Biotechnology and Bioprocess Engineering 2022; 27(2): 276-285  
Advanced Whole-cell Conversion for D-allulose Production Using an Engineered Corynebacterium glutamicum
Seong-Hee Jeong, Moonhyuk Kwon, and Seon-Won Kim
Seong-Hee Jeong, Moonhyuk Kwon*, Seon-Won Kim*
Division of Applied Life Science (BK21 Four), ABC-RLRC, PMBBRC, Gyeongsang National University, Jinju 52828, Korea
Tel: +82-55-772-1362; Fax: +82-55-759-9363
E-mail: mkwon@gnu.ac.kr
Tel: +82-55-772-1362; Fax: +82-55-759-9363
E-mail: swkim@gnu.ac.kr
Received: February 18, 2022; Revised: March 2, 2022; Accepted: March 3, 2022; Published online: April 30, 2022.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
D-allulose has received considerable attention as an alternative functional sugar for its zero caloric value with 70% relative sweetness compared to D-sucrose. Despite its strong potential as an alternative sweetener, recent industrial productions rely on a high-cost enzymatic method. Here, we advanced whole-cell conversion at high temperatures using Corynebacterium glutamicum expressing D-allulose 3-epimerase (DAE). By varying the reaction temperature from 25°C to 70°C, D-allulose conversion could reach the reaction equilibrium at high temperatures. The C. glutamicum showed superior reusability of cells at 60°C compared to Escherichia coli. We simplified the cell growth media and whole-cell conversion reaction solution. Clostridium hylemonae DAE (ChDAE) showed the highest thermostability and reusability among various DAE candidates. Finally, the ChDAE expression under the synthetic 2X-cT-T5 promoter could reduce the reaction time by 25%. Our result showed that 120 g/L of D-allulose can be produced from 400 g/L of D-fructose by reusable wholecell conversion at 55°C in 1.5 h. This study can be highly applicable in industrial economic production.
Keywords: Corynebacterium glutamicum, whole-cell conversion, D-allulose, D-allulose 3-epimerase, high temperature process

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