DC Field | Value | Language |
---|---|---|
dc.contributor.author | Chi, Won-Jae | ko |
dc.contributor.author | Park, Da Yeon | ko |
dc.contributor.author | Seo, Young Bin | ko |
dc.contributor.author | Chang, Yong Keun | ko |
dc.contributor.author | Lee, Soon-Youl | ko |
dc.contributor.author | Hong, Soon-Kwang | ko |
dc.date.accessioned | 2014-08-29T04:25:54Z | - |
dc.date.available | 2014-08-29T04:25:54Z | - |
dc.date.created | 2014-06-09 | - |
dc.date.created | 2014-06-09 | - |
dc.date.issued | 2014-05 | - |
dc.identifier.citation | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.98, no.10, pp.4545 - 4555 | - |
dc.identifier.issn | 0175-7598 | - |
dc.identifier.uri | http://hdl.handle.net/10203/189072 | - |
dc.description.abstract | Alteromonas sp. GNUM-1 is known to degrade agar, the main cell wall component of red macroalgae, for their growth. A putative agarase gene (agaG1) was identified from the mini-library of GNUM-1, when extracellular agarase activity was detected in a bacterial transformant. The nucleotide sequence revealed that AgaG1 had significant homology to GH16 agarases. agaG1 encodes a primary translation product (34.7 kDa) of 301 amino acids, including a 19-amino-acid signal peptide. For intracellular expression, a gene fragment encoding only the mature form (282 amino acids) was cloned into pGEX-5X-1 in Escherichia coli, where AgaG1 was expressed as a fusion protein with GST attached to its N-terminal (GST-AgaG1). GST-AgaG1 purified on a glutathione sepharose column had an apparent molecular weight of 59 kDa on SDS-PAGE, and this weight matched with the estimated molecular weight (58.7 kDa). The agarase activity of the purified protein was confirmed by the zymogram assay. GST-AgaG1 could hydrolyze the artificial chromogenic substrate, p-nitrophenyl-beta-d-galactopyranoside but not p-nitrophenyl-alpha-d-galactopyranoside. The optimum pH and temperature for GST-AgaG1 activity were identified as 7.0 and 40 A degrees C, respectively. GST-AgaG1 was stable up to 40 A degrees C (100 %), and it retained more than 70 % of its initial activity at 45 A degrees C after heat treatment for 30 min. The K (m) and V (max) for agarose were 3.74 mg/ml and 23.8 U/mg, respectively. GST-AgaG1 did not require metal ions for its activity. Thin layer chromatography analysis, mass spectrometry, and C-13-nuclear magnetic resonance spectrometry of the GST-AgaG1 hydrolysis products revealed that GST-AgaG1 is an endo-type beta-agarase that hydrolyzes agarose and neoagarotetraose into neoagarobiose. | - |
dc.language | English | - |
dc.publisher | SPRINGER | - |
dc.subject | ALPHA-AGARASE | - |
dc.subject | STRUCTURAL-ANALYSIS | - |
dc.subject | AGAROSE OLIGOMERS | - |
dc.subject | GENUS ALTEROMONAS | - |
dc.subject | MARINE BACTERIUM | - |
dc.subject | STRAIN GJ1B | - |
dc.subject | PURIFICATION | - |
dc.subject | NEOAGAROBIOSE | - |
dc.subject | GALACTANS | - |
dc.subject | PRODUCTS | - |
dc.title | Cloning, expression, and biochemical characterization of a novel GH16 beta-agarase AgaG1 from Alteromonas sp GNUM-1 | - |
dc.type | Article | - |
dc.identifier.wosid | 000335460700023 | - |
dc.identifier.scopusid | 2-s2.0-84900835642 | - |
dc.type.rims | ART | - |
dc.citation.volume | 98 | - |
dc.citation.issue | 10 | - |
dc.citation.beginningpage | 4545 | - |
dc.citation.endingpage | 4555 | - |
dc.citation.publicationname | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | - |
dc.identifier.doi | 10.1007/s00253-014-5510-4 | - |
dc.contributor.localauthor | Chang, Yong Keun | - |
dc.contributor.nonIdAuthor | Chi, Won-Jae | - |
dc.contributor.nonIdAuthor | Park, Da Yeon | - |
dc.contributor.nonIdAuthor | Lee, Soon-Youl | - |
dc.contributor.nonIdAuthor | Hong, Soon-Kwang | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Alteromonas sp GNUM-1 | - |
dc.subject.keywordAuthor | beta-Agarase | - |
dc.subject.keywordAuthor | Neoagarobiose | - |
dc.subject.keywordAuthor | AgaG1 | - |
dc.subject.keywordPlus | ALPHA-AGARASE | - |
dc.subject.keywordPlus | STRUCTURAL-ANALYSIS | - |
dc.subject.keywordPlus | AGAROSE OLIGOMERS | - |
dc.subject.keywordPlus | GENUS ALTEROMONAS | - |
dc.subject.keywordPlus | MARINE BACTERIUM | - |
dc.subject.keywordPlus | STRAIN GJ1B | - |
dc.subject.keywordPlus | PURIFICATION | - |
dc.subject.keywordPlus | NEOAGAROBIOSE | - |
dc.subject.keywordPlus | GALACTANS | - |
dc.subject.keywordPlus | PRODUCTS | - |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.