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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
1993-7-19
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pubmed:abstractText |
The phenotypic features of strain GJ1B, an unidentified marine bacterium that degrades agar [Young, K. S. Bhattacharjee, S. S. & Yaphe, W. (1978) Carbohydr. Res. 66, 207-212], were investigated and its agarolytic system was characterized using 13C-NMR spectroscopy to analyse the agarose degradation products. The bacterium was assigned to the genus Alteromonas and the new combination A. agarlyticus (Cataldi) is proposed. An alpha-agarase, i.e. specific for the alpha(1-->3) linkages present in agarose, was purified to homogeneity from the culture supernatant by affinity chromatography on cross-linked agarose (Sepharose CL-6B) and by anion-exchange chromatography (Mono Q column). The major end product of agarose hydrolysis using the purified enzyme was agarotetraose. Using SDS/PAGE, the purified alpha-agarase was detected as a single band with a molecular mass of 180 kDa. After the affinity-chromatography step, however, the native molecular mass was approximately 360 kDa, suggesting that the native enzyme is a dimer which is dissociated to active subunits by anion-exchange chromatography. The isolectric point was estimated to be 5.3. Enzyme activity was observed using agar as the substrate over the pH range 6.0-9.0 with a maximum value at pH 7.2 in Mops or Tris buffer. The enzyme was inactivated by prolonged treatment at a pH below 6.5, or by temperatures over 45 degrees C or by removing calcium. In addition, a beta-galactosidase specific for the end products of the alpha-agarase was present in the alpha-agarase affinity-chromatography fraction, probably as part of a complex with this enzyme. The degradation of agarose by this agarase complex yielded a mixture of oligosaccharides in the agarotetraose series and the agarotriose series, the latter consisting of oligosaccharides with an odd number of galactose residues.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0014-2956
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
214
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
599-607
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pubmed:dateRevised |
2007-7-23
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pubmed:meshHeading |
pubmed-meshheading:8513809-Carbohydrate Conformation,
pubmed-meshheading:8513809-Chromatography, Affinity,
pubmed-meshheading:8513809-Chromatography, Ion Exchange,
pubmed-meshheading:8513809-Glycoside Hydrolases,
pubmed-meshheading:8513809-Gram-Negative Bacteria,
pubmed-meshheading:8513809-Hydrogen-Ion Concentration,
pubmed-meshheading:8513809-Hydrolysis,
pubmed-meshheading:8513809-Isoelectric Point,
pubmed-meshheading:8513809-Macromolecular Substances,
pubmed-meshheading:8513809-Magnetic Resonance Spectroscopy,
pubmed-meshheading:8513809-Molecular Weight,
pubmed-meshheading:8513809-Sepharose,
pubmed-meshheading:8513809-Substrate Specificity
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pubmed:year |
1993
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pubmed:articleTitle |
Purification and characterization of the alpha-agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B.
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pubmed:affiliation |
Centre d'Etudes d'Océanologie et de Biologie Marine, Centre National de la Recherche Scientifique, France.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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