Antioxidant and TNF-α Inducing Ability of Styela plicata Extracts

Hyun-Hyo Suh(서현효),Bo-Bae Lee(이보배),Seung-Cheol Lee(이승철) 한국생명과학회 2009 생명과학회지 Vol.19 No.4

메탄올, 에탄올, 아세톤 그리고 물을 이용하여 신선한 오만둥이의 추출물을 제조한 후, 추출물의 항산화능 및 TNF-α 유도능을 조사하였다. 아세톤 추출물을 제외한 대부분의 추출물은 malonedialdehyde 생성을 50% 이상 저해하는 항산화능을 보였으며, 메탄올 추출물은 가장 높은 값인 59.3%의 malonedialdehyde 생성 저해능을 나타내었다. 물과 아세톤 추출물은 각각 18.5와 12.3 μM의 glutathione 함량을 보였지만 그 외의 추출물에서는 검출되지 않았다. 추출물들의 TNF-α 유도능을 RAW 264.7 세포주로 조사하였다. 오만둥이 물 추출물은 첨가량이 증가할수록 TNF-α 생성을 증가시켰다. 그러나 그 이외의 추출물들은 RAW 264.7 세포주에서의 증가할수록 TNF-α 생성을 유도하지 못하였다. After preparing extracts from fresh Styela plicata using methanol, ethanol, acetone, and water, the antioxidant and tumor necrosis factor (TNF)-α inducing ability of the extracts were evaluated. Most of the extracts showed more than 50% inhibitory activity against malonedialdehyde formation, and methanol extract exhibited the highest activity with values of 59.3%. The water and acetone extracts contained glutathione contents of 18.5 and 12.3 μM, respectively, however, these were not detected in the other extracts. TNF-α inducing abilities of the extracts were determined with RAW 264.7 cells. The water extract increased TNF-α production with correlation to increase of the added amount. However, the other extracts could not induce TNF- α production in RAW 264.7 cells. The results indicated that the antioxidant activities of S. plicata extracts were different depending on extracting solvents, and the water extract showed significant TNF-α inducing activity.

Flocculating Properties of Bioflocculant Biopol32 from Pseudomonas sp. GP32

Hyun Don Lee(이현돈),Oh Nara(오나라),Muyeong Eun Lee(이명은),Hyun Hyo Suh(서현효) 한국생명과학회 2017 생명과학회지 Vol.27 No.8

Pseudomonas sp. GP32가 생산하는 생물고분자응집제 Biopol32의 실제 산업폐수에서의 적용을 위하여 생물고분자 Biopol32의 응집특성을 조사하였다. Biopol32의 응집물질은 polysaccharide로 확인되었다. 음이온성 응집제들은 응집효율을 높이기 위하여 보조응집제로 counter ion을 사용하고 있다. Biopol32의 응집활성은 보조응집제로 Ca<SUP>2+</SUP>, Al<SUP>3+</SUP>와 같은 양이온을 첨가 하였을 때 크게 증가하였으며, 보조응집제로서 7.0 mM CaCl₂ · 2H₂O을 첨가하였을 때 Biopol32의 응집활성이 가장 높게 나타났다. Kaolin 현탁액에 Biopol32를 1.5 mg/l의 농도로 첨가하였을 때 가장 높은 응집활성을 보였다. Biopol32의 pH와 온도에 따른 응집활성은 현재 폐수처리 현장에서 상업적으로 이용되고 있는 음이온성 유기합성고분자응집제 polyacrylamide와 Zoogloea ramigera로부터 생산된 생물고분자응집제 zooglan과 응집활성을 비교하였다. Biopol32의 응집활성은 pH 5.0에서 8.0의 넓은 범위의 pH에서 높은 응집활성을 보였으며, 또한 온도의 영향에서는 60℃에서 가장 높은 응집활성을 나타내었으나, 70℃ 이상의 온도에서는 응집활성이 급격히 감소하는 것으로 나타났다. Jar fermentor를 이용한 batch culture를 통하여 Biopol32의 응집활성과 전하밀도와의 관계를 조사하였다. 음이온성 전하밀도와 겉보기 점도가 높을수록 Biopol32의 응집활성이 높아져, Biopol32의 응집활성과 겉보기점도는 Biopol32의 전하밀도와 밀접한 관계가 있는 것으로 나타났다. The flocculating properties of bioflocculant Biopol32 produced by Pseudomonas sp. GP32 were investigated for application in industrial wastewater treatment. The major flocculating substance of bioflocculant Biopol32 was identified as polysaccharide. Many anionic flocculants need a counter ion to induce higher flocculating activity. The flocculating activity of bioflocculant Biopol32 was markedly increased by the addition of cationic ions (Ca<SUP>2+</SUP>, Al<SUP>3+</SUP>). The flocculating activity of bioflocculant Biopol32 was the most effective when 7.0 mM CaCl₂ · 2H₂O as coflocculant was added. The flocculating activity on the effect of pH and the temperature of the bioflocculant Biopol32 was compared with anionic commercial flocculant (polyacrylamide) and bioflocculant (zooglan from Zoogloea ramigera). In kaolin suspension, the highest flocculating activity was obtained at the bioflocculant Biopol32 concentration of 1.5 mg/l. A high flocculating activity was observed in the pH range of 5.0 to 8.0. The flocculating activity of bioflocculant Biopol32 was sustained up to 60℃, but decreased rapidly at over 70℃. In the batch culture, the charge density of bioflocculant Biopol32 was compared with flocculating activity. The larger the anionic charge density and apparent viscosity of bioflocculant Biopol32, the higher the flocculating activity. Therefore, we confirmed that the flocculating activity and apparent viscosity of bioflocculant Biopol32 was closely related to the charge density of bioflocculant Biopol32.

유기염소계 난분해성 산업폐수의 처리를 위한 미생물제제의 개발

이현돈(Hyun Don Lee),임성원(Seong Won Im),서현효(Hyun-Hyo Suh) 한국생명과학회 2014 생명과학회지 Vol.24 No.8

유기염소계 난분해성 산업폐수처리에 효과적인 미생물제제 개발을 위하여 PCP (pentachlorophenol)와 TCE (trichloroethylene) 등과 같은 유기계 염소화합물이 오염되어있는 토양 및 산업폐수로부터 PCP 분해활성이 높은 GP5, GP19와 TCE 분해활성이 높은 GA6, GA15를 분리하였다. 이들 분리균주 GP5, GP19, GA6, GA15등은 Acetobactor sp., Pseudomonas sp., Arthrobacer sp., Xanthomonas sp.과 유사한 것으로 나타나 최종적으로 Acetobacter sp. GP5, Pseudomonas sp. GP19, Arthrobacer sp. GA6, Xanthomoas sp. GA15로 명명하였다. 유기염소계 산업폐수의 처리를 위한 복합미생물제제 OC17은 PCP와 TCE를 분해하는 4개의 분리 분리균주와 방향족화합물 분해균주인 Acinetobacter sp. KN11, Neisseria sp. GN13의 배양액을 혼합하여 제조하였다. 복합미생물제제 OC17은 2.8×10<SUP>9</SUP> CFU/g의 균체수를 갖고 있으며, 밀도는 0.299 g/㎤, 수분함량은 26.8%를 나타내었다. 복합미생물 제제 OC17은 PCP 500 mg/l가 포함되어있는 인공폐수를 이용한 실험에서 배양 65시간에 87%의 분해효율을 나타내었고, TCE (300 uM)의 분해효율은 배양 50시간에 90%의 분해효율을 나타내었다. 복합미생물제제 OC17을 이용한 유기 염소계 산업폐수의 처리효율 시험을 위한 연속배양 실험 에서 10일간 처리 하였을 때 91%의 COD 제거효율을 나타내었다. The screening of the microorganisms degrading chlorinated organic compounds such as PCP (pentachlorophenol) and TCE (trichloroethylene) was conducted with soil and industrial wastewater contaminated with various chlorinated organic compounds. Isolates (GP5, GP19) capable of degrading PCP and isolates (GA6, GA15) capable of degrading TCE were identified as Acetobactor sp., Pseudomonas sp., Arthrobacer sp., Xanthomonas sp. and named Acetobacter sp. GP5, Pseudomonas sp. GP19, Arthrobacer sp. GA6 and Xanthomoas sp. GA15, respectively. The microbial augmentation, OC17 formulated with the mixture of bacteria including isolates (4 strains) degrading chlorinated organic compounds and isolates (Acinetobacter sp. KN11, Neisseria sp. GN13) degrading aromatic hydrocarbons. Characteristics of microbial augmentation OC-17 showed cell mass of 2.8×10<SUP>9</SUP> CFU/g, bulk density of 0.299 g/㎤ and water content of 26.8%. In the experiment with an artificial wastewater containing PCP (500 mg/l), degradation efficiency of the microbial augmentation OC17 was 87% during incubation of 65 hours. The degradation efficiency of TCE (300 uM) by microbial augmentation OC17 was 90% during incubation of 50 hours. In a continuous culture experiment, analysis of the biodegradation of organic compounds by microbial augmentation OC17 in industry wastewater containing chlorinated hydrocarbons showed that the removal rate of COD was 91% during incubation of 10 days. These results indicate that it is possible to apply the microbial augmentation OC17 to industrial wastewaters containing chlorinated organic compounds.

Phenol 분해균주의 분리 및 페놀함유 폐수의 생물학적 처리

이현돈(Hyun Don Lee),이명은(Myoung Eun Lee),김형갑(Hyung Gab Kim),서현효(Hyun-Hyo Suh) 한국생명과학회 2013 생명과학회지 Vol.23 No.10

방향족화합물들로 오염되어있는 토양 및 산업폐수를 포함한 각종 시료로부터 phenol에 분해활성이 높은 56균주를 순수분리 하였으며, 이들 분리 균주 중 균체생육과 phenol 분해활성이 가장 높은 균주인 GN13을 선별하였다. 분리균주 GN13은 형태학적, 생리학적 및 생화학적 특성을 조사한 결과 Neisseria 속 세균과 유사한 것으로 판명되어 최종적으로 Neisseria sp. GN13으로 명명하였다. 분리균주 Neisseria sp. GN13의 균체생육 및 phenol 분해를 위한 최적온도와 최적 pH는 각각 32℃와 7.0였다. 유일 탄소원으로 phenol 1,000 mg/l를 포함하여 최적화된 배지를 사용한 jar-fermentor 배지에서 배양 30시간에 균체생육이 최대에 이르렀으며 배양 27시간째 거의 모든 phenol이 분해되었으며, catechol deoxygenase 활성측정에 의하여 Neisseria sp. GN13은 meta-와 ortho-pathway를 통하여 catechol 분해가 일어났다. Neisseria sp. GN13은 phenol 함유 인공폐수에서의 phenol 분해율은 배양 30시간 만에 97%의 phenol이 분해되는 것으로 나타났으며, 인공폐수에 대한 Neisseria sp. GN13과 활성슬러지 처리구에서의 TOC 제거효율은 각각 83%와 78%였다. 석유화학폐수에 대한 Neisseria sp. GN13의 COD 제거율은 활성슬러지만을 포함한 대조구보다 약 1.3배 높은 효율을 나타내었다. 이러한 결과로 미루어 분리균주 Neisseria sp. GN13은 phenol을 함유하고 있는 여러 폐수에 효과적으로 적용될 수 있을 것으로 생각된다. Aromatic hydrocarbons, such as phenol, have been detected frequently in wastewater, soil, and groundwater because of the extensive use of oil products. Bacterial strains (56 isolates) that degraded phenol were isolated from soil and industrial wastewater contaminated with hydrocarbons. GN13, which showed the best cell growth and phenol degradation, was selected for further analysis. The GN13 isolate was identified as Neisseria sp. based on the results of morphological, physiological, and biochemical taxonomic analyses and designated as Neisseria sp. GN13. The optimum temperature and pH for phenol removal of Neisseria sp. GN13 was 32°C and 7.0, respectively. The highest cell growth occurred after cultivation for 30 hours in a jar fermentor using optimized medium containing 1,000 mg/l of phenol as the sole carbon source. Phenol was not detected after 27 hours of cultivation. Based on the analysis of catechol dioxygenase, it seemed that catechol was degraded through the meta- and ortho-cleavage pathway. Analysis of the biodegradation of phenol by Neisseria sp. GN13 in artificial wastewater containing phenol showed that the removal rate of phenol was 97% during incubation of 30 hours. The removal rate of total organic carbon (TOC) by Neisseria sp. GN13 and activated sludge was 83% and 78%, respectively. The COD removal rate by Neisseria sp. GN13 from petrochemical wastewater was about 1.3 times higher than that of a control containing only activated sludge.

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

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.

Pseudomonas fluorescens JK-0412의 키틴 분해 효소 및 단백질 분해효소 생산 최적화

박용일 ( Yong Il Park ),서현효 ( Hyun Hyo Suh ),김우중 ( Woo Jung Kim ),고주희 ( Ju Hee Ko ),박제권 ( Jae Kweon Park ) 한국키틴키토산학회 2010 한국키틴키토산학회지 Vol.15 No.4

Pseudomonas fluorescens JK-0412가 생산하는 키틴 분해 효소와 단백질 분해효소의 생산성을 다양한 기질을 이용하여 실험조사 하였다. 각각 키틴 분해효소와 단백질 분해효소 (CTNase와 PRTase로서 명명)의 생산에 여러 가지 기질의 특이성을 검토하였다. 전자는 주로, pNP-GlcNAc에 기질 특이성을 보이는 CTNase, 그리고, 후자는 skim-milk에 기질특이 성을 갖는 PRTase로서 두 효소의 최적 생산 조건은 각각 pH8.0와 37℃였으며, 또한 1.0% dextran, 1.0% yeast extract, 1.0% MgSO4를 각각 carbon, nitrogen, and mineral 원으로서 이용했을 때, CTNase의 최대 생산성을 얻을 수 있었다. 또한 각각, 1.0% glucose, 1.0% ammonium sulfate, 1.0% MgSO4를 carbon, nitrogen, 및 mineral 원으로서 이용했을 때, PRTase의 최대 생산성을 얻을 수 있었다. 특히, 유일한 carbon과 nitrogen 원으로 chitin powder, chitosan, colloidal chitin 과 dextran을 기질로 이용하여 12일 동안 배양하여 효소활성을 측정한 결과, CTNase 생산에 있어 기질로 powder chitin를 사용하였을 경우 3일 이후 효소활성이 확인되었으며, PRTase 활성이 측정되기 시작한 4일 이후의 PRTase 생산에 있어서도 powder chitin이 가장 적합한 기질로 확인되었다. 따라서 상업적으로 적용할 수 있는 chitin은 수산가공 부산물로부터 얻을 수 있을 수 생물소재로서 CTNase와 PRTase 생산을 위한 생물 공학적인 공정에 이용가치가 매우 높다. The production of chitinolytic and proteolytic enzymes in submerged cultures of Pseudomonas fluorescens JK-0412 in media was investigated in experiment with pH, temperature and substrate concentration as the main parameters. The production of chitinolytic and proteolytic enzymes, named as CTNase and PRTase, the former has specific enzyme activity toward pNPGlcNAc mostly and the latter has specific enzyme activity toward skim-milk, was monitored under the influence at various concentrations of these substrates. The optimum conditions of pH, temperature and substrate concentration for the production of both enzymes was determined to be pH 8.0 and 37 o C, and pH 8.0 and 45 o C, respectively. The maximum production of CTNase was obtained when 1.0% dextran, 1.0% yeast extract and 1.0% MgSO4 were used as carbon, nitrogen and mineral sources in the absence of chitinous material. In addition, the maximum production of PRTase was obtained when 1.0% glucose, 1.0% ammonium sulfate, and 1.0% MgSO4 were used as carbon, nitrogen and mineral source, respectively. Furthermore, in the presence of flake chitin, chitosan, and colloidal chitin used as a sole carbon and nitrogen source, it was found that flake chitin was the most efficient elicitor of CTNase and PRTase after 3 and 4 day cultivation, respectively. Hence chitin powder is a suitable substrate for the production of both CTNase and PRTase to employ in an advanced biotechnological process, whereby several commercially applicable chitinous materials can be obtained from a waste product of the fishing industry.

Biological Treatment of Recalcitrant Industrial Wastewater Using Microbial Augmentation and Bioflocculant-Producing Microorganisms

서상익 ( Sang-ik Suh ),박정호 ( Jung-ho Park ),서현효 ( Hyun-hyo Suh ) 한국환경기술학회 2024 한국환경기술학회지 Vol.25 No.1

For effective treatment of recalcitrant industrial wastewater, microbial augmentation J30, which is composed of Pseudomonas sp. GT21, Bacillus sp. KN27, Acinetobacter sp. KN11, and Neisseria sp. KN13, decomposing strains that show high activities on various organic substances and aromatic compounds, were prepared. As a method to control the solid suspension of industrial wastewater, the culture solutions of A and B, which produce bioflocculants, were applied to wastewater treatment. A microbial augmentation for treating laundry wastewater containing TCE and PCE were prepared by mixing 4 types of strains used to treat paper mill wastewater, 6 types of strains prepared by formulating culture solutions of Flavobacterium sp. GN18 and Acalgenes sp. GN23, which are richloroethylene and pentachloroethylene decomposing strains. It was named microbial augmentation TP32. These culture solutions of strain groups were added to treat paper mill wastewater containing phenolic chlorine compounds and laundry wastewater containing chlorine compounds efficiently. The removal efficiencies of the treatment groups, for TOC in paper mill wastewater were 81.9 % and 73.2 %, respectively. The TOC removal efficiency in paper mill wastewater was 86.5 % in the mixed strains treatment group, where decomposing and flocculating strains were added simultaneously. Changes in T-N and T-P showed a removal efficiency of more than 70 % in all treatments using decomposing, flocculating, and mixed strains. Still, the mixed strain treatment showed the highest removal efficiency. The phenolic chlorine compound, 3-chlorophenol removal efficiency was 85.1 % in the mixed strain treatment group. The removal efficiency of TCE from laundry wastewater showed a removal rate of 85.3 % in 42 hours of incubation when only decomposing strains were added and a removal rate of more than 90 % in 32 hours of incubation when mixed strains were added. The PCE removal rate has been decreased over time of incubation in the decomposing strain treatment and mixed strain treatment, respectively. For effective treatment of recalcitrant industrial wastewater, microbial augmentation J30, which is composed of Pseudomonas sp. GT21, Bacillus sp. KN27, Acinetobacter sp. KN11, and Neisseria sp. KN13, decomposing strains that show high activities on various organic substances and aromatic compounds, were prepared. As a method to control the solid suspension of industrial wastewater, the culture solutions of A and B, which produce bioflocculants, were applied to wastewater treatment. A microbial augmentation for treating laundry wastewater containing TCE and PCE were prepared by mixing 4 types of strains used to treat paper mill wastewater, 6 types of strains prepared by formulating culture solutions of Flavobacterium sp. GN18 and Acalgenes sp. GN23, which are richloroethylene and pentachloroethylene decomposing strains. It was named microbial augmentation TP32. These culture solutions of strain groups were added to treat paper mill wastewater containing phenolic chlorine compounds and laundry wastewater containing chlorine compounds efficiently. The removal efficiencies of the treatment groups, for TOC in paper mill wastewater were 81.9 % and 73.2 %, respectively. The TOC removal efficiency in paper mill wastewater was 86.5 % in the mixed strains treatment group, where decomposing and flocculating strains were added simultaneously. Changes in T-N and T-P showed a removal efficiency of more than 70 % in all treatments using decomposing, flocculating, and mixed strains. Still, the mixed strain treatment showed the highest removal efficiency. The phenolic chlorine compound, 3-chlorophenol removal efficiency was 85.1 % in the mixed strain treatment group. The removal efficiency of TCE from laundry wastewater showed a removal rate of 85.3 % in 42 hours of incubation when only decomposing strains were added and a removal rate of more than 90 % in 32 hours of incubation when mixed strains were added. The PCE removal rate has been decreased over time of incubation in the decomposing strain treatment and mixed strain treatment, respectively.

Isolation and Immunomodulating Activity of an Extracellular Polysaccharide Produced by Bacillus sp. PS-12

Ye Seul Na(나예슬),Hyun Hyo Suh(서현효) 한국생명과학회 2009 생명과학회지 Vol.19 No.6

토양에서 분리한 세포외 다당류 생산균주 PS-12는 형태학적, 생리학적, 화학적 분석에 의하여 Bacillus sp.에 속하는 균주로 동정되었으며, 분리균주 PS-12는 Bacillus sp. PS-12로 명명하였다. Bacillus sp. PS-12가 생산하는 세포외 다당류는 에탄올 침전, cetylpyridinium chloride (CPC) 침전과 gel permeation chromatography를 이용하여 정제하였으며, 정제된 다당류 PS-12의 당조성은 glucose, mannose, galactose와 fucose가 7:3.2:2:1의 몰비로 구성되어있었다. Bacillus sp. PS-12로부터 분리된 다당류 PS-12를 이용하여 면역증강효과를 확인하였다. TNF-α 및 IL-6 측정은 RAW264.7 대식세포주를 사용하였으며 cytokine정량을 위하여 ELISA kit를 이용하였다. RAW264.7 세포주에 대한 PS-12의 세포독성을 확인하기 위하여 세포독성이 10% 미만을 나타내는 농도인 2 ㎍/㎖을 PS-12의 최대농도로 측정하였다. PS-12는 2 ㎍/㎖에서 TNF-α를 정상세포보다 50배 이상 높은 수치로 생산하였다. 또한 IL-6의 생산을 농도 의존적으로 증가시켰다. 이러한 결과로부터 PS-12가 면역세포에 대해 세포독성을 거의 나타내지 않는 농도에서 대식세포로부터 TNF-α와 IL-6의 cytokine 생산을 함으로써 면역증강효과를 나타낸다는 것을 확인하였다. A bacterial strain producing highly viscous extracellular polysaccharide was isolated from soil. Through morphological, physiological and chemotaxonomical studies, it was identified as a Bacillus sp. and named Bacillus sp. PS-12. The extracellular polysaccharide, named PS-12 was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation and gel permeation chromatography. The purified polysaccharide was found to consist of glucose, mannose, galactose, and fucose, with a molar ratio of approximately 7:3.2:2:1, respectively. PS-12 was investigated for its immunostimulating activity on murine macrophage RAW264.7 cells using an ELISA assay. PS-12 stimulated the production of TNF-α to a level 50 times greater than the control and also induced IL-6 secretion in a dose-dependent manner. The cytotoxicity on RAW264.7 cells by PS-12 was relatively low with 10% cytotoxicity at 2 ㎍/㎖. These results indicate that PS-12 is less cytotoxic to immune cells and possess immunomodulating activity in which it can produce cytokines including TNF-α and IL-6 from macrophages.

제지폐수 처리용 미생물의 분리 및 복합 미생물제제의 개발

Dae Ook Kang(강대욱),Hyun-Hyo Suh(서현효) 한국생명과학회 2011 생명과학회지 Vol.21 No.4

제지폐수의 효율적인 생물학적 처리와 폐수특성에 적합한 미생물제제의 개발을 위하여 토양 및 산업폐수로부터 방향족 화합물에 분해활성이 높은 KN11, KN13 및 KN27 균주와 세포 외 섬유소 가수분해효소 생산 균주 GT21 등의 균주를 분리하였다. 형태학적, 생리학적 및 생화학적 분류를 통해 이들 분리주 KN11, KN13, KN27 및 GT21 등은 Acinetobactersp., Neisseriasp., Bacillussp., Pseudomonassp.와 유사한 것으로 판명되어 최종적으로 각각 Acinetobactersp. KN11, Neisseriasp. KN13, Bacillussp. KN27, Pseudomonassp. GT21로 명명하였다. 제지폐수 중 난분해성 물질과 COD 증가원인 물질을 분석하고자 GC/MS를 이용하여 방향족 화합물 및 그 유도체들을 검출하였다. 분리균주 Acinetobactersp. KN11, Neisseriasp. KN13, Bacillussp. KN27 및 Pseudomonassp. GT21의 균체로 구성된 미생물제제 J30을 제조하여 제지폐수의 효율적 처리를 위한 연구에 사용하였다. 미생물제제 J30의 제지폐수에서 COD 제거를 위한 최적온도와 pH는 각각 30℃와 7.5였으며 배양 60시간에서 최대의 COD 제거효율을 나타내었다. 실험실 규모의 pilot plant에서 미생물제제 J30의 COD 제거효율은 87%의 높은 제거효율을 나타내었다. This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobactersp., Neisseriasp., Bacillussp., and Pseudomonassp. and named Acinetobactersp. KN11, Neisseriasp. KN13, Bacillussp. KN27, and Pseudomonassp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobactersp. KN11, Neisseriasp. KN13, Bacillussp. KN27, and Pseudomonassp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were 30℃ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.