Recombinant expression and characterization of the glycogen branching enzyme from Vibrio vulnificus and its application in starch modification

Int J Biol Macromol. 2020 Jul 15:155:987-994. doi: 10.1016/j.ijbiomac.2019.11.062. Epub 2019 Nov 8.

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.

Keywords: Glycogen branching enzyme; In vitro digestibility; Resistant starch; Starch modification; Vibrio vulnificus.

MeSH terms

  • 1,4-alpha-Glucan Branching Enzyme / genetics
  • 1,4-alpha-Glucan Branching Enzyme / metabolism*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Digestion
  • Humans
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Starch / chemistry*
  • Vibrio vulnificus / enzymology*

Substances

  • Bacterial Proteins
  • Recombinant Proteins
  • Starch
  • 1,4-alpha-Glucan Branching Enzyme