Engineering the thermostability of d-lyxose isomerase from Caldanaerobius polysaccharolyticus via multiple computer-aided rational design for efficient synthesis of d-mannose

Hao Wu; Ming Yi; Xiaoyi Wu; Yating Ding; Minghui Pu; Li Wen; Yunhui Cheng; Wenli Zhang; Wanmeng Mu

Synthetic and Systems Biotechnology (Jun 2023)

Engineering the thermostability of d-lyxose isomerase from Caldanaerobius polysaccharolyticus via multiple computer-aided rational design for efficient synthesis of d-mannose

Hao Wu,
Ming Yi,
Xiaoyi Wu,
Yating Ding,
Minghui Pu,
Li Wen,
Yunhui Cheng,
Wenli Zhang,
Wanmeng Mu

Affiliations

Hao Wu: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China; State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China
Ming Yi: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China
Xiaoyi Wu: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China
Yating Ding: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China
Minghui Pu: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China
Li Wen: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China
Yunhui Cheng: School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, 410114, China
Wenli Zhang: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China; Corresponding author. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
Wanmeng Mu: State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

Journal volume & issue: Vol. 8, no. 2
pp. 323 – 330

Abstract

Read online

d-Mannose is an attractive functional sugar that exhibits many physiological benefits on human health. The demand for low-calorie sugars and sweeteners in foods are increasingly available on the market. Some sugar isomerases, such as d-lyxose isomerase (d-LIase), can achieve an isomerization reaction between d-mannose and d-fructose. However, the weak thermostability of d-LIase limits its efficient conversion from d-fructose to d-mannose. Nonetheless, few studies are available that have investigated the molecular modification of d-LIase to improve its thermal stability. In this study, computer-aided tools including FireProt, PROSS, and Consensus Finder were employed to jointly design d-LIase mutants with improved thermostability for the first time. Finally, the obtained five-point mutant M5 (N21G/E78P/V58Y/C119Y/K170P) showed high thermal stability and catalytic activity. The half-life of M5 at 65 °C was 10.22 fold, and the catalytic efficiency towards 600 g/L of d-fructose was 2.6 times to that of the wild type enzyme, respectively. Molecular dynamics simulation and intramolecular forces analysis revealed a thermostability mechanism of highly rigidity conformation, newly formed hydrogen bonds and π-cation interaction between and within protein domains, and redistributed surface electrostatic charges for the mutant M5. This research provided a promising d-LIase mutant for the industrial production of d-mannose from d-fructose.

Published in Synthetic and Systems Biotechnology

ISSN: 2405-805X (Online)
Publisher: KeAi Communications Co., Ltd.
Country of publisher: China
LCC subjects: Technology: Chemical technology: Biotechnology; Science: Biology (General)
Website: https://www.keaipublishing.com/en/journals/synthetic-and-systems-biotechnology/

About the journal

Abstract

Keywords