β-Glucosidase from Streptomyces griseus: Nanoparticle immobilisation and application to alkyl glucoside. synthesis
β-Glucosidase from Streptomyces griseus: Nanoparticle immobilisation and application to alkyl glucoside. synthesis
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DOI：
10.1016/j.pep.2017.01.011
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发表时间：
2017-04-01
影响因子：
1.6
通讯作者：
Henehan, G. T. M.
中科院分区：
文献类型：
--
作者：
Kumar, P.;Ryan, B.;Henehan, G. T. M.
从灰色链霉菌（Streptomycesgriseus）中克隆了一种新的β-葡萄糖苷酶，并在E.杆菌纯化的β-葡糖苷酶（44 kDa）在37 ℃、pH 7.2下以对硝基苯基-β-D吡喃葡萄糖苷为底物时的Km为8.6 +/- 0.5 mM，Vmax为217 +/- 5.0 μ moles（-1）min（-1）mg。酶的特点是在最适pH值（pH 6.9），最适温度（69 ℃）和溶剂和效应的影响。纯化S.通过简单的吸附，在没有共价修饰的情况下，将灰色杆菌β-葡萄糖苷酶成功地固定到氧化锌（ZnO）纳米颗粒上。即使在大量洗涤后，它仍然保持紧密结合，并且可以重复使用多达十次而不会显着损失活性。固定化酶的最适温度和热稳定性均高于游离酶。在固定形式的酶容易催化烷基葡糖苷的合成。(C)2017爱思唯尔公司All rights reserved.
A novel beta-glucosidase from Streptomyces griseus was cloned and overexpressed in E. coli. The purified beta-glucosidase (44 kDa) had a K-m of 8.6 +/- 0.5 mM and a V-max of 217 +/- 5.0 mu moles(-1)min(-1)mg at 37 C-omicron, pH 7.2 with p-nitrophenyl-beta-D glucopyranoside as substrate. The enzyme was characterised in terms of pH optimum (pH 6.9), temperature optimum (69 C-omicron) and the influence of solvents and effectors. Purified S. griseus beta-glucosidase was successfully immobilised, by simple absorption, onto zinc oxide (ZnO) nanoparticles without covalent modification. It remained tightly bound even after extensive washing and could be reused up to ten times without significant loss of activity. The immobilised enzyme had a higher optimum temperature and greater thermostability than the free enzyme. In immobilised form the enzyme readily catalysed the synthesis of alkyl glucosides. (C) 2017 Elsevier Inc. All rights reserved.