• 한국신재생에너지학회:학술대회논문집
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• 2009.06a
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• pp.782-786
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• 2009

Thiocapsa roseopersicina NCIB 8347은 purple sulfur bacteria이며 광합성종속영양 조건에서는 nitrogenase 효소계가 유도되어 질소를 고정하며, 수소를 발생한다. 또한 광합성독립영양 조건에서는 hydrogenase 효소계가 유도되어 3~4개 종류의 특성이 다른 hydrogenase가 membrane에 결합되어 있거나, cytoplasma에 존재하며, 이 중의 일부는 산소농도와 온도의 상승에도 비교적 안정하다. 본 연구에서는 T. roseopersicina NCIB 8347이 광합성종속영양 조건에서 수소를 생산할 수 있는 제반 배양조건을 최적화하고, nitrogenase와 일부 hydrogenase역가를 측정하여 purple non-sulfur bacteria, Rhodobacter sphaeroides KD131의 nitrogenase와 비교하여 수소생산을 최적화하였다. 할로겐램프를 8-9 $Klux/m^2$로 조사할 때와 배양온도 $26{\sim}30^{\circ}C$, 배양시간 72시간에서 균체 성장과 수소생산이 가장 높았다. T. roseopersicina NCIB 8347는 광합성 독립영양, 종속영양 조건에서 모두 성장 할 수 있었다.

• Transactions of the Korean hydrogen and new energy society
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• v.17 no.4
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• pp.371-378
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• 2006

Effect of $(NH_4)_2SO_4$ precipitation and heat-treatment on hydrogenase which was extracted from the cytoplasmic fraction of the phototrophic purple sulfur bacterium Thiocapsa roseopersicina NCIB 8347 was studied. Crude enzyme extract was prepared by centrifugation($28,000{\times}g$, $400,000{\times}g$) after sonication of cells grown under photosynthetic condition for 96 hrs. Various conditions of $(NH_4)_2SO_4$ precipitation and heat-treatment were examined and the effect of protein concentration was analyzed by SDS-electrophoresis between the treatments. Optimum conditions for $(NH_4)_2SO_4$ precipitation and heat-treatment for evolution hydrogenase activity were 40-60% saturation and $60^{\circ}C$ for 20 min, respectively, which exhibited the specific hydrogenase activity of 0.78 U/mg-protein. Specific hydrogenase activity was decreased to 31.6% when the heat-treatment at $60^{\circ}C$ increased from 20 min to 5 hrs.

• Applied Chemistry for Engineering
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• v.20 no.4
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• pp.449-452
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• 2009

The absence of Fe, Se, and Mo in a minimal medium prevented the production of hydrogen from the anaerobic culture of Escherichia coli MC4100. Fe, Se, and Mo are known to be cofactors of formate dehydrogenase ($FDH_{II}$) of both E. coli and Enterobacter aerogenes. Hence when these trace elements are absent in the minimal medium, hydrogen production through formate dehydrogenation would be inhibited not only in E. coli but also in E. aerogenes. Hydrogen production by E. aerogenes 413 was delayed when lacking these trace elements. Therefore, it is believed that hydrogen production of E. aerogenes is initiated not by the reoxidation of nicotinamide adenine dinucleotide (NADH) but by formate decarboxylation.

• KSBB Journal
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• v.20 no.6
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• pp.447-452
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• 2005

The hydrogen production using immobilized cellsl was conducted using fruit wastewaters at various culture conditions. Three kinds of fruit wastewaters, melon, watermelon and pear were used. Sodium alginate was used as immobilization material. Among them, concentration of reducing sugar which was one of the main components in fruit was the highest at watermelon wastewater, and also hydrogen production was the highest as 2319.2 mL/L in it. Although hydrogen production was not much changed according to sodium alginate concentration, its production was the most at 3%(w/v). As bead size as small, hydrogen production was higher. With inspection of interior, it confirmed that the cell grew well in bead. But the addition of amino acids using as agent for metabolite production had almost no affected on hydrogen productivity. The effective range of $FeSO_4$ addition on hydrogen production were up to 1.2 g/L, and above the concentration, it inhibited the productivity. Organic acids produced during watermelon fermentation were mainly lactic acid, butyric acid, abd acetic acid; and a little of propionic acid.

• Transactions of the Korean hydrogen and new energy society
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• v.19 no.2
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• pp.124-131
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• 2008

Crude cytoplasmic fraction of phototrophic purple sulfur bacterium, Thiocapsa roseopersicina NCIB 8347, were initially prepared and purified by sonication, ultracentrifugation, ammonium sulfate fractionation and heat-treatment and it has been previously reported. Using various applications of chromatography far the purification of membrane-bound and soluble hydrogenases from heat-treated enzyme fraction were studied at present report. When the heat-treated enzyme preparation was applied to the anion column chromatography using Q-sepharose, Fraction I and II, which were extracted with the KCl 0-0.5 M gradient, showed the specific evolution hydrogenase activity 3.86 and 2.27 U/mg-protein respectively. Specific hydrogenase activitys of Fraction I and II were further increased to 4.35 and 7.46 U/mg-protein for Fraction I and to 2.49 and 4.41 U/mg-protein fur Fraction II respectively, when hydrophobic interaction column, Phenyl superose, and anion exchange column, Mono-Q, were applied. Size exclusion chromatography using superdex 200 concentrated the hydrogenase Fraction I and II to 9.19 and 7.84 U/mg-protein respectively at the final step of purification.

• KSBB Journal
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• v.20 no.6
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• pp.413-417
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• 2005

The purple sulfur phototrophic bacterium, Thiocapsa roseopersicina NCIB 8347 has been studied on hydrogen production and cell growth under different culture conditions, such as light source, light intensity, and growth temperature. T. roseopersicina showed maximum cell growth of 1.38 and 1.42 g-DCW/L under 7.5-10 klux of halogen and fluorescent light, respectively, and produced maximum amount of hydrogen with values of 0.90 and 0.48 $mL-H_2/mg$-DCW under the irradiation of 10 klux of halogen and fluorescent light, respectively. The optimum growth temperature for hydrogen production was $26^{\circ}C$, and hydrogen production rate was lowered over $30^{\circ}C$. When T. roseopersicina was grown photoheterotrophically under irradiation of 8-9 klux of halogen lamp, the generation time was 4.2 hr. The strains started producing hydrgen from the middle of the logarithmic growth phase and continued until succinate concentration leveled out.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.446-450
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• 2004

We investigated the effective culture conditions of anaerobic bacteria, Enterobacter cloacae YJ-1 on hydrogen production. It was cultured with 60 mL of working volume at $35^{\circ}C$, 120 rpm for 40 h. With culture time, hydrogen production and cell growth increased, but residual glucose and pH decreased. When the $2\%$ of glucose was used as single carbon source, hydrogen production was 975.1 mL/L. To enhance hydrogen productivity, mixed carbon sources of glucose and sucrose were added. The maximum hydrogen production was earned at the mixing ratio of 25:75, and it was 1319.5 mL/L. When we added 50 mM of phosphate to protect the pH drop in culture broth, hydrogen production increased 1.3 times more than that of initial concentration. The organic nitrogen sources were more effective than inorganic nitrogen for hydrogen production. Among organic nitrogen, yeast extract was the most effective and its hydrogen production was 1691.3 mL/L. Among 9 of mineral sources, Ferric citrate and $NaMoO_4$ were especially effective, and their productions were 1782.3 mL/L and 1784.8 mL/L, respectively.

• Journal of Life Science
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• v.27 no.7
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• pp.805-811
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• 2017

In the previous paper, we isolated a bacterium that can hydrolyze various organic materials from soybean paste, including cellulose, lipids, starch, and protein. The activity and chemical properties of the crude enzymes produced by the isolate Bacillus subtilis CK-2 were further investigated. Cellulase showed the highest activity at pH 5.0 and $55^{\circ}C$. The stability of cellulase was maintained within the ranges of pH 5.0~10.0 and $20{\sim}50^{\circ}C$. Cellulolytic enzymes were activated by a $Co^{2+}$ ion, demonstrating the highest activity at a 0.45%(w/v) concentration of $Co^{2+}$. The optimal conditions for amylase were pH 5.0 and $50^{\circ}C$. The activity of amylase was stable within the ranges of pH 4.0~5.0 and $20{\sim}50^{\circ}C$. The $Co^{2+}$ ion was also necessary for amylase activity, which was the highest at a 0.2%(w/v) concentration of $Co^{2+}$. The optimal pH and temperature conditions of protease were pH 8.0 and $50^{\circ}C$. The activity of protease was stable within the ranges of pH 7.0~8.5 and $20{\sim}50^{\circ}C$. Protease activity was catalyzed by $Mn^{2+}$, which was the highest at a 0.125%(w/v) concentration of $Mn^{2+}$. The isolate B. subtilis CK-2 demonstrated a high activity of autolysin. Based on these results, we identified and suggested the optimal pH, temperature, and metal ion concentration in the use of the hydrolytic enzymes of B. subtilis CK-2 for industrial purposes.

• KSBB Journal
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• v.25 no.6
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• pp.520-526
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• 2010

In this work we have investigated the production of catalase from Bacillus sp. strains, which were screened and identified from soil. These strains were cultivated in shaking flasks with tryptic soy broth (TSB) at $30^{\circ}C$ and 200 rpm. Effects of the temperature and pH on the stability of the native catalase and whole cell viability were studied in the temperature range of $25-60^{\circ}C$ and the pH range of 7-13. Korean natural zeolite was added to culture medium and mixed with microorganisms for 24 hours. The native catalase maintained its activity over $50^{\circ}C$. The enzyme acitiviy of the catalase from Bacillus flexus BKBChE-3 was highest among the Bacillus sp. strains studied. Bacillus flexus BKBChE-3 and immobilized Bacillus cells have survived under extreme conditions of over $50^{\circ}C$ and pH 12. 60 mL of 10.5 mM $H_2O_2$ solution were entirely removed within 1 hour with catalase produced from Bacillus sp. on the flask. When Bacillus cells were immobilized on Korean natural zeolite, colony forming unit of Bacillus flexus BKBChE-3 was increased and high efficiency of hydrogen peroxide removal was observed.

• KSBB Journal
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• v.18 no.3
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• pp.186-189
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• 2003

This study aims to develop an economic process for the treatment of industrial wastewater containing hydrogen peroxide by using catalase. Core process is characterized by two membranes; microfiltration membrane and ultrafiltration membrane with different molecular cut-offs. Optimum dilution ratio of Aspergillus niger molds 개 buffer solution is 1:5. The final recovery yield of the enzyme is over 90% using the process. The enzyme solution shows the optimum temperature of 4$0^{\circ}C$ and pH range of 5-8.