Molecular cloning and expression of two novel beta-N-acetylglucosaminidases from silkworm Bombyx mori.
Biosci Biotechnol Biochem
; 71(7): 1626-35, 2007 Jul.
Article
em En
| MEDLINE
| ID: mdl-17617724
ABSTRACT
Beta-N-acetylglucosaminidase is a major glycosidase involved in several physiological processes, such as fertilization, metamorphosis, glycoconjugate degradation, and glycoprotein biosynthesis in insects. A search using the Bombyx mori cDNA database revealed the existence of two putative beta-N-acetylglucosaminidase genes. Their full-length cDNAs were cloned by rapid amplification of cDNA ends and polymerase chain reaction using specific primers, and named BmGlcNAcase1 and BmGlcNAcase2. A BLAST search revealed that BmGlcNAcase1 and BmGlcNAcase2 are homologous to a beta-subunit homolog encoded by Drosophila melanogaster HEXO2 and the Spodoptera frugiperda beta-N-acetylglucosaminidase gene respectively. The recombinant proteins of BmGlcNAcase1 and BmGlcNAcase2 without putative transmembrane domains were expressed in the yeast Pichia pastoris. Both enzymes showed broad substrate specificity, and cleaved terminal N-acetylglucosamine residues from the alpha-3 and alpha-6 branches of a biantennary N-glycan substrate, and also hydrolyzed chitotriose to chitobiose.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Acetilglucosaminidase
/
Bombyx
/
Clonagem Molecular
Limite:
Animals
Idioma:
En
Revista:
Biosci Biotechnol Biochem
Ano de publicação:
2007
Tipo de documento:
Article