Expression and characterization of 1,4-α-glucan branching enzyme from Microvirga sp. MC18 and its application in the preparation of slowly digestible starch

Protein Expr Purif. 2021 Sep:185:105898. doi: 10.1016/j.pep.2021.105898. Epub 2021 May 5.

Abstract

Nutraceuticals containing modified starch with increased content of slowly-digestible starch (SDS) may reduce the prevalence of obesity, diabetes and cardiovascular diseases due to its slow digestion rate. Enzymatic methods for the preparation of modified starch have attracted increasing attention because of their low environmental impact, safety and specificity. In this study, the efficient glucan branching enzyme McGBE from Microvirga sp. MC18 was identified, and its relevant properties as well as its potential for industrial starch modification were evaluated. The purified McGBE exhibited the highest specificity for potato starch, with a maximal specific activity of 791.21 U/mg. A time-dependent increase in the content of α-1,6 linkages from 3.0 to 6.0% was observed in McGBE-modified potato starch. The proportion of shorter chains (degree of polymerization, DP < 13) increased from 29.2 to 63.29% after McGBE treatment, accompanied by a reduction of the medium length chains (DP 13-24) from 52.30 to 35.99% and longer chains (DP > 25) from 18.51 to 0.72%. The reduction of the storage modulus (G') and retrogradation enthalpy (ΔHr) of potato starch with increasing treatment time demonstrated that McGBE could inhibit the short- and long-term retrogradation of starch. Under the optimal conditions, the SDS content of McGBE-modified potato starch increased by 65.8% compared to native potato starch. These results suggest that McGBE has great application potential for the preparation of modified starch with higher SDS content that is resistant to retrogradation.

Keywords: Potato starch; Retrogradation; Slowly digestible starch; α-1,4-glucan branching enzyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1,4-alpha-Glucan Branching Enzyme / chemistry*
  • 1,4-alpha-Glucan Branching Enzyme / genetics
  • 1,4-alpha-Glucan Branching Enzyme / metabolism
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Cloning, Molecular
  • Dietary Supplements / analysis*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Genetic Vectors / chemistry
  • Genetic Vectors / metabolism
  • Humans
  • Hydrolysis
  • Kinetics
  • Methylobacteriaceae / chemistry
  • Methylobacteriaceae / enzymology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Starch / chemistry*
  • Substrate Specificity

Substances

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