Additional Carbohydrate-Binding Modules Enhance the Insoluble Substrate-Hydrolytic Activity of β-1,3-Glucanase from Alkaliphilic Nocardiopsis sp F96
Additional Carbohydrate-Binding Modules Enhance the Insoluble Substrate-Hydrolytic Activity of β-1,3-Glucanase from Alkaliphilic Nocardiopsis sp F96
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DOI：
10.1271/bbb.80846
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发表时间：
2009-05-01
影响因子：
1.6
通讯作者：
Nakamura, Satoshi
中科院分区：
文献类型：
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作者：
Koizumi, Naoya;Masuda, Sumiko;Nakamura, Satoshi
来自拟诺卡氏菌属物种F96的β-1，3-葡聚糖酶（BglF）仅由催化结构域组成。为了提高BglF的酶学性质，我们尝试构建BglF与环状芽孢杆菌（Bacillus circulans）IAM 1165的β-1，3-葡聚糖酶H的C-末端附加结构域（CAD）和N-末端附加结构域（NAD）以及嗜碱芽孢杆菌（Bacillus sp.）J813的几丁质酶的几丁质结合结构域（ChBD）的嵌合酶。构建并表征了CAD-融合的BglF（BglF-CAD）、NAD-融合的BglF（NAD-BglF）、NAD-和CAD-融合的BglF（NAD-BglF-CAD）和ChBD-融合的BglF（BglF-ChBD）。CAD的添加导致对不溶性β-1，3-葡聚糖的结合能力和水解活性增加。与BglF-CAD一样，BglF-CUD对茯苓多糖的结合能力和水解活性也有所增加。在pH 9-10范围内，BglF-CAD的水解活性高于BglF。BglF-CAD和BglF-ChBD在50-70 ℃左右的相对活性高于BglF。
beta-1,3-Glucanase (BglF) from Nocardiopsis sp. F96 is composed of only a catalytic domain. To improve the enzymatic properties of BglF, we attempted to construct chimeric enzymes consisting of BglF and some carbohydrate-binding modules, such as the C-terminal additional domain (CAD) and the N-terminal additional domain (NAD) of beta-1,3-glucanase H from Bacillus circulans IAM1165 and the chitin-binding domain (ChBD) of chitinase from alkaliphilic Bacillus sp. J813. CAD-fused BglF (BglF-CAD), NAD-fused BglF (NAD-BglF), both NAD- and CAD-fused BglF (NAD-BglF-CAD) and ChBD-fused BglF (BglF-ChBD) were constructed and characterized. The addition of CAD caused increases in binding abilities and hydrolytic activities toward insoluble beta-1,3-glucans. As well as BglF-CAD, the binding ability and hydrolytic activity of BglF-CUD toward pachyman were also increased. The hydrolytic activity of BglF-CAD at pH 9-10 was higher than that of BglF. The relative activities of BglF-CAD and BglF-ChBD at around 50-70 degrees C were higher than that of BglF.