Dual promoter strategy enhances co-expression of alpha-L-rhamnosidase and enhanced fluorescent protein for whole-cell catalysis and bioresource valorization

文献类型： 外文期刊

第一作者： Zhang, Fan

作者： Zhang, Fan;You, Shuai;Huang, Ting;Wang, Jin-Zheng;Zhu, Lin-Lin;Wang, Bo;Ye, Wang-Sheng;Herman, Richard Ansah;Luo, Heng;Wang, Jun;You, Shuai;Luo, Heng;Wang, Jun;You, Shuai;Luo, Heng;Wang, Jun;You, Shuai;Luo, Heng;Wang, Jun

作者机构：

关键词： Biocatalysis; Co-expression; Dual promoter; Enhanced fluorescent protein; alpha-L-rhamnosidase; Whole-cell catalysis

期刊名称：SCIENCE OF THE TOTAL ENVIRONMENT （ 影响因子：7.963； 五年影响因子：7.842 ）

ISSN： 0048-9697

年卷期： 2020 年 722 卷

页码：

收录情况： SCI

摘要： Developing circular economy is the only way to improve the efficiency of resource utilization. Whole-cell catalysis is an effective method to recycle enzymes, improve catalytic efficiency, and reduce production costs. The enzyme, alpha-L-rhamnosidase has considerable application prospects in the field of biocatalysis as it can hydrolyze a variety of alpha-L-rhamnoses. In the present study, the genes for alpha-L-rhamnosidase (rhaB1) and enhanced fluorescent protein (EGFP) were co-expressed using a bi-promoter expression vector pRSFDuet1 and their enzymatic properties were evaluated. To our knowledge, this study has established an effective rhamnosidase-fluorescent indicator and whole-cell catalytic system for the first time. Moreover, we analyzed the change in the activity of the crude rhaB1-EGFP as well as its whole-cell during the biocatalysis process using fluorescence intensity. Recombinant rhaB1-EGFP as a product which contains rhaB1 and EGFP showed higher thermal stability, pH stability, and conversion efficiency than rhaB1, and its optimumtemperature for rutin catalysis was ideal for industrial applications. Moreover, under the optimal conditions of a rutin concentration of 0.05 g/L, pH of 6.0, temperature of 40 degrees C, a yield of 92.5% was obtained. Furthermore, we demonstrated the relationship between the fluorescence intensity and enzyme activity. This study established a highly efficient whole-cell catalytic system whose activity can be evaluated by fluorescence intensity, providing a reference for enzyme recycling. (C) 2020 Elsevier B.V. All rights reserved.

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