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http://dx.doi.org/10.4014/jmb.1710.10011

Biochemical Characterization of a Novel GH86 β-Agarase Producing Neoagarohexaose from Gayadomonas joobiniege G7  

Lee, Yeong Rim (Department of Bioscience and Bioinformatics, Myongji University)
Jung, Subin (Department of Bioscience and Bioinformatics, Myongji University)
Chi, Won-Jae (Biological and Genetic Resource Assessment Division, National Institute of Biological Resource)
Bae, Chang-Hwan (Biological and Genetic Resource Assessment Division, National Institute of Biological Resource)
Jeong, Byeong-Chul (Department of Bioscience and Bioinformatics, Myongji University)
Hong, Soon-Kwang (Department of Bioscience and Bioinformatics, Myongji University)
Lee, Chang-Ro (Department of Bioscience and Bioinformatics, Myongji University)
Publication Information
Journal of Microbiology and Biotechnology / v.28, no.2, 2018 , pp. 284-292 More about this Journal
Abstract
A novel ${\beta}$-agarase, AgaJ5, was identified from an agar-degrading marine bacterium, Gayadomonas joobiniege G7. It belongs to the glycoside hydrolase family 86 and is composed of 805 amino acids with a 30-amino-acid signal peptide. Zymogram analysis showed that purified AgaJ5 has agarase activity. The optimum temperature and pH for AgaJ5 activity were determined to be $30^{\circ}C$ and 4.5, respectively. AgaJ5 was an acidic ${\beta}$-agarase that had strong activity at a narrow pH range of 4.5-5.5, and was a cold-adapted enzyme, retaining 40% of enzymatic activity at $10^{\circ}C$. AgaJ5 required monovalent ions such as $Na^+$ and $K^+$ for its maximum activity, but its activity was severely inhibited by several metal ions. The $K_m$ and $V_{max}$ of AgaJ5 for agarose were 8.9 mg/ml and 188.6 U/mg, respectively. Notably, thin-layer chromatography, mass spectrometry, and agarose-liquefication analyses revealed that AgaJ5 was an endo-type ${\beta}$-agarase producing neoagarohexaose as the final main product of agarose hydrolysis. Therefore, these results suggest that AgaJ5 from G. joobiniege G7 is a novel endo-type neoagarohexaose-producing ${\beta}$-agarase having specific biochemical features that may be useful for industrial applications.
Keywords
Agar; ${\beta}$-agarase; GH86; Gayadomonas joobiniege;
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