• Korean Journal of Microbiology
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• v.53 no.1
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• pp.29-38
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• 2017

In this study, we investigated the phylogenetic diversity of the bacterial community and isolation of extremely halophilic bacteria using culturomics in a gray solar saltern. The number of bacterial living cells, enumerated in a gray solar saltern by direct fluorescence microscopy was three to four orders of magnitude greater than those enumerated by plate counts, suggesting the distribution of 'viable but non-culturable bacteria'. The biodiversity of bacterial communities in a gray solar saltern was investigated by pyrosequencing, 1,778 OTUs of bacteria were comprised of 18 phyla 46 classes 85 orders 140 families 243 genera with 6.16 diversity index. Archaea communities were composed of 3 phyla 6 classes 7 orders 7 families 38 genera with 4.95 diversity index from 643 OTUs. Totally 137 isolates were isolated by 59 different cultural methods based on culturomics considering culture media and conditions suitable for the growth of extremely halophilic bacteria. Phylogenetic analyses of extremely halophilic isolates based on 16S rRNA gene sequences, extremely halophilic isolates were composed of 4 phyla and 11 genera. Haloterrigena and Haloferax can be successfully isolated from culturomics. These culturomics were effective methods for collection of diversity of extremely halophilic bacteria.

• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.5
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• pp.856-862
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• 1994

In salt-preserved foods of every kinds, it was examined the growth condition of halophilic bacteria that induced a change of colour, taste, nutritive substance, a production condition of enzyme and a character of crude enzyme. Used bacteria is H. halobium a kind of extremely halophilic bacteria, and the required of optimum culture needed a quite long time of crude enzyme production is 168 hours. Optimum pH is about 7-7.5, so the traditional food of such neutrality pH as soybean paste and soy sauce particularly come into trouble because the growth can flourish in neutrality or alkaliescence, and the crude enzyme also appeared that best activation between pH 6 and pH 8. The optimum temperature is about 37$^{\circ}C$, the optimum temperature of enzyme is about 40 $^{\circ}C$ and the temperature stability is settled for 15 minutes and it is completely inactivated at 10 minutes. In the influence of each metal ion, Fe++ and Mn++ a stimulated the growth of H.halobium and the activation of enzyme, Cu++ and Zn++ were identified that made the growth and the activation of enzyme inhibit.

• Journal of Life Science
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• v.13 no.4
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• pp.505-510
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• 2003

The extremely halophilic archaebacteriurn Haloarcular sp. EH-1 was isolated from solar salts. Halophages found in Haloarcular sp. EH-1 were isolated from fermented anchovy sauce. Halophages were isolated from fermented anchovy sauce using Haloarcular sp. EH-1 as a host bacterium. The isolated halophage produced 0.5∼l.0 mm in diameter clear plaques. The halophage consists of an symmetrical head, measuring 68 nm in diameter, and a contractile tail, 100 nm long and base plates were observed. Total size of phage DNA genome obtained 20 Kbp and its sequence homology was 52.87% with H. Salinarium.

• Korean Journal of Microbiology
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• v.29 no.1
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• pp.56-62
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• 1991

Extremely halophilic bacteria isolated from salterns at Mado, Kyunggido, Korea, were identified and investigated on their salt requirements. The results have shown that six strains were identified to be belonged to the genus Halobacterium and three strains identified as the fenus Halococcus. Among them, the optimal NaCl concentration for growth of Halobacterium sp. EH10 was at 4.2M and no growth occurs below 2.0M NaCl. The strain, EH10, is nonmotile and showed acid production from glucose, fructose and maltose while H. salinarum is motile and does not produce acid from any carbohydrates. On the other hand, the strain EH10 does not utilize readily glucose while a number of sugars are readily utilized for growth with acid production by H. saccharovorum. Thus, the isolate, EH10, was classified into the genus Halobacterium and could be a novel species of the genus by its main morphological and physiological features including G+C content. The optimal temperature for growth of the isolate, EH10, was 50.deg.C. But this strain did not grow when NaCl was replaced with KCl.

• Journal of Life Science
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• v.12 no.5
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• pp.570-576
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• 2002

The extremely halophilic archaebacterium Haloarcular sp. EH-1 was isolated from solar salts. Amylae production from Halonrcular sp. EH-1 have been studied. The results obtained were as follows. The optimal medium composition for the production of amylase from Haloarcular sp. EH-1 were soluble starch 1.5%, yeast extract 1.0%, MgSO$_4$ 7h$_2$O 2.0%, KCI 0.1%, NaCl 25% (pH 7.5). The incubation temperature, aeration rate and agitation speed were 4$0^{\circ}C$, 100 $m\ell$ medium / 500 $m\ell$ shaking flask, and 110 rpm. The cell growth and enzymatic activity was highest at 9 days of incubation. So amylase production appeared to be a growth-related phenomenon.

• Microbiology and Biotechnology Letters
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• v.27 no.2
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• pp.129-135
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• 1999

EH-1 was highest at 9 days of incubation. This regrowth and enzymatic activity of Haloarcular sp. EH-1 was highest at 9 days of incubation. This amylase was purified by acetone fractionation, DEAG-Cellulose column chromatography, 1st Sephadex G-75 gel filtration, CM-Cellulose column chromatography and 2nd Sephadex G-75 gel filtration. The amylase was purified about 98.64 fold with a yield of 11.75%. The molecular weight of amylase was estimated to be about 43,000and 40,000 by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, suggesting that the enzyme was a monomer. Amylase had an optimal temperature of 4$0^{\circ}C$, and an optimum pH of 7.0, and the thermal stability was observed the above 50% at 10$0^{\circ}C$ after 1 hour, and the stable range of pH was 6.0 to 8.0. The enzymatic activity was increased in the presence of 10 mM 2-mercaptoethanol, slightly by 10 mM SnCl2.2H2O.FeCl2.4H2O.CuCl2.2H2O.HgCl2.6H2O and SDS. End products from soluble starch were glucose, maltose and maltotriose, and Km value for soluble starch was 2.5mg/ml.