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通过新型蛋白质工程策略提高 d-阿洛酮糖 3-差向异构酶的操作稳定性,以及从水果和蔬菜残渣中生产 d-阿洛酮糖。

Improved operational stability of d-psicose 3-epimerase by a novel protein engineering strategy, and d-psicose production from fruit and vegetable residues.

机构信息

Center of Innovative and Applied Bioprocessing (CIAB), Department of Biotechnology (DBT), Mohali, India.

Center of Innovative and Applied Bioprocessing (CIAB), Department of Biotechnology (DBT), Mohali, India.

出版信息

Bioresour Technol. 2016 Sep;216:121-7. doi: 10.1016/j.biortech.2016.05.053. Epub 2016 May 18.

Abstract

The aim of the present work was to improve stability of d-psicose 3-epimerase and biotransformation of fruit and vegetable residues for d-psicose production. The study established that N-terminal fusion of a yeast homolog of SUMO protein - Smt3 - can confer elevated optimal temperature and improved operational stability to d-psicose 3-epimerase. The Smt3-d-psicose 3-epimerase conjugate system exhibited relatively better catalytic efficiency, and improved productivity in terms of space-time yields of about 8.5kgL(-1)day(-1). It could serve as a promising catalytic tool for the pilot scale production of the functional sugar, d-psicose. Furthermore, a novel approach for economical production of d-psicose was developed by enzymatic and microbial bioprocessing of fruit and vegetable residues, aimed at epimerization of in situd-fructose to d-psicose. The bioprocessing led to achievement of d-psicose production to the extent of 25-35% conversion (w/w) of d-fructose contained in the sample.

摘要

本工作旨在提高 d-阿洛酮糖 3-差向异构酶的稳定性,并对水果和蔬菜残渣进行生物转化以生产 d-阿洛酮糖。研究表明,酵母同源物 SUMO 蛋白 Smt3 的 N 端融合可赋予 d-阿洛酮糖 3-差向异构酶更高的最适温度和改善的操作稳定性。Smt3-d-阿洛酮糖 3-差向异构酶缀合物系统表现出相对更好的催化效率,并在时空产率方面提高了约 8.5kgL(-1)day(-1)的生产力。它可以作为一种有前途的催化工具,用于功能糖 d-阿洛酮糖的中试生产。此外,还开发了一种通过水果和蔬菜残渣的酶和微生物生物加工来经济生产 d-阿洛酮糖的新方法,目的是将原位 d-果糖差向异构化为 d-阿洛酮糖。生物加工导致 d-阿洛酮糖的产量达到样品中所含 d-果糖的 25-35%转化率(w/w)。

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