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从产黄青霉中克隆、纯化和研究重组 GH3 家族β-葡萄糖苷酶。

Cloning, purification and study of recombinant GH3 family β-glucosidase from Penicillium verruculosum.

机构信息

Federal Research Centre "Fundamentals of Biotechnology" of the Russian Academy of Sciences, Leninsky Pr. 33/2, Moscow, 119071, Russia.

Federal Research Centre "Fundamentals of Biotechnology" of the Russian Academy of Sciences, Leninsky Pr. 33/2, Moscow, 119071, Russia.

出版信息

Biochimie. 2020 Jan;168:231-240. doi: 10.1016/j.biochi.2019.11.009. Epub 2019 Nov 19.

Abstract

A novel bgl1 gene, encoding GH3 family β-glucosidase from Penicillium verruculosum (PvBGL), was cloned and heterologously expressed in P. canescens RN3-11-7 (niaD-) strain under the control of the strong xylA gene promoter. The recombinant rPvBGL was purified and their properties were studied in comparison with those of rAnBGL from Aspergillus niger expressed previously in the same fungal host. The rPvBGL had an observed molecular mass of 90 kDa (SDS-PAGE data) and displayed the enzyme maximum activity at pH 4.6 and 65 °C. The enzyme half-life time at 60 °C was found to be 87 min. Unlike the rAnBGL, the rPvBGL was not adsorbed on microcrystalline cellulose, which gives the latter enzyme an advantage in cellulose conversion with a longer time of hydrolysis.

摘要

从疣孢青霉(PvBGL)中克隆并异源表达了一种新型的 bgl1 基因,该基因编码 GH3 家族β-葡萄糖苷酶。在强 xylA 基因启动子的控制下,该基因在 P. canescens RN3-11-7(niaD-)菌株中进行表达。重组 rPvBGL 被纯化,并与先前在相同真菌宿主中表达的来自黑曲霉的 rAnBGL 的性质进行了比较。rPvBGL 的观察到的分子量为 90 kDa(SDS-PAGE 数据),并在 pH 4.6 和 65°C 时显示出最大酶活性。在 60°C 时,酶的半衰期为 87 分钟。与 rAnBGL 不同,rPvBGL 不被微晶纤维素吸附,这使得后者在水解时间更长的纤维素转化中具有优势。

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