Thermostability Improvement of the d-Allulose 3-Epimerase from Dorea sp. CAG317 by Site-Directed Mutagenesis at the Interface Regions

Journal of Agricultural and Food Chemistry
Wenli ZhangWanmeng Mu

Abstract

d-Allulose is a low-calorie sweetener and has broad applications in the food, cosmetics, and pharmaceutical industries. Recently, most studies focus on d-allulose production from d-fructose by d-allulose 3-epimerase (DAEase). However, the major blocker of industrial production of d-allulose is the poor thermostability. In this study, site-directed mutagenesis at the interface regions of Dorea sp. DAEase was carried out, and the F154Y/E191D/I193F mutation was obtained. The mutant protein displayed much higher thermostability, with a t1/2 value of 20.47 h (50 °C) and a Tm value of 74.18 °C. Compared with the wild-type DAEase, the t1/2 value at 50 °C increased by 5.4-fold, and the Tm value increased by 17.54 °C. In the d-allulose production from 500 g/L d-fructose, 148.2 g/L d-allulose could be obtained by F154Y/E191D/I193F mutant protein. The results suggest that site-directed mutagenesis at the interface regions is an efficient approach for improving the thermostability of DAEase.

References

Mar 1, 1993·Trends in Biotechnology·V V Mozhaev
Jan 1, 1997·Methods in Enzymology·D EisenbergJ U Bowie
Nov 16, 2001·Asia Pacific Journal of Clinical Nutrition·T MatsuoH Suzuki
Nov 23, 2005·Bioinformatics·Konstantin ArnoldTorsten Schwede
Dec 31, 2005·Journal of Bioscience and Bioengineering·Maki K TakataMasaaki Tokuda
Feb 8, 2006·Applied and Environmental Microbiology·Hye-Jung KimDeok-Kun Oh
Jun 15, 2011·Journal of Agricultural and Food Chemistry·Wanmeng MuBo Jiang
May 10, 2012·Applied Microbiology and Biotechnology·Wanmeng MuLeon Zhou
Nov 10, 2013·Journal of Agricultural and Food Chemistry·Wenli ZhangWanmeng Mu
Dec 7, 2013·Acta Crystallographica. Section D, Biological Crystallography·Keiko UechiGoro Takata
Apr 5, 2014·Natural Product Reports·Madhu Emmadi, Suvarn S Kulkarni
Apr 29, 2014·Journal of Basic Microbiology·Wenjia HanYuanxia Sun
Oct 8, 2016·Journal of Bioscience and Bioengineering·Akihide YoshiharaPushpa Kiran Gullapalli
Jan 24, 2017·Biotechnology Advances·Wenli ZhangWanmeng Mu
Apr 19, 2017·Journal of the Science of Food and Agriculture·Masahiro OgawaMakiko Ogawa
Sep 25, 2017·Nutrition·Tomonori KimuraKazuhiro Okuma
Nov 3, 2017·Carbohydrate Research·Aimin LiZijie Li

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Citations

Mar 3, 2020·Frontiers in Bioengineering and Biotechnology·Suwei JiangJingjing Liu
Apr 11, 2021·FEBS Open Bio·Hiromi YoshidaShigehiro Kamitori
Dec 31, 2021·Critical Reviews in Food Science and Nutrition·Wenli ZhangWanmeng Mu

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