Functional expression and enzymatic characterization of Lactobacillus plantarum cyclomaltodextrinase catalyzing novel acarbose hydrolysis

J Microbiol. 2018 Feb;56(2):113-118. doi: 10.1007/s12275-018-7551-3. Epub 2018 Feb 2.

Abstract

Cyclomaltodextrinases (CDases) belong to Glycoside Hydrolases (GH) family 13, which show versatile hydrolyzing and/or transglycosylation activity against cyclodextrin (CD), starch, and pullulan. Especially, some CDases have been reported to hydrolyze acarbose, a potent α-glucosidase inhibitor, and transfer the resulting acarviosine-glucose to various acceptors. In this study, a novel CDase (LPCD) gene was cloned from Lactobacillus plantarum WCFS1, which encodes 574 amino acids (64.6 kDa) and shares less than 44% of identities with the known CDase-family enzymes. Recombinant LPCD with C-terminal six-histidines was produced and purified from Escherichia coli. It showed the highest activity on β-CD at 45°C and pH 5.0, respectively. Gel permeation chromatography analysis revealed that LPCD exists as a dodecameric form (~826 kDa). Its hydrolyzing activity on β- CD is almost same as that on starch, whereas it can hardly attack pullulan. Most interestingly, LPCD catalyzed the unique modes of action in acarbose hydrolysis to produce maltose and acarviosine, as well as to glucose and acarviosineglucose.

Keywords: Lactobacillus plantarum; acarviosine; cyclomaltodextrinase; functional expression; novel acarbose hydrolysis.

MeSH terms

  • Acarbose / metabolism*
  • Amino Acid Sequence
  • Amino Sugars / metabolism
  • Cloning, Molecular
  • DNA, Bacterial / genetics
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial / genetics
  • Glucans / metabolism
  • Glucose / metabolism
  • Glycoside Hydrolases / classification
  • Glycoside Hydrolases / genetics*
  • Glycoside Hydrolases / metabolism*
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Lactobacillus plantarum / enzymology*
  • Lactobacillus plantarum / genetics*
  • Lactobacillus plantarum / metabolism
  • Maltose / metabolism
  • Molecular Weight
  • Substrate Specificity

Substances

  • Amino Sugars
  • DNA, Bacterial
  • Glucans
  • acarviosine
  • Maltose
  • pullulan
  • Glycoside Hydrolases
  • cyclomaltodextrinase
  • Glucose
  • Acarbose