DC Field | Value | Language |
---|---|---|
dc.contributor.author | Park, Da Yeon | ko |
dc.contributor.author | Chi, Won-Jae | ko |
dc.contributor.author | Park, Jae-Seon | ko |
dc.contributor.author | Chang, Yong-Keun | ko |
dc.contributor.author | Hong, Soon-Kwang | ko |
dc.date.accessioned | 2015-04-08T02:26:31Z | - |
dc.date.available | 2015-04-08T02:26:31Z | - |
dc.date.created | 2015-03-03 | - |
dc.date.created | 2015-03-03 | - |
dc.date.created | 2015-03-03 | - |
dc.date.issued | 2015-01 | - |
dc.identifier.citation | APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, v.175, no.2, pp.733 - 747 | - |
dc.identifier.issn | 0273-2289 | - |
dc.identifier.uri | http://hdl.handle.net/10203/195455 | - |
dc.description.abstract | An agarase gene (agaH71) was identified from Pseudoalteromonas hodoensis, an agar utilizing marine bacterium. The nucleotide sequence revealed that AgaH71 had significant homology to glycosyl hydrolase (GH) 16 agarases. agaH71 encodes a primary translation product (32.7 kDa) of 290 amino acids, including a 21-amino-acid signal peptide. The entire AgaH71 was expressed in a fused protein with glutathione-S-transferase (GST) at its N-terminal (GST-AgaH71) in Escherichia coli. Purified GST-AgaH71 had an apparent molecular weight of 59 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which was consistent with the calculated molecular weight (58.7 kDa). Agarase activity of the purified protein was confirmed by zymogram assay. GST-AgaH71 could hydrolyze p-nitrophenyl-beta-d-galactopyranoside, but not p-nitrophenyl-alpha-d-galactopyranoside. The optimum pH and temperature for GST-AgaH71 were 6.0 and 45 A degrees C, respectively. GST-AgaH71 retained more than 95 and 90 % of its initial activity at 40 and 45 A degrees C after heat treatment for 60 min, respectively. The K (m) and V (max) for agarose were 28.33 mg/ml and 88.25 U/mg, respectively. GST-AgaH71 did not require metal ions for its activity, but severe inhibition by divalent metal ions was observed. Thin-layer chromatography (TLC) analysis, mass spectrometry, and nuclear magnetic resonance (NMR) spectrometry of the GST-AgaH71 hydrolysis products revealed that GST-AgaH71 is an endo-type beta-agarase that hydrolyzes agarose into predominantly neoagarotetraose and small proportions of neoagarobiose and neoagarohexaose. | - |
dc.language | English | - |
dc.publisher | HUMANA PRESS INC | - |
dc.title | Cloning, Expression, and Biochemical Characterization of a GH16 beta-Agarase AgaH71 from Pseudoalteromonas hodoensis H7 | - |
dc.type | Article | - |
dc.identifier.wosid | 000348103100010 | - |
dc.identifier.scopusid | 2-s2.0-84921033035 | - |
dc.type.rims | ART | - |
dc.citation.volume | 175 | - |
dc.citation.issue | 2 | - |
dc.citation.beginningpage | 733 | - |
dc.citation.endingpage | 747 | - |
dc.citation.publicationname | APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | - |
dc.identifier.doi | 10.1007/s12010-014-1294-3 | - |
dc.contributor.localauthor | Chang, Yong-Keun | - |
dc.contributor.nonIdAuthor | Park, Da Yeon | - |
dc.contributor.nonIdAuthor | Chi, Won-Jae | - |
dc.contributor.nonIdAuthor | Park, Jae-Seon | - |
dc.contributor.nonIdAuthor | Hong, Soon-Kwang | - |
dc.description.isOpenAccess | N | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Pseudoalteromonas hodoensis | - |
dc.subject.keywordAuthor | beta-Agarase | - |
dc.subject.keywordAuthor | Neoagarotetraose | - |
dc.subject.keywordAuthor | Neoagarobiose | - |
dc.subject.keywordAuthor | Neoagarohexaose | - |
dc.subject.keywordAuthor | AgaH71 | - |
dc.subject.keywordPlus | MARINE BACTERIUM | - |
dc.subject.keywordPlus | ALPHA-AGARASE | - |
dc.subject.keywordPlus | EXTRACELLULAR AGARASE | - |
dc.subject.keywordPlus | ENZYMATIC-PROPERTIES | - |
dc.subject.keywordPlus | MOLECULAR-CLONING | - |
dc.subject.keywordPlus | STRAIN GJ1B | - |
dc.subject.keywordPlus | PURIFICATION | - |
dc.subject.keywordPlus | OLIGOSACCHARIDES | - |
dc.subject.keywordPlus | IDENTIFICATION | - |
dc.subject.keywordPlus | NEOAGAROBIOSE | - |
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