore, the gene (RmLic16A) encoding the β-1,3-1,4-glucanase was cloned and its deduced amino acid sequence showed the highest identity (50%) to characterized β-1,3-1,4-glucanase from Paecilomyces thermophila. The high-level production and biochemical properties of the enzyme enable its potential industrial applications.
研究了嗜热根毛霉 CAU432 中一种新型β-1,3-1,4-葡聚糖酶（lichenase）的生产、纯化和特性。当以燕麦粉（3%，w/v）作为碳源在 50°C 下使用时，获得了 6230 U/mL 的高水平胞外β-1,3-1,4-葡聚糖酶产量。粗酶经纯化后比活为 28818 U/mg，达到均一性。SDS-PAGE 和凝胶过滤分别估计纯化酶的分子量为 35.4 kDa 和 33.7 kDa。酶的最适 pH 和温度分别为 pH 5.5 和 60°C。纯化的β-1,3-1,4-葡聚糖酶对大麦β-葡聚糖和lichenan 的 K(m)值分别为 2.0 mM 和 1.4 mM。此外，还克隆了编码β-1,3-1,4-葡聚糖酶的基因（RmLic16A），其推导的氨基酸序列与已鉴定的嗜热拟青霉β-1,3-1,4-葡聚糖酶具有最高的同源性（50%）。该酶的高水平生产和生化特性使其具有潜在的工业应用前景。
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