• Applied Biological Chemistry
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• v.34 no.4
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• pp.307-311
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• 1991

A moderate halophile, ES 10 which produces a high level of alkaline protease was isolated from the salted anchovies and indentified as a strain of Halomonas sp. The optimum growth of the Halomonas sp. was revealed in the presence of 2 M NaCl and its growth rate in the Temporary Synthetic Medium was increased by adding DL-alanine, but inhibited by adding L-proline. The concentration of $Na^+$, $K^+$ and $Mg^{2+}$ in the cell mass of the Halomonas sp. ES 10 was 5-, 25- and 35-fold higher by dry weight basis, respectively than those of B. subtilis or E. coli. Norberg and Hofsten medium with 1 M NaCl was selected as the best medium for producing high level of alkaline protease. The optimum temperature for the growth and protease production was equally $20^{\circ}C$.

• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.51-55
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• 1997

To utilize the alkaline protease produced by Halomonas sp. ES-10 as an enzyme detergent, the crude enzyme was obtained by methanol precipitation and lyophilization. And it was processed to coated enzyme.The best mixing ratio of components such as coated enzyme, builders, actives, fillers and adjuvants on detergency was examined, and temperature and pH influencing detergency were also tested. Detergency test 0.15% detergent solution was carried out on EMPA test cloth #116 with shaking(90 rpm) for 10 min after 30 min of pretreatment. The detergent which contained coated-enzyme 1%, Zeolite 4A 20%, Tween 80 1. 5%, sodium borate 30%, sodium meta silicate 7.5% and water 40% showed about 90% of washing efficiency at 40$\circ $C and pH 10.0.

• Journal of Applied Biological Chemistry
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• v.48 no.2
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• pp.75-78
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• 2005

Halotolerant protease produced by Halomonas marisflava KCCM 10457 was partially purified through ammonium sulfate precipitation and Sephacryl S-200HR gel permeation chromatography. Optimal pH and temperature of protease were 11.0 and $45^{\circ}C$, respectively. Enzyme activity was inhibited by $Cu^{2+}$, $Hg^{2+}$, $Fe^{2+}$, and $Fe^{3+}$, and selectively inhibited by p-chloromercuribenzoic acid (PCMB), suggesting this enzyme is cysteine protease. The enzyme is halotolerant, because it retained 77% of original activity in presence of 3.33 M NaCl. The protease showed broad substrate specificity to various natural proteins; BSA, casein, egg albumin, gelatin, and hemoglobin.

• Proceedings of the Korean Institute of Building Construction Conference
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• 2020.11a
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• pp.159-160
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• 2020

This study examined the potentials for developing a biological coating material with high chloride resistance. The bacteria strains isolated were Halomonas alkaliphile, Halomonas venusta, and Sulfidobacter mediterraneus. Test results revealed that the developed approach is very promising in reducing the chloride ion diffusion coefficient of concrete.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.250-258
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• 2021

Among various species of marine bacteria, those belonging to the genus Halomonas have several promising applications and have been studied well. However, not much information has been available on their antibiotic resistance. In our efforts to learn about the antibiotic resistance of strain Halomonas socia CKY01, which showed production of various hydrolases and growth promotion by osmolytes in previous study, we found that it exhibited resistance to multiple antibiotics including kanamycin, ampicillin, oxacillin, carbenicillin, gentamicin, apramycin, tetracycline, and spectinomycin. However, the H. socia CKY01 resistance pattern to kanamycin, gentamicin, apramycin, tetracycline, and spectinomycin differed in the presence of 10% NaCl and 1% NaCl in the culture medium. To determine the mechanism underlying this NaCl concentration-dependent antibiotic resistance, we compared four aminoglycoside resistance genes under different salt conditions while also performing time-dependent reverse transcription PCR. We found that the aph2 gene encoding aminoglycoside phosphotransferase showed increased expression under the 10% rather than 1% NaCl conditions. When these genes were overexpressed in an Escherichia coli strain, pETDuet-1::aph2 showed a smaller inhibition zone in the presence of kanamycin, gentamicin, and apramycin than the respective control, suggesting aph2 was involved in aminoglycoside resistance. Our results demonstrated a more direct link between NaCl and aminoglycoside resistance exhibited by the H. socia CKY01 strain.

• Journal of Microbiology
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• v.42 no.3
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• pp.174-180
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• 2004

A halophilic bacterium was isolated from fermented seafood. The 16S rDNA sequence identity between the isolate and Halomonas subglaciescola AJ306801 was above 95％. The isolate that did not grow in the condition without NaCl or in the condition with other sodium (Na$\^$+/) or chloride ions (Cl$\^$-/) instead of NaCl was named H. subglaciescola DH-l. Two mutants capable of growing without NaCl were obtained by random mutagenesis, of which their total soluble protein profiles were compared with those of the wild type by two-dimensional electrophoresis. The external compatible solutes (betaine and choline) and cell extract of the wild type did not function as osmoprotectants, and these parameters within the mutants did not enhance their growth in the saline environment. In the proton translocation test, rapid acidification of the reactant was not detected for the wild type, but it was detected for the mutant in the condition without NaCl. From these results, we derived the hypothesis that NaCl may be absolutely required for the energy metabolism of H. subglaciescola DH-l but not for its osmoregulation, and the mutants may have another modified proton translocation system that is independent of NaCl, except for those mutants with an NaCl-dependent system.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.118-124
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• 2005

In this research, we examined the effect of NaCl on the growth, energy metabolism, and proton motive force of Halomonas salina, and the effect of compatible solutes on the bacterium growing in the high salinity environment. H. salina was isolated from seawater and identified by 16srDNA sequencing. The growth of H. salina was not enhanced by the addition of external compatible solutes (choline and betaine) in the high salinity environment. The resting cells of H. salina absorbed more glucose in the presence of 2.0 M NaCl than in its absence. H. salina did not grow in the medium with either KCl, RbCl, CsCl, $Na_2SO_4$, or $NaNO_3$, in place of NaCl. The optimal concentration of NaCl for the growth of H. salina ranged from 1.4 M to 2.5 M, and the growth yield was decreased in the presence of NaCl below 1.4M and above 2.5M. The activity of isocitrate dehydrogenase, pyruvate dehydrogenase, and malate dehydrogenase of H. salina was not inhibited by NaCl in in vitro test. The proton translocation of H. salina was detected in the presence of NaCl only. These results indicate that NaCl is absolutely required for the normal growth and energy metabolism of H. salina, but the bacterial growth is not enhanced by the compatible solutes added to the growth medium.

• Applied Biological Chemistry
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• v.35 no.4
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• pp.237-241
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• 1992

The protease from Halomonas sp. ES 10 was purified by methanol precipitation, gel filtration on Sephadex G-150 and G-200, and ion exchange chromatography on DEAE-Sephadex A-50. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis. The specific activity of purified enzyme was 1,014 units/mg protein, and the yield of the total activity from the culture filtrate was 7%. The optimal temperature and pH for the enzyme activity were $35^{\circ}C$, and pH 11.0, respectively. And the enzyme was stable in the range of $pH\;7.5{\sim}11.0$. The residual activity of the enzyme was 70%, when the enzyme was incubated at $50^{\circ}C$ for 40 min. The Km value of the enzyme was 7.4 mg/ml to milk casein. $Li^+$, $Ca^{2+}$, SDS and Tween 80 were appeared to activators, whereas $Hg^{2+}$ and EDTA to inhibitors. The addition of DFP and PMSF showed the relative enzyme activities of 63% and 107%, respectively, suggesting that the enzyme may not belong to serine type protease. When the alkaline protease was treated with 0.5 M and 1 M NaCl, the relative enzyme activities were 95% and 65%, respectively. This enzyme showed 20% and 15% higher enzyme activity than that of Aspergillus oryzae (Sigma Chemical Company product, P4755) in the presence of 0.5 M and 1 M NaCl.

• Proceedings of the Korean Institute of Building Construction Conference
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• 2022.04a
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• pp.49-50
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• 2022

This study examined the potential of halophilic bacteria in reducing the chloride ion concentration within the cement mortars exposed to chloride attack. As a result of the experiment, the compressive strength of mortar with Halomonas venusta showed an equivalent performance to that of counterpart mortars without bacteria. Also, the chloride ion concentration measured in mortars including Halomonas Venusta was 71% lower than that of the counterpart mortars without bacteria.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.298-303
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• 2007

A halophilic bacterium, H. subglaciescola DH-1, grew at 2.0 M salinity, but did not grow at 0.8 M salinity when cultivated at higher temperature ($40^{\circ}C$) than optimum ($30^{\circ}C$). When the cell extract of strain DH-1 was heated at $50^{\circ}C$ for 60 min in the absence of NaCl, isocitrate dehydrogenase and malate dehydrogenase lost their activities, but when it was heated in the presence of 2.0 M NaCl, the activity was maintained. Meanwhile, the cell extract of E. coli did not catalyze the reduction of $NAD^+$ to NADH coupled with the oxidation of isocitrate and malate at higher salinities than 1.0 M. The pH range for DH-1 was 7 to 10, and that for E. coli was 5 to 9. DH-1 was not grown in conditions with sodium salts other than NaCl.