Characterization of endo-β-N-acetylglucosaminidase from alkaliphilic Bacillus halodurans C-125

Kiyotaka Fujita, Hideto Takami, Kenji Yamamoto, Kaoru Takegawa

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The genome sequencing project on alkaliphilic Bacillus halodurans C-125 revealed a putative endo-β-N-acetylglucosaminidase (Endo-BH), which consists of a signal peptide of 24 amino acids, a catalytic region of 634 amino acids exhibiting 50.1% identity with the endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae (Endo-A), and a C-terminal tail of 220 amino acids. Transformed Escherichia coli cells carrying the Endo-BH gene exhibited endo-β-N-acetylglucosaminidase activity. Recombinant Endo-BH hydrolyzed high-mannose type oligosaccharides and hybrid type oligosaccharides, and showed transglycosylation activity. On deletion of 219 C-terminal amino acid residues of Endo-BH, the wild type level of activity was retained, whereas with deletions of the Endo-A homolog domain, the proteins were expressed as inclusion bodies and these activities were reduced. These results suggest that the enzymatic properties of Endo-BH are similar to those of Endo-A, and that the C-terminal tail does not affect the enzyme activity. Although the C-terminal tail region is not essential for enzyme activity, the sequence is also conserved among endo-β-N-acetylglucosaminidases of various origins.

Original languageEnglish
Pages (from-to)1059-1066
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume68
Issue number5
DOIs
Publication statusPublished - May 1 2004

Fingerprint

Acetylglucosaminidase
Bacilli
Bacillus
Amino Acids
Enzyme activity
Oligosaccharides
Genes
Arthrobacter
Inclusion Bodies
Enzymes
Mannose
Protein Sorting Signals
Escherichia coli
Catalytic Domain
Genome
endo-alpha-sialidase
Proteins

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

Cite this

Characterization of endo-β-N-acetylglucosaminidase from alkaliphilic Bacillus halodurans C-125. / Fujita, Kiyotaka; Takami, Hideto; Yamamoto, Kenji; Takegawa, Kaoru.

In: Bioscience, Biotechnology and Biochemistry, Vol. 68, No. 5, 01.05.2004, p. 1059-1066.

Research output: Contribution to journalArticle

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