Introduction of bifunctional alcohol/acetaldehyde dehydrogenase gene (adhE) in Fructobacillus fructosus settled its fructophilic characteristics

Res Microbiol. 2019 Jan-Feb;170(1):35-42. doi: 10.1016/j.resmic.2018.09.004. Epub 2018 Oct 4.

Abstract

Fructophilic lactic acid bacteria (FLAB) are unique in the sense that they prefer D-fructose over D-glucose as main carbon source. If D-glucose is metabolised, electron acceptors are required and significant levels of acetate are produced. These bacteria are found in environments rich in D-fructose, such as flowers, fruits and the gastrointestinal tract of insects feeding on fructose-rich diets. Fructobacillus spp. are representatives of this unique group, and their fructophilic characteristics are well conserved. In this study, the bifunctional alcohol/acetaldehyde dehydrogenase gene (adhE) from Leuconostoc mesenteroides NRIC 1541T was cloned into a plasmid and transferred to Fructobacillus fructosus NRIC 1058T. Differences in biochemical characteristics between the parental strain (NRIC 1058T) and the transformants were compared. Strain 1-11, transformed with the adhE gene, did not show any fructophilic characteristics, and the strain grew well on D-glucose without external electron acceptors. Accumulation of acetic acid, which was originally seen in the parental strain, was replaced with ethanol in the transformed strain. Furthermore, in silico analyses revealed that strain NRIC 1058T lacked the sugar transporters/permeases and enzymes required for conversion of metabolic intermediates. This may be the reason for poor carbohydrate metabolic properties recorded for FLAB.

Keywords: Alcohol/acetaldehyde dehydrogenase; Carbohydrate metabolism; Fructobacillus fructosus; Fructophilic lactic acid bacteria; Growth characteristics; adhE.

MeSH terms

  • Acetates / metabolism
  • Alcohol Dehydrogenase / chemistry
  • Alcohol Dehydrogenase / genetics*
  • Alcohol Dehydrogenase / metabolism*
  • Aldehyde Oxidoreductases / chemistry
  • Aldehyde Oxidoreductases / genetics
  • Aldehyde Oxidoreductases / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism*
  • Fructose / metabolism*
  • Gene Expression*
  • Glucose / metabolism
  • Leuconostoc / enzymology*
  • Leuconostoc / genetics
  • Leuconostocaceae / genetics*
  • Leuconostocaceae / growth & development
  • Leuconostocaceae / metabolism

Substances

  • Acetates
  • Bacterial Proteins
  • Fructose
  • Alcohol Dehydrogenase
  • Aldehyde Oxidoreductases
  • aldehyde dehydrogenase (NAD(P)+)
  • Glucose