To isolate aerobically and identify the diversity of halophilic bacteria in the soil around two ports, Daepopogu and Hwasun Port, on Jeju island, a total 46 halophilic bacteria strains were isolated and phylogenetically analysed. The isolated strains were divided into 3 phyla, 8 families, 16 genera and 23 species. The main taxa was the Bacilli class, which included 50.0% of the strains with 3 families, 10 genera and 15 species of Bacillaceae, Exiguobacterium_f and Planococcaceae. The second taxa was the Gammaproteobacteria class, which included 45.7% of the strains with 4 families, 5 genera and 7 species of Aeromonadaceae, Halomonadaceae, Marinobacteraceae and Vibrionaceae. The isolated strains were tested for hydrolytic enzymes, amylase, lipase and protease activity, and 31 strains showed activity of at least one enzyme. Furthermore, auxin activity was determined in 7 strains. This study showed that the isolated strains have possible applications in the food and agricultural industries and have importance as a genetic resource in Korea.
본 연구는 국내외 해양유래 발효산물 시료로부터 분리한 호염성 미생물들의 다양성 및 특성에 관하여 조사하였다. 호염성 미생물의 순수 분리를 위하여 marine agar 배지를 사용하였으며 25, 37, 50oC에서 호기적으로 배양하였다. 순수 분리 후, 86균주를 분리하였으며 16S rRNA 염기서열 분석 결과를 바탕으로 계통학적 분석을 실시한 결과, 3문, 7과, 9속, 24종으로 구성되어 있는 것을 확인하였다. 특히 Firmicutes문 Bacilli강은 84.9%의 분포를 나타내었으며 4과, 6속, 19종으로 Bacillaceae, Planococcaceae, Staphylococcaceae와 Enterococcaceae로 분포하는 것을 확인하였다. 그리고 분리한 균주들이 amylase, lipase, cellulase, protease 같은 산업적으로 유용한 효소를 생산하는지 확인하기 위하여 효소 활성 평가를 실시하였으며, 55 균주가 최소 한 종류 이상의 효소 활성을 가지고 있는 것을 확인하였다. 또한 옥신 생산능을 가지는 균주도 2 균주가 확인되었으며 이는 본 연구를 통하여 분리한 미생물들의 산업적 활용 가능성을 나타내었다. 그러므로 이번 연구는 국내 유전자원 확보 및 시료의 호염성 미생물의 다양성과 특성에 관한 과학적 지식 확장에 도움이 될 것으로 생각된다.
Screening of Bacillus subtilis strains capable of producing extracellular hydrolases, including lipolytic enzyme was carried out. Within the scope of this study, total 61 strains were isolated from various domestic specific environmental samples such as seawater, soil, Jeju ranch, hot spring, salt farm, and fermented food and identified as Bacillus subtilis based on 16S rRNA sequences. Among the isolates, 4 strains had extracellular lipase activity. The growth profile of the strains revealed that 4 strains showed well growth at 50℃, 3 strains at 55℃, and pH 7.0. All strains could tolerate salinity up to 5%(w/v), 3 strains up to 10%(w/v), and only one strain showed growth at 15%(w/v). Additionally, amylolytic and proteolytic activities were detected in these strains. The highest lipolytic, amylolytic, and proteolytic activity was detected in Bacillus subtilis YS-YR 5A. These results demonstrate the potential application of extracellular hydrolase-producing Bacillus subtilis strains, especially strain YS-YR 5A, as fermentation starter to enhance the functionality and multiplication of functional natural products in the food/medicine/cosmetics/bio industries.