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.

The diversity and characterization of microorganisms isolated from the soil around several ranches in Korea were confirmed in this study. To isolate halophilic microorganisms, the marine agar medium was basically used and cultivated at 37℃ for several days. After single colony isolation, a total of 116 pure colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rDNA sequencing, indicating that isolated strains were divided into 4 phyla, 23 families, 30 genera and 51 species. To confirm whether isolated strain can be a candidate for the fermentation of diverse food ingredients, amylase, lipase, and protease enzyme assays were performed individually, showing that 92 strains possessed at least one enzyme activity. Especially 4 strains, identified to Jonesia quinghaiensis (isolate name: JSF 19-2), Halomonas alkaliantarctica (isolate name: JSF 21), Bacillus velezensis (isolate name: NWFY-36), and Staphylococcus capitis subsp. urealyticus (isolate name: MSY-5), showed all enzyme activity tested. Moreover, 17 strains showed the ability for auxin production. This result indicated that isolated strains have shown the possibility of the application for the food and feed industries. Therefore, this study has contributed to securing domestic biological resources and the improvement of hydrolytic enzyme activity by using isolated strains.

This research confirmed the diversity and characterization of halophilic microorganisms isolated from the various solar salterns, collected on the inside and outside of the country. To isolate strains, the marine agar medium was basically used and cultivated at 37oC for several days aerobically. After single colony isolation, a totally of 230 pure colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rDNA sequencing, indicating that isolated strains were divided into 4 phyla, 12 families, 27 genera and 64 species. Firmicutes phylum, the main phyletic group, comprised 89.6% with 3 families, 17 genera and 52 species of Bacillaceae, Staphylococcaceae and Carnobacteriaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 177 strains possessed at least one enzyme activity. Especially 17 strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed to securing domestic genetic resources and the expansion of scientific knowledge of the halophilic microorganisms community in solar salterns.

A novel halophilic archaeon designated strain CBA1114T was isolated from solar salt in the Republic of Korea. Strain CBA1114T, which is a coccoid and stained Gram-negative, grew in the presence of 15-30% (w/v) NaCl (optimum, 20%) and at 20-50°C (optimum, 40°C) and pH 7.0-9.0 (optimum, pH 8.0). Strain CBA1114T required Mg2+ for growth. Strain CBA1114T had three 16S rRNA genes, rrnA, rrnB and rrnC; similarities between the sequences were 99.7 and 99.9%. The 16S rRNA gene sequence of strain CBA1114T showed a 91.7% similarity to that of Haloterrigena thermotolerans PR5T. In multilocus sequence analysis (MLSA), five housekeeping genes, atpB, EF-2, radA, rpoB’ and secY, were found to be closely related to those of the members of the genera Halorientalis (89.7% similarity of the atpB gene sequence), Halomicroarcula (91.9 %, EF-2), Haloterrigena (85.4 %, radA), Natronoarchaeum (89.2 %, rpoB’) and Natrinema (75.7 %, secY). A phylogenetic tree generated from the results of 16S rRNA gene and MLSA of the five housekeeping genes showed that strain CBA1114T was closely related to the species of the genus Halorientalis in the family Halobacteriaceae. The major polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and unidentified lipids. According to the results of phylogenetic, phenotypic and chemotaxonomic analyses, we designate strain CBA1114T as Halostella salina gen. nov., sp. nov., which represents a novel species of a novel genus within the family Halobacteriaceae.

This research is on the halophilic bacteria inhibiting in solar salterns. It was conducted by measuring the composition of cultivating ground in each step of salt production and targeting solar salterns during 2012 and 2013 in Jeungdo, Shinangun, JeollaNam-do, Korea. In each step of salt production, the succession of the group has been studied using pyrosequencing method and the new halophile variety has been separately identified in solar salterns. The results are as follows; The decomposition ability of protein, lipid, cellulose, and starch, has been experimented with separately identifying 33 bacteria that appeared as dominants in solar salterns. Overall, the bacteria with decomposition capability of all 4 components were Micrococcus luteus, Nocardioides daejeonensis in Actinobacteria group, Halobacillus trueperi whb45 in Firmicutes group, and Aurantimonas coralicida, Lutibacterium anuloederans, Nesiotobacter exalbescens in Alphaproteobacteria group. Analysis in the diversity of bacteria community of diluted solution with concentration of 3%, 6%, 11%, 20%, and 30% has been made using pyrosequencing method. In the Order level, SAR 11 dominated in 3% salt solution, Rhodobacterales group (18.9%) in 6% solution, Micrococcales group showed the highest ratio in 11% and 20% solutions, and Rhodobacterales group was dominant in the solution of 30%. In Phylum level, Proteobacteria group showed dominancy in 3% and 6% salt solutions, Actinobacteria group was dominant in 11% and 20% solutions, and Proteobacteria group was dominant in 30% solution. Among the isolated strains, seven novel strains (Kmb21, Kmb26, Kmb27, Kmb32, Kmb42, Kmb64, and Kmb80) were donate registered to NCBI and KCTC, JCM, each as new variety candidate.

피부는 외부 유해물질로부터 내부기관을 보호하는 장벽기능을 하는 대표적인 기관으로 자외선 (ultraviolet radiation, UV), 중금속, 각종 산화 물질들과 같은 외부의 위협에 항상 노출되어 있어 손상을 받 기 쉽다. 특히 자외선 B (UVB)는 진피의 상부까지 도달하여 화상이나 홍반과 같은 염증반응을 일으키며 멜라 닌 생성을 촉진시켜 색소 침착을 유발한다. 지나친 자외선 B의 피부세포로의 유입은 각질세포 및 진피 섬유아 세포의 DNA손상을 야기하고, 세포외기질의 합성을 방해하여 피부탄력감소, 주름생성, 진피 결합조직의 손상 과 함께 피부장벽을 붕괴시켜 노화를 일으키며, 장기간 노출 시 심각한 피부 병변으로 이어져 피부세포 사멸 및 종양의 발생으로까지 이어진다. Haloarcula vallismortis는 사해로부터 분리 동정한 미생물로 호염성 고생 물의 생장적 특징은 아직 자세히 연구된 바는 없다. 대게 10% 이상의 염도에서 자라는데 실제 생장염도는 평균 20 ∼ 25% 염도에서 자란다. 염도가 높은 호수나 염전에서 주로 발견되기 때문에 강한 햇빛에 대한 방 어기작이 존재한다. 그중 하나로 다른 ATP를 생성하기 위한 bacteriorhodopsin외에 halorhodopsin이라는 색소를 이용, 광자(photon)를 흡수하고 염화이온 채널을 개방시켜 생기는 전위차를 이용해 추가로 에너지를 생산한다. 또 carotenoid 색소로 인해 농도가 높을시 분홍색 또는 빨간색을 띄는 특징이 있는데 이것이 강한 자외선에 대한 방어기작을 할 것이라고 여겨진다. 본 연구는 호염성 고세균이 자외선을 에너지 소스로 이용하 는 특성을 이용하여 자외선으로부터 피부를 방어하는 방법을 개발하고자 이들의 피부각질세포에서 자외선에 의한 항염 효과 및 DNA 손상 방어 효과를 확인하였다. 그리고 이들을 천연 자외선 차단제의 소재로서 사용 할 수 있음을 확인하였다.