| Cloning of the gene of beta-N-acetylglucosaminidase from Lactobacillus casei ATCC 27092 and characterization of the enzyme expressed in Escherichia coli. | |
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MedLine Citation:
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PMID: 10823657 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The gene encoding beta-N-acetylglucosaminidase (GlcNAcase) of Lactobacillus casei ATCC 27092 was cloned and expressed in Escherichia coli. The gene consisted of 1581 nucleotides, and had a promoter, Shine-Dalgarno, and rho-independent type transcription termination sequences typical in bacteria. The protein deduced from the sequence consisted of 526 amino acids, and had a putative signal peptide of 14 amino acids and 5 possible asparagine-linked glycosylation sites. A conserved sequence was homologous to the 12 other hexosaminidases from different origins. The recombinant GlcNAcase (r-GlcNAcase) purified from the transformed E. coli had a MW of 39 kDa and lacked oligosaccharide chains. The isoelectric point and the optimum pH for the activity of r-GlcNAcase were similar to those of original GlcNAcases (o-GlcNAcase). However, the thermal stability was lower, and sensitivity to Cd2+, Fe2+, Cu2+ and sodium dodecyl sulfate (SDS) was higher than those of o-GlcNAcases, suggesting that the oligosaccharide moieties of the enzyme contribute to their stability. The Km value for p-nitrophenyl-N-acetyl-beta-1,4-D-glucosamine (PNP-GlcNAc) of r-GlcNAcase (6.4 microM) implied that the affinity of r-GlcNAcase for the substrate was 200-fold higher than that of the original ones. |
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Authors:
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M Senba; N Kashige; Y Nakashima; F Miake; K Watanabe |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Biological & pharmaceutical bulletin Volume: 23 ISSN: 0918-6158 ISO Abbreviation: Biol. Pharm. Bull. Publication Date: 2000 May |
Date Detail:
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Created Date: 2000-07-24 Completed Date: 2000-07-24 Revised Date: 2007-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9311984 Medline TA: Biol Pharm Bull Country: JAPAN |
Other Details:
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Languages: eng Pagination: 527-31 Citation Subset: IM |
Affiliation:
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Faculty of Pharmaceutical Sciences, Fukuoka University, Japan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acetylglucosaminidase
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biosynthesis,
chemistry,
genetics* Amino Acid Sequence Base Sequence Cloning, Molecular DNA Escherichia coli Hexosaminidases / chemistry Lactobacillus casei / enzymology, genetics* Molecular Sequence Data Recombinant Proteins / biosynthesis, chemistry, genetics Sequence Homology, Amino Acid |
| Chemical | |
Reg. No./Substance:
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0/Recombinant Proteins; 9007-49-2/DNA; EC 3.2.1.-/Hexosaminidases; EC 3.2.1.52/Acetylglucosaminidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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