서해안 해수로부터 분리한 한천분해 해양미생물 Pseudoalteromonas sp. H9의 동정 및 특성 연구
Isolation and Characterization of an Agar-hydrolyzing Marine Bacterium, Pseudoalteromonas sp. H9, from the Coastal Seawater of the West Sea, South Korea
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초록
대한민국 대천 해수로부터 agarase를 생산하는 균주 H9 을 분리하였다. 본 균주는 16S rRNA 염기 염기서열 분석결과로부터 Pseudoalteromonas espejiana NCIMB2127T (98.98%), Pseudoalteromonas carrageenovora ATCC12662T (98.78%), Pseudoalteromonas atlantica IAM12927T (98.64%), Pseudoalteromonas issachenkonii KMM3549T (98.63%) 등과 높은 상동성을 보였다. 균주 H9은 genomic DNA 내 G+C 농도가 41.56%이고 주요 퀴논으로 quinone- 8을 포함하고 있다. 균주 H9의 주요 지방산으로 C16:1ω7c (34.3%), C16:0 (23.72%), C18:1ω7c (13.64%) 등이 포함되었다. 이러한 유전적, 생리적 특성에 따라 균주 H9은 Pseudoalteromonas 속의 균으로 분류하여 Pseudoalteromonas sp. H9으로 명명하였다. 균주 H9이 세포외부로 분비하는 총 agarase는 40−45℃와 pH 7.0−8.0의 조건에서 높은 효소 활성을 갖으며, agarose를 분해하여 (neo)agarotetraose와(neo)agarohexaose를 생산하였다. 균주 H9은 한천분해를 위해 유용하게 사용될 수 있으며, 다양한 생리활성을 갖는 (neo)agarooligosaccharide는 기능성 식품, 화장품 등의 산업에 유용하게 사용될 수 있을 것으로 기대된다.
Abstract
An agarolytic marine bacterium (H9) was isolated from the coastal seawater of the West Sea, South Korea. The isolate, H9, was gram-negative and rod-shaped with a smooth surface and polar flagellum. Cells grew at 20−30℃, between pH 5.0 and 9.0, and in ASW-YP (Artificial Sea Water-Yeast extract, Peptone) media containing 1−5% (w/v) NaCl. The G+C content was 41.56 mol%. The predominant isoprenoid quinone in strain H9 was ubiquinone-8. The major fatty acids (>10%) were C16:1ω7c (34.3%), C16:0 (23.72%), and C18:1ω7c (13.64%). Based on 16S rRNA gene sequencing, and biochemical and chemotaxonomic characterization, the strain was designated as Pseudoalteromonas sp. H9 (=KCTC23887). In liquid culture supplemented with 0.2% agar, the cell density and agarase activity reached a maximum level of OD = 4.32 (48 h) and OD = 3.87 (24 h), respectively. The optimum pH and temperature for the extracellular crude agarases of H9 were 7.0 and 40℃, respectively. Thin-layer chromatography analysis of the agarase hydrolysis products revealed that the crude agarases hydrolyze agarose into neoagarotetraose and neoagarohexaose. Therefore, the new agar-degrading strain, H9, can be applicable for the production of valuable neoagarooligosaccharides and for the complete degradation of agar in bio-industries.
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