Improving the thermostability of GH49 dextranase AoDex by site-directed mutagenesis

AMB Express

Original Article

Linxiang Xu¹  Nannan Liu¹  Zhen Wei¹  Jie Yang²  Shujun Wang²  Jinling Chen³ 

[1] Jiangsu Key Laboratory of Marine Bioresources and Environment, Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Jiangsu Ocean University, 222005, Lianyungang, China;Jiangsu Institute of Marine Resources Development, Jiangsu Ocean University, 222005, Lianyungang, China;Jiangsu Key Laboratory of Marine Bioresources and Environment, Co-Innovation Center of Jiangsu Marine Bio-Industry Technology, Jiangsu Ocean University, 222005, Lianyungang, China;School of Food Science and Engineering, Jiangsu Ocean University, 222005, Lianyungang, China;School of Food Science and Engineering, Jiangsu Ocean University, 222005, Lianyungang, China;

关键词: Dextranase;    Thermostability;    Arthrobacter oxydans;    Site-directed mutagenesis;    GH49;   

DOI  :  10.1186/s13568-023-01513-2

 received in 2022-12-31, accepted in 2023-01-08,  发布年份 2023

来源: Springer

【 摘 要 】

As an indispensable enzyme for the hydrolysis of dextran, dextranase has been widely used in the fields of food and medicine. It should be noted that the weak thermostability of dextranase has become a restricted factor for industrial applications. This study aims to improve the thermostability of dextranase AoDex in glycoside hydrolase (GH) family 49 that derived from Arthrobacter oxydans KQ11. Some mutants were predicted and constructed based on B-factor analysis, PoPMuSiC and HotMuSiC algorithms, and four mutants exhibited higher heat resistance. Compared with the wild-type, mutant S357P showed the best improved thermostability with a 5.4-fold increase of half-life at 60 °C, and a 2.1-fold increase of half-life at 65 °C. Furthermore, S357V displayed the most obvious increase in enzymatic activity and thermostability simultaneously. Structural modeling analysis indicated that the improved thermostability of mutants might be attributed to the introduction of proline and hydrophobic effects, which generated the rigid optimization of the structural conformation. These results illustrated that it was effective to improve the thermostability of dextranase AoDex by rational design and site-directed mutagenesis. The thermostable mutant of dextranase AoDex has potential application value, and it can also provide references for engineering other thermostable dextranases of the GH49 family.

【 授权许可】

CC BY   
© The Author(s) 2023

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