title: Molecular cloning, characterization and enzymatic properties of a novel βeta-agarase from a marine isolate Psudoalteromonas SP. AG52.
abstract: An agar-degrading Pseudoalteromonas sp. AG52 bacterial strain was identified from the red seaweed Gelidium amansii collected from Jeju Island, Korea. A β-agarase gene which has 96.8% nucleotide identity to Aeromonas β-agarase was cloned from this strain, and was designated as agaA. The coding region is 870 bp, encoding 290 amino acids and possesses characteristic features of the glycoside hydrolase family (GHF)-16. The predicted molecular mass of the mature protein was 32 kDa. The recombinant β-agarase (rAgaA) was overexpressed in Escherichia coli and purified as a fusion protein. The optimal temperature and pH for activity were 55 °C and 5.5, respectively. The enzyme had a specific activity of 105.1 and 79.5 unit/mg toward agar and agarose, respectively. The pattern of agar hydrolysis demonstrated that the enzyme is an endo-type β-agarase, producing neoagarohexaose and neoagarotetraose as the final main products. Since, Pseudoalteromonas sp. AG52 encodes an agaA gene, which has greater identity to Aeromonas β-agarase, the enzyme could be considered as novel, with its unique bio chemical characteristics. Altogether, the purified rAgaA has potential for use in industrial applications such as development of cosmetics and pharmaceuticals.
authors.affiliation: Department of Marine Life Science, Jeju National University 66 Jejudaehakno , Ara-Dong, Jeju, 690-756 , Republic of Korea ; Korea Ocean Research & Development Institute , Ansan, 426-744 , Republic of Korea.
authors.forename: Chulhong
authors.initials: C
authors.lastname: Oh
authors.forename: Chamilani
authors.initials: C
authors.lastname: Nikapitiya
authors.forename: Youngdeuk
authors.initials: Y
authors.lastname: Lee
authors.forename: Ilson
authors.initials: I
authors.lastname: Whang
authors.forename: Do-Hyung
authors.initials: DH
authors.lastname: Kang
authors.forename: Soo-Jin
authors.initials: SJ
authors.lastname: Heo
authors.forename: Young-Ung
authors.initials: YU
authors.lastname: Choi
authors.forename: Jehee
authors.initials: J
authors.lastname: Lee
keywords: Aeromonas sp.; Agar; GHF-16; Neoagaro-oligosaccharides; Pseudoalteromonas sp.; β-agarase
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