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Characterization of a β-D-mannosidase from a marine gastropod, Aplysia kurodai.
MedLine Citation:
PMID:  22366551     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A β-D-mannosidase (EC 3.2.1.25) with a molecular mass of approximately 100kDa was purified from the digestive fluid of a marine gastropod Aplysia kurodai by ammonium sulfate fractionation followed by column chromatographies on TOYOPEARL Butyl-650M, TOYOPEARL DEAE-650M, and Superdex 200 10/300 GL. This enzyme, named AkMnsd in the present study, showed optimal activities at pH 4.5 and 40°C and was stable at the acidic pH range from 2.0 to 6.7 and the temperature below 38°C. The Km and Vmax values for AkMnsd determined at pH 6.0 and 30°C with p-nitrophenyl β-d-mannopyranoside were 0.10mM and 3.75μmol/min/mg, respectively. AkMnsd degraded various polymer mannans as well as mannooligosaccharides liberating mannose as a major degradation product. Linear mannan from green alga Codium fragile was completely depolymerized by AkMnsd in the presence of AkMan, an endolytic β-mannanase, which we previously isolated from the same animal (Zahura et al., Comp. Biochem. Physiol. B 157, 137-148 (2010)). A cDNA encoding AkMnsd was amplified from the Aplysia hepatopancreas cDNA by the PCR using degenerated primers designed on the basis of N-terminal and internal amino-acid sequences of AkMnsd. The cloned AkMnsd cDNA consisted of 2985bp and encoded an amino-acid sequence of 931 residues with the calculated molecular mass of 101,970Da. The deduced sequence of AkMnsd showed 20-43% amino-acid identity to those of glycoside-hydrolase-family 2 (GHF2) β-mannosidases. The catalytically important amino-acid residues determined in GHF2 enzymes were completely conserved in AkMnsd. Thus, AkMnsd is regarded as a new member of GHF2 mannosidase from marine gastropod.
Authors:
Umme Afsari Zahura; Mohammad Matiur Rahman; Akira Inoue; Takao Ojima
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-2-18
Journal Detail:
Title:  Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology     Volume:  -     ISSN:  1879-1107     ISO Abbreviation:  -     Publication Date:  2012 Feb 
Date Detail:
Created Date:  2012-2-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9516061     Medline TA:  Comp Biochem Physiol B Biochem Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012. Published by Elsevier Inc.
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