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Microbiology and Biotechnology Letters


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Fermentation Microbiology (FM)  |  Fermentation Technology

Microbiol. Biotechnol. Lett. 2021; 49(3): 329-336

Received: May 12, 2021; Revised: July 5, 2021; Accepted: July 7, 2021

해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특

Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3

Da Som Kim1, Jong-Hee Kim2 and Won-Jae Chi1*

1Microorganism Resources Division, National Institute of Biological Resource, Incheon 22689, Republic of Korea 2Department of Food and Nutrition, Seoil University, Seoul 02192, Republic of Korea

Correspondence to :
Won-Jae Chi,

Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 °C. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

Keywords: Cellulophga sp. J9-3, neoagarotetraose, neoagarohexaose, agarose, agar

Graphical Abstract

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