Cloning and characterization of the glycogen branching enzyme gene existing in tandem with the glycogen debranching enzyme from Pectobacterium chrysanthemi PY35

Biochem Biophys Res Commun. 2003 Jan 3;300(1):93-101. doi: 10.1016/s0006-291x(02)02763-8.

Abstract

The glycogen branching enzyme gene (glgB) from Pectobacterium chrysanthemi PY35 was cloned, sequenced, and expressed in Escherichia coli. The glgB gene consisted of an open reading frame of 2196bp encoding a protein of 731 amino acids (calculated molecular weight of 83,859Da). The glgB gene is upstream of glgX and the ORF starts the ATG initiation codon and ends with the TGA stop codon at 2bp upstream of glgX. The enzyme was 43-69% sequence identical with other glycogen branching enzymes. The enzyme is the most similar to GlgB of E. coli and contained the four regions conserved among the alpha-amylase family. The glycogen branching enzyme (GlgB) was purified and the molecular weight of the enzyme was estimated to be 84kDa by SDS-PAGE. The glycogen branching enzyme was optimally active at pH 7 and 30 degrees C.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1,4-alpha-Glucan Branching Enzyme / chemistry
  • 1,4-alpha-Glucan Branching Enzyme / genetics*
  • 1,4-alpha-Glucan Branching Enzyme / metabolism
  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Conserved Sequence
  • DNA, Bacterial / genetics
  • Enterobacteriaceae / enzymology*
  • Enterobacteriaceae / genetics*
  • Escherichia coli / genetics
  • Genes, Bacterial*
  • Glycogen Debranching Enzyme System / genetics*
  • Molecular Sequence Data
  • Molecular Weight
  • Phylogeny
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Homology, Amino Acid

Substances

  • DNA, Bacterial
  • Glycogen Debranching Enzyme System
  • Recombinant Proteins
  • 1,4-alpha-Glucan Branching Enzyme