Changes in the activity of the multifunctional beta-glycosyl hydrolase (Cel44C-Man26A) from Paenibacillus polymyxa by removal of the C-terminal region to minimum size

Biotechnol Lett. 2008 Jun;30(6):1061-8. doi: 10.1007/s10529-008-9640-6. Epub 2008 Jan 26.

Abstract

Paenibacillus polymyxa GS01 secretes Cel44C-Man26A as a multifunctional enzyme with cellulase, xylanase, lichenase, and mannanase activities. Cel44C-Man26A consists of 1,352 amino acids in which present a catalytic domain (CD) of the glycosyl hydrolase family 44 (GH44), fibronectin domain type 3 (Fn3), catalytic domain of glycosyl hydrolase family 26 (GH26), and a cellulose-binding module type 3 (CBM3). A truncated Cel44C-Man26A protein, consisting of 549 amino acid residues, reacted as a multifunctional mature enzyme despite the absence of the 10 amino acids containing GH44, Fn3, GH26, and CBM3. However, the multifunctional activity was not found in the mature Cel44C-Man26A protein truncated to less than 548 amino acids. The truncated Cel44C-Man26A proteins showed the optimum pH for the lichenase activity was pH 7.0, pH 6.0 for the xylanase and mannanase, and pH 5.0 for the cellulase. The truncated Cel44C-Man26A proteins exhibited enzymatic activity 40-120% higher than the full-length Cel44C.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Artificial Gene Fusion
  • Bacteria / enzymology*
  • Bacteria / genetics
  • Bacteria / metabolism
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / metabolism*
  • Cellulases / metabolism
  • Cloning, Molecular
  • Codon, Nonsense / genetics
  • DNA, Recombinant / biosynthesis
  • Electrophoresis, Polyacrylamide Gel
  • Endo-1,4-beta Xylanases / metabolism
  • Glycoside Hydrolases / biosynthesis
  • Glycoside Hydrolases / metabolism*
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Mutagenesis, Insertional
  • beta-Mannosidase / metabolism

Substances

  • Bacterial Proteins
  • Codon, Nonsense
  • DNA, Recombinant
  • Cellulases
  • Glycoside Hydrolases
  • beta-Mannosidase
  • licheninase
  • Endo-1,4-beta Xylanases