Structure of the Clostridium stercorarium gene celY encoding the exo-1,4-beta-glucanase Avicelase II.
The nucleotide sequence of the celY gene coding for the thermostable exo-1,4-beta-glucanase Avicelase II of Clostridium stercorarium was determined. The gene consists of an ORF of 274Z bp which encodes a preprotein of 914 amino acids with a molecular mass of 103 kDa. The signal-peptide cleavage site was identified by comparison with the N-terminal amino acid sequence of Avicelase II purified from C stercorarium. The celY gene is located in close vicinity to the celZ gene coding for the endo-1,4-beta-glucanase Avicelase I. The CelY-encoding sequence was isolated from genomic DNA of C. stercorarium with the PCR technique. The recombinant enzyme produced in Escherichia coli as a LacZ'-CelY fusion protein could be purified using a simple two-step procedure. The properties of CelY proved to be consistent with those of Avicelase II purified from C. stercorarium. Sequence comparison revealed that CelY consists of an N-terminal catalytic domain flanked by a domain of 95 amino acids with unknown function joined to a type III cellulose-binding domain. The catalytic domain belongs to the recently proposed family L of cellulases (family 48 of glycosyl hydrolases).
Bronnenmeier
Karin
K
Institute for Microbiology, Technical University Munich, Arcisstra�e 21, D-80290 M�nchen, Federal Republic of Germany.
Kundt
Kerstin
K
Institute for Microbiology, Technical University Munich, Arcisstra�e 21, D-80290 M�nchen, Federal Republic of Germany.
Riedel
Kathrin
K
Institute for Microbiology, Technical University Munich, Arcisstra�e 21, D-80290 M�nchen, Federal Republic of Germany.
Schwarz
Wolfgang H
WH
Institute for Microbiology, Technical University Munich, Arcisstra�e 21, D-80290 M�nchen, Federal Republic of Germany.
Staudenbauer
Walter L
WL
Institute for Microbiology, Technical University Munich, Arcisstra�e 21, D-80290 M�nchen, Federal Republic of Germany.
D000595:Amino Acid Sequence; D001483:Base Sequence; D002480:Cellulase / Q000235:genetics*; D043366:Cellulose 1,4-beta-Cellobiosidase; D003001:Cloning, Molecular; D003013:Clostridium / Q000235:genetics*; D005798:Genes, Bacterial; D044643:Glucan 1,4-beta-Glucosidase; D008969:Molecular Sequence Data; D017421:Sequence Analysis; D001617:beta-Glucosidase / Q000235:genetics*
D001617:beta-Glucosidase; D002480:Cellulase; D044643:Glucan 1,4-beta-Glucosidase; D043366:Cellulose 1,4-beta-Cellobiosidase