Biochemical Characterization of Alkaliphilic Cyclodextran Glucanotransferase from an Alkaliphilic Bacterium, Paenibacillus daejeonensis

J Microbiol Biotechnol. 2018 Dec 28;28(12):2029-2035. doi: 10.4014/jmb.1810.10007.

Abstract

Cycloisomaltooligosaccharide glucanotransferase (CITase) was isolated from alkaliphilic Paenibacillus daejeonensis via an amino acid homology search for the reported CITase. The recombinant alkaliphilic CITase (PDCITase) from P. daejeonensis was expressed in an Escherichia coli expression system and purified as a single protein band of 111 kDa. PDCITase showed optimum activity at pH 8.0 and retained 100% of activity within a broad pH range (7.0-11.5) after 18 h, indicating alkaliphilic or alkalistable CITase properties. In addition, PDCITase produced CI-7 to CI-17, CI-18, and CI-19, which are relatively large cycloisomaltooligosaccharides yet to be reported. Therefore, these large cycloisomaltooligosaccharides can be applied to the improvement of water solubility of pharmaceutical biomaterials.

Keywords: Cyclodextran; Paenibacillus daejeonensis; cycloisomaltooligosaccharide glucanotransferase; glycoside hydrolase family 66.

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • Enzyme Assays
  • Enzyme Stability
  • Escherichia coli / genetics
  • Gene Expression Regulation, Bacterial
  • Glucosyltransferases / chemistry*
  • Glucosyltransferases / genetics*
  • Glucosyltransferases / isolation & purification*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Metals
  • Paenibacillus / enzymology*
  • Paenibacillus / genetics*
  • Recombinant Proteins / genetics
  • Solubility
  • Substrate Specificity
  • Temperature

Substances

  • Metals
  • Recombinant Proteins
  • Glucosyltransferases
  • cycloisomaltooligosaccharide glucanotransferase