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Expression, characterization and mutagenesis of the gene encoding β-N-acetylglucosaminidase from Aeromonas caviae CB101

Paper ID Volume ID Publish Year Pages File Format Full-Text
18393 42721 2006 7 PDF Available
Title
Expression, characterization and mutagenesis of the gene encoding β-N-acetylglucosaminidase from Aeromonas caviae CB101
Abstract

The gene encoding β-N-acetylglucosaminidase (nagA1) from Aeromonas caviae CB101 was cloned, and its nucleotide sequence was determined. The open reading frame of the gene consisted of 2661 bp encoding a polypeptide of 887 amino acids including a putative signal peptide of 22 amino acids. The deduced amino acid sequence of nagA1 showed high similarities with β-N-acetylglucosaminidase from various organisms, belonging to family 20 of glycosyl hydrolases. The nagA1 gene was expressed in Escherichia coli. The purified enzyme hydrolyzed N-acetylchitooligomers from N,N′-diacetylchitobiose to hexa-N-acetylchitohexaose with highest activity towards N,N′-diacetylchitobiose. The enzyme could also rapidly cleave p-nitrophenyl-N-acetyl-β-d-glucosaminide and p-nitrophenyl-N-acetyl-β-d-galactosaminide. The hydrolytic activity of NagA1 was optimal at 50 °C and pH 8.0. Substitution of Glu538, Asp537 with Ala caused abolishing or largely decreasing of the enzyme activity, respectively, indicated that the predicted active sites Glu538, Asp537 were essential to the enzymatic activity of NagA1.

Keywords
Aeromonas caviae; β-N-Acetylglucosaminidase; Site-directed mutagenesis
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Expression, characterization and mutagenesis of the gene encoding β-N-acetylglucosaminidase from Aeromonas caviae CB101
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 38, Issue 6, 1 April 2006, Pages 765–771
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us