Molecular Cloning and Expression of Two Novel [beta]-N-Acetylglucosaminidases from Silkworm Bombyx mori

β-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 β-N-acetylgluc...

Full description

Saved in:
Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2007-07, Vol.71 (7), p.1626
Main Authors: OKADA, Takahiro, ISHIYAMA, Seiji, SEZUTSU, Hideki, USAMI, Akihiro, TAMURA, Toshiki, MITA, Kazuei, FUJIYAMA, Kazuhito, SEKI, Tatsuji
Format: Article
Language:English
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:β-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 β-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 β-subunit homolog encoded by Drosophila melanogaster HEXO2 and the Spodoptera frugiperda β-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 α-3 and α-6 branches of a biantennary N-glycan substrate, and also hydrolyzed chitotriose to chitobiose.
ISSN:0916-8451
1347-6947