从嗜热毁丝霉中克隆、纯化和表征半乳甘露聚糖降解酶。
Cloning, purification, and characterization of galactomannan-degrading enzymes from Myceliophthora thermophila.
机构信息
Chemical Faculty, Lomonosov Moscow State University, Moscow, 119991, Russia.
出版信息
Biochemistry (Mosc). 2012 Nov;77(11):1303-11. doi: 10.1134/S0006297912110090.
Genes of β-mannosidase 97 kDa, GH family 2 (bMann9), β-mannanase 48 kDa, GH family 5 (bMan2), and α-galactosidase 60 kDa, GH family 27 (aGal1) encoding galactomannan-degrading glycoside hydrolases of Myceliophthora thermophila C1 were successfully cloned, and the recombinant enzymes were purified to homogeneity and characterized. bMann9 displays only exo-mannosidase activity, the K(m) and k(cat) values are 0.4 mM and 15 sec(-1) for p-nitrophenyl-β-D-mannopyranoside, and the optimal pH and temperature are 5.3 and 40°C, respectively. bMann2 is active towards galactomannans (GM) of various structures. The K(m) and k(cat) values are 1.3 mg/ml and 67 sec(-1) for GM carob, and the optimal pH and temperature are 5.2 and 69°C, respectively. aGal1 is active towards p-nitrophenyl-α-D-galactopyranoside (PNPG) as well as GM of various structures. The K(m) and k(cat) values are 0.08 mM and 35 sec(-1) for PNPG, and the optimal pH and temperature are 5.0 and 60°C, respectively.
β-甘露聚糖酶 97 kDa、GH 家族 2（bMann9）、β-甘露聚糖酶 48 kDa、GH 家族 5（bMan2）和α-半乳糖苷酶 60 kDa、GH 家族 27（aGal1）的基因被成功克隆，这些基因编码嗜热毁丝霉 C1 降解半乳甘露聚糖的糖苷水解酶。重组酶被纯化至均一性，并进行了表征。bMann9 仅显示外切甘露糖苷酶活性，其对 p-硝基苯-β-D-甘露吡喃糖苷的 K(m)和 k(cat)值分别为 0.4 mM 和 15 sec(-1)，最适 pH 和温度分别为 5.3 和 40°C。bMan2 对各种结构的半乳甘露聚糖（GM）均具有活性。其对瓜尔豆胶的 K(m)和 k(cat)值分别为 1.3 mg/ml 和 67 sec(-1)，最适 pH 和温度分别为 5.2 和 69°C。aGal1 对 p-硝基苯-α-D-半乳糖吡喃糖苷（PNPG）以及各种结构的 GM 均具有活性。其对 PNPG 的 K(m)和 k(cat)值分别为 0.08 mM 和 35 sec(-1)，最适 pH 和温度分别为 5.0 和 60°C。
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