Molecular cloning and characterization of an endo-1,3-beta-D-glucanase from the mollusk Spisula sachalinensis

Comp Biochem Physiol B Biochem Mol Biol. 2004 Feb;137(2):169-78. doi: 10.1016/j.cbpc.2003.10.018.

Abstract

cDNA encoding the endo-1,3-beta-d-glucanase from Spisula sachalinensis (LIV) was amplified by PCR using oligonucleotides deduced from the N-terminal end peptide sequence. Predicted enzyme structure consists of 444 amino acids with a signal sequence. The mature enzyme has 316 amino acids and its deduced amino acid sequence coincides completely with the N-terminal end (38 amino acids) of the beta-1,3-glucanase (LIV) isolated from the mollusk. The enzyme sequence from Val 121 to Met 441 reveals closest homology with Pacifastacus leniusculus lipopolysaccharide- and beta-1,3-glucan-binding protein and with coelomic cytolytic factors from Lumbricus terrestris. The mollusk glucanase also shows 36% identity and 56% similarity with beta-1,3-glucanase of the sea urchin Strongylocentrotus purpuratus. It is generally considered that invertebrate glucanase-like proteins containing the bacterial glucanase motif have evolved from an ancient beta-1,3-glucanase gene, but most of them lost their glucanase activity in the course of evolution and retained only the glucan-binding activity. A more detailed evaluation of the protein folding elicited very interesting relationships between the active site of LIV and other enzymes, which hydrolyze native glucans.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Astacoidea / genetics
  • Cloning, Molecular
  • DNA, Complementary
  • Glucan Endo-1,3-beta-D-Glucosidase / genetics*
  • Molecular Sequence Data
  • Mollusca / genetics*
  • Phylogeny
  • Protein Structure, Tertiary
  • Sea Urchins / genetics
  • Sequence Alignment
  • Sequence Homology, Amino Acid

Substances

  • DNA, Complementary
  • Glucan Endo-1,3-beta-D-Glucosidase