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Characterization of a Novel Neoagarobiose-Producing GH42 β-Agarase, AgaJ10, from Gayadomonas joobiniege G7

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Abstract

Gayadomonas joobiniege G7 is an agar-degrading bacterium, which produces various agarases that have been biochemically characterized recently. In this study, we biochemically characterized a new β-agarase AgaJ10 belonging to the glycoside hydrolase (GH) 42 family from G. joobiniege G7. AgaJ10 is composed of 762 amino acids (89 kDa) and has the highest similarity (63% identity) to a putative β-agarase from the agar-degrading bacterium Catenovulum sp. DS-2, which was obtained from the intestines of a Haliotis diversicolor. The optimal pH and temperature for AgaJ10 activity were determined to be 5.0 and 30 °C, respectively. AgaJ10 exhibited a cold tolerance, retaining more than 40% of its enzymatic activity at 5 °C. The Km and Vmax of AgaJ10 for agarose were 61.5 mg/mL and 294.1 U/mg, respectively. Notably, the activity of AgaJ10 was significantly enhanced by Mn2+ but was strongly inhibited by some metal ions, including Fe2+, Ni2+, and Cu2+. Agarose-liquefaction, mass spectrometry, and thin-layer chromatography analyses showed that AgaJ10 is an exo-type β-agarase that hydrolyzes agarose only into neoagarobiose. Therefore, this study is the first report of a GH42 β-agarase that catalyzes a neoagarobiose-producing exo-type reaction.

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Funding

This work was supported by the 2016 Research Fund of Myongji University.

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  1. Department of Biological Sciences, Myongji University, Yongin, Gyeonggido, 449-728, Republic of Korea

    Umji Choi, Subin Jung, Soon-Kwang Hong & Chang-Ro Lee

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  1. Umji Choi
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Correspondence to Chang-Ro Lee.

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Choi, U., Jung, S., Hong, SK. et al. Characterization of a Novel Neoagarobiose-Producing GH42 β-Agarase, AgaJ10, from Gayadomonas joobiniege G7. Appl Biochem Biotechnol 189, 1–12 (2019). https://doi.org/10.1007/s12010-019-02992-5

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