원유 분해균주 Nocardia sp. H17-1의 분리 및 특성

이창호(Chang-Ho Lee),권기석(Gi-Seok Kwon),김희식(Hee-Sik Kim),서현효(Hyun-Hyo Suh),안극현(Keuk-Hyun Ahn),오희목(Hee-Mock Oh),권태종(Tae-Jong Kwon),윤병대(Byung-Dae Yoon) 한국생물공학회 1996 KSBB Journal Vol.11 No.6

식품 및 환경 , 기타 Acinetobacter sp . A54 에 의한 Arabian Light 원유의 분해

이창호,김희식,서현효,최성훈,오희목,윤병대 ( Chang Ho Lee,Hee Sik Kim,Hyun Hyo Suh,Soung Hun Choi,Hee Mock Oh,Byung Dae Yoon ) 한국미생물생명공학회 1997 한국미생물·생명공학회지 Vol.25 No.5

Biodegradation of Benzne,Toluene, and Phenol by a Mixed Culture in Semicontinuous Culture

오희목,김성빈,이창호,서현효,이문호,고영희,윤병대,Oh, Hee-Mock,Kim, Seong-Bin,Lee, Chang-Ho,Suh, Hyun-Hyo,Lee, Moon-Ho,Kho, Yung-Hee,Yoon, Byung-Dae 한국미생물 · 생명공학회 1994 한국미생물·생명공학회지 Vol.22 No.4

The biodegradation of aromatic compounds by a mixed culture GE1 was investigated in an artificial wastewater containing 250 mg/l of benzene, toluene, and phenol in semicontinuous culture. In the control group (no strains) with an aeration rate of 75 ml/l/min, 37% of phenol and 83% of benzene were volatilized during early 24 hrs and toluene was disappeared from the medium within 12 hrs. The biodegradation of benzene and toluene was effective in SB (strains + biofilm) treatment, while phenol was degraded more quickly in SG (strains + glucose) treatment including glucose as an additional carbon source. aromatic compounds added at a concentration of 250 mg/l were completely removed by SG treatment after 16 hrs or 32 hrs, respectively. The removal rate of COD was high as much as 80 mg/l/h in SG treatment during early period, but COD revealed a stable value of 116~140 mg/l after 12 hrs caused by increased biomass. Therefore, it is concluded that the mixed GE1 could be used for the wastewater treatment including aromatic compounds such as benzene, toluene, and phenol.

알칼리성 Prottease를 생산하는 Xanthomonas sp. YL-37의 분리 및 조효소의 성질

이창호,권태종,강상모,서현효,권기석,오희목,윤병대,Lee, Chang-Ho,Kwon, Tae-Jong,Kang, Sang-Mo,Suh, Hyun-Hyo,Kwon, Gi-Seok,Oh, Hee-Mock,Yoon, Byung-Dae 한국미생물 · 생명공학회 1994 한국미생물·생명공학회지 Vol.22 No.5

A bacterial strain, which showed the high protease activity at low temperature and the high tolerance for the surfactant, was isolated from soil and identified as Xanthomonas sp. YL-37. The optimal temperature, initial pH, and cultivation time for the production of the alkaline protease by Xanthomonas sp. YL-37 were 20$\circC , 11.0, and 84 hours, respectively. In the jar fermenter culture of Xanthomonas sp. YL-37, the alkaline protease activity was about 15,000 DU/ml/-broth after cultivating for 108 hours. The optimal pH and temperature for the protease activity were 70$\circC and 11.0, respectively. The protease was relatively stable at the pH range of 7.0~12.0 and at the temperatures below 50$\circC . The protease activity at 20$\circC was about the level of 40% of its activity at 70$\circC . The enzyme was suggested as a serine protease because the enzyme activity was inhibited by phenylmethane sulfonyl fluoride, a serine modifier.

페놀분해 효모 Candida tropicalis PW-51의 분리 및 분해특성

김성빈,김치경,김희식,이창호,신기선,권시석,윤병대,오희목 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.6

본 연구는 페놀과 포름알데히드를 포함하는 페놀계수지 산업폐수의 생물학적 처리에 이용할 목적으로 포름알데히드의 존재하에서 페놀분해능이 우수한 효모를 sludge로부터 분리하여, 형태적 및 생리 생화학적 특징을 조사하여 Candida tropicalis PW-51로 동정하였다. 회분식 배양에서 C. tropicalis PW-51의 페놀 분해한계는 2,000 mg/ℓ이며, 58시간내에 완전히 분해하였다. C. tropicalis PW-51은 초기접종량이 9×10 exp (6) cells/㎖, 배양온도는 30℃, pH는 7.0에서 페놀 분해효율이 높았으며, 500~2,000 mg/ℓ의 페놀농도에서 조사된 페놀에 대한 평균 분해율은 페놀 1,000 mg/ℓ에서 45.5 mg/ℓ/h 가장 높았다. C. tropicalis PW-51의 페놀분해시 catechol 1,2-dioxygenase의 활성이 크게증가하므로 ortho-path-way에 의해 페놀을 분해하는 것으로 판단되었다. 따라서, C. tropicalis PW-51는 페놀계 수지 산업폐수의 생물학적 처리에 효과적으로 이용될 수 있는 균주로 사료된다. For the biological treatment of phenolic resin wastewater containing phenol and formaldehyde, a phenol-degrading yeast was isolated from the papermill sludge, and then identified as Candida tropicalis PW-51 according to morphological, physiological and biochemical properties. The strain was able to degrade high phenol concentrations up to 2,000 mg/ℓ within 58 hours in batch cultures. Phenol-degrading efficiency by the strain was maximum at the culture conditions of a final concentration of 9×10 exp (6) cells/㎖ 30℃ and pH 7.0. The mean degradation rate of phenol was highest at 45.5 mg/ℓ/h in 1,000 mg/ℓ phenol from 500 mg/ℓ to 2,000 mg/ℓ phenol. Because the enzyme activity of catechol 1,2-dioxygenase increased in the course of degradation of phenol, it seems that this strain degrades phenol via the ortho-cleavage of benzene ring. The isolate C. tropicalis PW-51 could be effectively used for the biological treatment of phenolic resin wastewater.

알칼리성 Protease를 생산하는 Xanthomonas sp. YL-37의 분리 및 조효소의 성질

이창호,권태종,강상모,서현효,권기석,오희목,윤병대 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.5

저온에서 높은활성을 나타내며, 계면활성제에 내성이 있는 알칼리성 protease를 생산하는 균주를 토양으로부터 분리·선발하였으며, Xanthomonas sp. YL-37로 명명하였다. 효소생산을 위한 배양 최적온도, 초발 pH, 그리고 배양시간은 각각 20℃, 11.0, 그리고 84시간이였다. Jar fermenter 배양을 했을 때 배양 후 108시간에 효소의 역가는 약 15,000 DU/㎖-broth였다. 본 알칼리성 protease의 최적온도 및 pH는 70℃ 및 10.0이였으며, 20℃ 에서도 최적활성의 약 40%를 유지하였다. PH 및 열안정성은 각각 pH 7.0∼12.0, 온도 50℃ 이하에서 비교적 안정하였다. 또한 Xanthomonas sp. YL-37이 생산하는 protease는 PMSF에 강하게 저해를 받아서 본 효소는 활성부위에 serine 잔기를 가지고 있는 일종의 serine protease로 사료된다. A bacterial strain, which showed the high protease activity at low temperature and the high tolerance for the surfactant, was isolated from soil and identified as Xanthomonas sp. YL-37. The optimal temperature, initial pH, and cultivation time for the production of the alkaline protease by Xanthomonas sp. YL-37 were 20℃, 11.0, and 84 hours, respectively. In the jar fermenter culture of Xanthomonas sp. YL-37, the alkaline protease activity was about 15,000 DU/㎖-broth after cultivating for 108 hours. The optimal pH and temperature for the protease activity were 70℃ and 11.0, respectively. The protease was relatively stable at the pH range of 7.0∼12.0 and at the temperatures below 50℃. The protease activity at 20℃ was about the level of 40% of its activity at 70℃. The enzyme was suggested as a serine protease because the enzyme activity was inhibited by phenylmethane sulfonyl fluoride, a serine modifier.

Phenol을 분해하는 Acinetobacter sp. GEM2의 분리 및 특성

이창호,오희목,권태종,권기석,이성기,서현효,윤병대 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.6

Phenol을 증기상으로 공급하여 배양하면서 colony 형성이 우수하고, 액체배양에 의한 균생육과 분해능이 우수한 균주를 선발하여 Acinetobacter sp. GEM2로 동정하였다. Acinetobacter sp. GEM2의 생육을 위한 최적온도 및 초발 pH는 30℃, 7.0이였으며, phenol 1500 ppm 이상의 농도에서는 기질저해를 나타내었다. 배양 10시간 이후에 급속한 균생육을 보여 배양 후 24시간에는 거의 정지기에 도달하였으며, phenol 농도의 95% 이상이 분해되었다. Acinetobactoer sp. GEM2는 benzoic acid, phenol, m-cresol, o-cresol, p-cresol, catechol, gentisic acid 그리고 toluene 등의 방향족 화합물을 이용하며, benzene, salicylic acid, p-toluic acid 그리고 p-xylene은 전혀 이용하지 못하였다. Acinetobacter sp. GEM2는 ortho-와 meta-분해경로를 통하여 catechol을 분해하며, 유기용매에 대한 성장제한 log P 값은 2.0 정도였다. A bacterial strain which formed a distinct colony on agar plate containing phenol as a vapor phase and grew well in a liquid minimal medium was isolated and identified as Acinetobacter sp. GEM2. The optimal temperature and initial pH for the growth of Acinetobacter sp. GEM2 were 30℃ and 7.0, respectively. Cell growth was inhibited by phenol at the concentration over 1500 ppm. Cell growth dramatically increased from 10 hours after cultivation and almost showed a stationary phase within 24 hours at which 95% of phenol was concomitantly degraded. Acinetobacter sp. GEM2 was capable of growing on aromatic compounds, such as benzoic acid, phenol, m-cresol, ο-cresol, p-cresol, catechol, gentisic acid, and toluene, but did not grow on benzene, salicylic acid p-toluic acid, and p-xylene. By the analysis of catechol dioxygenase, it seemed that catechol was degraded through both meta- and ortho-cleavage pathway. The growth-limiting log P value of Acinetobacter sp. GEM2 on organic solvents was 2.0.

단일배양 및 혼합배양에 의한 Benzene, Phenol 및 Toluene 혼합물의 생분해

이창호,오희목,권태종,권기석,김성빈,고영희,윤병대 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.6

Benzene(B), phenol(P), 그리고 toluene(T)을 각각 500 ㎎/ℓ 포함한 합성폐수에 분리·선별된 단일균주 및 혼합균주를 접종하고 배양하면서 기질 조합에 따른 방향족화합물의 생분해에 대하여 조사하였다. 사용된 세 균주 모두가 탄소원으로서 p-xylene을 이용하지 못하였으나 benzene 또는 toluene과 함께 p-xylene을 첨가할 경우에는 균생육이 높았다. 기질조합에 의한 혼합배양시 유도기는 혼합물의 종류에 따라 서로 차이가 있었다. Flavobacterium sp. BEN2와 Acinetobacter sp. GEM63의 균생육은 500㎎/ℓ의 phenol이 존재할 경우 저해를 받았다. 반응조에서 합성폐수를 이용한 세 균주의 혼합배양을 실시하였을 때, 처리구에서 benzene, phenol, 그리고 toluene은 배양 후 각각 30시간, 50시간, 12시간에 검출되지 않았으며, COD_t의 제거율은 배양 50시간에 약 80%를 나타내었다. 또한 배양액을 원심분리한 후 상징액의 COD_s는 80㎎/ℓ로서 93.8%의 COD 제거율을 나타내었다. The biodegradation of aromatic compounds by mixed and monoculture was investigated in an artificial wastewater containing 500 ㎎/ℓ of benzene(B), phenol(P), and toluene(T) in various combinations. None of three strains utilized p-xylene(X) as a carbon source, but they grew well on p-xylene in mixtures with benzene and toluene. In the mixed culture on mixed substrate, the length of lag phase was different depending on the nature of mixture. Cell growths of Flavobacterium sp. BEN2 and Acinetobacter sp. GEM63 were inhibited in the presence of a 500 ㎎/ℓ of phenol. When the mixed culture of three strains was cultured in a bench-scale reactor containing artificial wastewater, each of benzene, phenol, and toluene was not detected at 30 hrs, 50 hrs, and 12 hrs after incubation in the treatment. The removal rates of COD_t(total COD) and COD_s(soluble COD) of upper phase after centrifugation during early 50 hrs were ca. 80% and ca. 93.8%, respectively.

Naphthalene 분해균주 Alcaligenes sp. A111 분리 및 특성

오희목,강정현,이창호,박찬선,안성구,윤병대,고영희 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.4

Naphthalene을 증기상으로 공급하고 배양하면서 세균집락 형성이 우수하고, 액체배양에 의한 균체증식이 탁월한 균주를 선별하여 Alcaligenes sp. A111로 동정하였다. Alcaligenes sp. A111의 생육을 위한 최적 온도는 30℃, 최적 pH는 7.0이며, 배양 후 12시간부터 급속한 균생장을 보여 배양 48시간에는 거의 정지기에 도달하였다. Naphthalene 농도에 따른 상대생장율은 영양염류의 농도에 대한 전형적인 미생물생장의 유형을 따르는 것으로서, 최대 상대생장율은 2.8/day이며 500∼4000 ppm의 농도에서 더 이상의 증가가 없었다. 무기질소원으로는 NH_4Cl과 NH_4NO_3에서 그리고 인과 질소의 질량비는 약 6 : 1에서 균생장이 우수하였다. Alcaligenes sp. A111는 catechol, gentisic acid, salicylic acid 등을 효과적으로 이용하며, benzene, toluene 및 octane에 대하여 내성을 보임으로서 이들 물질이 naphthalene과 혼합된 산업폐수의 처리에 효과적으로 이용될 수 있는 균주로 판단되었다. A bacterial strain which formed a distinct colony on agar plate containing naphthalene as a vapor phase and grew well in a liquid minimal medium was isolated and identified as Alcaligenes sp. A111. Optimum temperature and pH for the cultivation of Alcaligenes sp. A111 were 30℃ and 7.0, respectively. Cell growth increased dramatically from 12 hours after inoculation and revealed a stationary phase at about 48 hours. Relative growth rate (μ’) increased hyperbolically depending on the concentration of naphthalene up to 500 pm and reached to the maximum value of 2.8/day, but μ’ didn’t change within a range of 500∼4000 ppm naphthalene. NH_4Cl or NH_4NO_3 was preferred as a nitrogen source and a P : N ratio by weight of 6 : 1 was favorable to cell growth. Alcaligenes sp. A111 utilized the intermediates of degradation of naphthalene and showed tolerance to benzene, toluene, and octane. Therefore, it is suggested that Alcaligenes sp. A111 could be effectively used for the biological treatment of wastewater containing naphthalene in the presence of some aromatic compounds.

Bacillus sp. A29에 의한 다당류의 생산과 물성

안성구,서현효,이창호,오희목,권기석,이동희,윤병대 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.2

높은 점도를 보이는 다당류를 생산하는 균주 A-29를 선별하여 특성을 조사한 결과 Bacillus sp.으로 동정되었다. 배지 조성을 dextrin 12%, soytone 0.2%, SnCl_2 0.02%, Na_2HPO_4·12H_2O 0.36%, L-alanine 0.01%으로 하고 초발 pH를 6.8으로 하여 30℃ 에서 4일간 배양하였을 때 배양액의 점도는 65,000 cp였다. Jar fermentor 배양에서 생산된 A29 POL의 양은 8.3 g/ℓ이었으며, A29 POL의 단당류 조성은 glucose와 xylose가 8 : 1의 mole 비율로 구성되어 있었다. A29 POL은 0.1%의 낮은 농도에서도 높은 점성를 나타냈고 의가소성의 성질을 보였다. 특히 NaCl과 CaCl_2 등의 염에 의하여 크게 영향을 받으며 0.1%의 염 첨가시 점도가 급격히 증가하였고 의가소성도 더욱 증가하였다. 첨가된 염 농도가 0.3에서 0.6%까지 증가하였을 때, 완만한 감소를 보였다. 또한 높은 온도에서는 점도가 낮아졌지만 온도를 낮추었을 때는 점도가 다시 회복되는 현상을 보였다. 산성의 영역에서 높은 점도를 보였고 pH 3에서 가장 높았다. 알칼리성에서 낮은 점도를 보였다. A29 POL은 anionic polysaccharide이며 film을 형성할 수 있었다. MgSO_4·7H_2O, Na_2CO_3·H_2O, MnSO_4·7H_2O 등의 금속이온을 첨가하였을 때 gel을 형성하였다. A bacterial strain producing highly viscous polysaccharide(A29 POL) was isolated from soil and identified as Bacillus sp. A29. The cultural conditions of the Bacillus sp. A29 for the polysaccharide production were dextrin 12%, soytone 0.2%, SnCl_2·2H_2O 0.02%, Na_2HPO_4·12H_2O 0.36%, L-alanine 0.01%, initial pH 6.8, and 30℃ at pH 3 for 4 days. Final viscosity of the culture broth was 65,000 cp and then the amount of produced polysaccharide was 8.3 g/ℓ. A29 POL was composed of glucose and xylose. A29 POL showed high viscosity at low concentration(0.1%) and in the presence of the salts such as NaCl or CaCl_2. A29 POL showed high viscosity acid condition and at alkali condition and high pseudoplasticity in the presence of NaCl or CaCl_2. It was shown that the viscosity at high temperature(80℃) was decreased but it was recovered at low temperature(20℃). A29 POL was able to form film and gel in the presence of MgSO_4·7H_2O, Na_2CO_3·H_2O, MnSO_4·7H_2O. A29 POL had anionic charge.