Gene expression of microbial gelatinase activity for porcine gelatine identification

Food Chem. 2021 Sep 1:355:129586. doi: 10.1016/j.foodchem.2021.129586. Epub 2021 Mar 13.

Abstract

In order to invent a porcine gelatine detection device using microbial resources, bacterial enzymes with a preference towards porcine gelatine and their candidate genes were evaluated. Five (n = 5) bacterial strains isolated from hot spring water and wet clay, Malaysia were screened for their gelatinase activity. The gelatinase enzyme was extracted and purified using ammonium sulphate precipitation prior to performing gelatinase assay on porcine, bovine and fish gelatine medium substrates. The G2 strain or Enterobacter aerogenes (Strain EA1) was selected for whole genome sequenced after showing a consistent trend of preference towards porcine gelatine. The gelatinase candidate gene gelEA1_9 was cloned and expressed. Based on one-way analysis of variance (ANOVA) with POST-HOC Duncan test (α = 0.05), the final product of gelEA1_9 was identified as a novel gelatinase. This gelatinase presented no significant difference in activity towards porcine gelatine. Hence, the present study demonstrated an enzyme-substrate interaction for porcine gelatine identification.

Keywords: Enterobacter aerogenes; Enzyme technology; Food authentication; Halal gelatine; Microbial resources.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cattle
  • Enterobacter aerogenes / enzymology
  • Enterobacter aerogenes / genetics
  • Fishes / metabolism
  • Gelatin / metabolism*
  • Gelatinases / chemistry
  • Gelatinases / genetics
  • Gelatinases / metabolism*
  • Gene Expression
  • RNA, Ribosomal, 16S / genetics
  • RNA, Ribosomal, 16S / metabolism
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Sequence Alignment
  • Substrate Specificity
  • Swine

Substances

  • Bacterial Proteins
  • RNA, Ribosomal, 16S
  • Recombinant Proteins
  • Gelatin
  • Gelatinases