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Recombinant expression and characterization of the glycogen branching enzyme from Vibrio vulnificus and its
Lingling Li1, Lingqia Su1, Fan Hu1
1State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; School of Biotechnology, Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China; International Joint Laboratory on Food Safety, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.
Abstract:
Resistant starch (RS) is helpful in controlling and preventing metabolic syndrome relevant diseases. However, the RS content of natural starch and modified starch produced by enzymatic method is generally low. To solve this problem, we selected the glycogen branching enzyme from Vibrio vulnificus (VvGBE) and investigated its application. Firstly, it was expressed in E. coli with the enzyme activity was 53.33 U/mL, and its optimum temperature and pH was 35 °C and 7.5, respectively. The half-life of VvGBE at 35 °C was 10 h, and the enzyme was most stable at pH 9.5. When we used the recombinant enzyme to treat corn starch, the content of RS increased by 19.41%, which was higher than that achieved with other enzymes. More specially, the conversion of slowly digestible starch to RS, which was only demonstrated in chemical modification, was accomplished. The fine structure of the modified starch was further investigated. Results showed that the number of short chains (DP < 13) increased to 90.58%, and the α-1,6 linkages ratio increased from 7.19% to 15.64%. The increase of short chains and α-1,6 linkages may contribute to high RS content. This study can provide a reference for the development of modified starch with lower digestibility.
