가금폐기물 처리를 위한 세균유래 케라틴 분해효소의 특성

고태훈 ( Tae Hun Go ),이상미 ( Sang Mee Lee ),조광식 ( Kwang Sik Cho ),이예람 ( Ye Ram Lee ),박수연 ( Soo Yun Park ),장은영 ( Eun Young Jang ),정성윤 ( Seong Yun Jeong ),손홍주 ( Hong Joo Son ) 한국환경과학회 2014 한국환경과학회지 Vol.23 No.6

Keratin wastes are generated in excess of million tons per year worldwide and biodegradation of keratin by microorganisms possessing keratinase activity can be used as an alternative tool to prevent environmental pollution. For practical use of keratinase, its physicochemical properties should be investigated in detail. In this study, we investigated characteristics of keratinase produced by Xanthomonas sp. P5 which is isolated from rhizospheric soil of soybean. The level of keratinase produced by the strain P5 increased with time and reached its maximum (10.6 U/ml) at 3 days. The production of soluble protein had the same tendency as the production of keratinase. Optimal temperature and pH of keratinase were 40℃-45℃ and pH 9, respectively. The enzyme showed broad temperature and pH stabilities. Thermostability profile showed that the enzyme retained 94.6%-100% of the original activity after 1 h treatment at 10℃-40℃. After treatment for 1 h at pH 6-10, 89.2%-100% of the activity was remained. At pH 11, 71.6% of the original activity was retained after 1 h treatment. Although the strain P5 did not degrade human hair, it degraded duck feather and chicken feather. These results indicate that keratinase from Xanthomonas sp. P5 could be not only used to upgrade the nutritional value of feather hydrolysate but also useful in situ biodegradation of feather.

난분해성 케라틴 폐기물 처리를 위한 우모 분해 미생물의 분리 및 특성

고태훈 ( Tae Hun Go ),정진하 ( Jin Ha Jeong ),이나리 ( Na Ri Lee ),정성윤 ( Seong Yun Jeong ),박근태 ( Geun Tae Park ),손홍주 ( Hong Joo Son ) 한국환경과학회 2012 한국환경과학회지 Vol.21 No.2

We isolated and characterized novel duck feather-degrading bacteria producing keratinase. Twelve strains were isolated from soil and faces at poultry farm, and decayed feathers. They were identified as Bacillus methylotrophicus, Pseudomonas geniculata, Pseudomonas hibiscicola, Exiquobacterium profundum, Bacillus pumilus, Bacillus amyloliquefaciens, Chryseobacterium indologenes, Bacillus thuringiensis, Thermomonas koreensis, respectively, by phenotypic characters and 16S rRNA gene analysis. Generally, the level of keratinase production was not proportional to feather degradation rate. The highest keratinolytic activity was observed in the culture inoculated with Chryseobacterium indologenes D27. Although all strains did not degrade human hair, strains tested effectively degraded chicken feather(53.8-91.4%), wool(40.4-93.0%) and human nail (51.0-82.9%). These results suggest that strains isolated could be not only used to improve the nutritional value of recalcitrant feather waste but also is a potential candidate for biotechnological processes of keratin hydrolysis.

난분해성 우모분해 Pseudomonas geniculata에 의한 케라틴 분해효소 생산 및 식물성장 촉진 활성

고태훈 ( Tae Hun Go ),이상미 ( Sang Mee Lee ),이나리 ( Na Ri Lee ),정성윤 ( Seong Yun Jeong ),홍창오 ( Chang Oh Hong ),손홍주 ( Hong Joo Son ) 한국환경과학회 2013 한국환경과학회지 Vol.22 No.11

We investigated the optimal conditions for keratinase production by feather-degrading Pseudomonas geniculata H10 using one variable at a time (OVT) method. The optimal medium composition and cultural condition for keratinase production were determined to be glucose 0.15% (w/v), beef extract 0.08% (w/v), KH2PO4 0.12% (w/v), K2HPO4 0.02% (w/v), NaCl 0.07% (w/v), MgSO4;7H2O 0.03%, MgCl2;6H2O 0.04% along with initial pH 10 at 200 rpm and 25줛, respectively. The production yield of keratinase was 31.6 U/ml in an optimal condition, showing 4.6-fold higher than that in basal medium. The strain H10 also showed plant growth promoting activities. This strain had ammonification activity and produced indoleacetic acid (IAA), siderophore and a variety of hydrolytic enzymes such as protease, lipase and chitinase. Therefore, this study showed that P. geniculata H10 could be not only used to upgrade the nutritional value of feather wastes but also useful in situ biodegradation of feather wastes. Moreover, it is also a potential candidate for the development of biofertilizing agent applicable to crop plant soil.

Bacillus amyloliquefaciens를 이용하여 콩 종류와 발효온도를 달리하여 제조한 청국장의 특성

이나리,고태훈,이상미,홍창오,박규민,박근태,황대연,손홍주,Lee, Na-Ri,Go, Tae-Hun,Lee, Sang-Mee,Hong, Chang-Oh,Park, Kyu-Min,Park, Geun-Tae,Hwang, Dae-Youn,Son, Hong-Joo 한국미생물학회 2013 미생물학회지 Vol.49 No.1

This study was carried out to investigate the characteristics of Chungkookjang depending on different soybeans and fermentation temperatures using Bacillus amyloliquefaciens C2 isolated from homemade Chungkookjang. The highest protease activity was 854 U/g in yellow soybean Chungkookjang and 847 U/g in black soybean Chungkookjang at $35^{\circ}C$. The highest amylase activity was 3.87 U/g at $40^{\circ}C$ in yellow soybean Chungkookjang and 4.96 U/g at $45^{\circ}C$ in black soybean Chungkookjang. The highest reducing sugar content was 16.11 mg/g at $40^{\circ}C$ in yellow soybean Chungkookjang and 19.08 mg/g at $45^{\circ}C$ in black soybean Chungkookjang. The highest amino type nitrogen content was 420 mg%/g in yellow soybean Chungkookjang and 194 mg%/g in black soybean Chungkookjang at $40^{\circ}C$. The highest pH was 7.92 at $40^{\circ}C$ in yellow soybean Chungkookjang and 7.59 at $45^{\circ}C$ in black soybean Chungkookjang. The highest number of viable cell was 9.3 log CFU/g at $40^{\circ}C$ in yellow soybean Chungkookjang and at $35^{\circ}C$ in black soybean Chungkookjang. On the other hand, the lowest ammonia type nitrogen content was 225 mg%/g at $45^{\circ}C$ in yellow soybean Chungkookjang and 80 mg%/g at $40^{\circ}C$ in black soybean Chungkookjang. Yellow soybean Chungkookjang showed high protease activity, pH and amino type nitrogen, whereas black soybean Chungkookjang showed high amylase activity and reducing sugar. 본 연구에서는 가정에서 제조한 청국장으로부터 순수분리한 Bacillus amyloliquefaciens C2를 이용하여 콩 종류와 발효온도에 따른 청국장의 품질 특성 변화를 조사하였다. Protease 활성은 $35^{\circ}C$에서 배양했을 때 황태 청국장에서 854 U/g, 흑태 청국장에서 847 U/g로 가장 높았다. Amylase 활성은 황태 청국장의 경우, $40^{\circ}C$에서 3.87 U/g, 흑태 청국장은 $45^{\cricr}C$에서 4.96 U/g로 가장 높았다. 환원당 함량은 황태 청국장의 경우, $40^{\circ}C$에서 16.11 mg/g, 흑태 청국장은 $45^{\circ}C$에서 19.08 mg/g으로 가장 높았다. 아미노태 질소 함량의 경우, 황태 청국장은 $40^{\circ}C$에서 420 mg%/g, 흑태 청국장은 194 mg%/g로 가장 높았다. pH의 경우, 황태 청국장은 $40^{\circ}C$에서 7.92, 흑태 청국장은 $45^{\circ}C$에서 7.59로 가장 높았다. 황태 및 흑태 청국장의 생균수는 각각 $40^{\circ}C$ 및 $35^{\circ}C$에서 9.3 log CFU/g로 가장 높았다. 반면에 암모니아태 질소 함량은 황태 청국장의 경우, $45^{\circ}C$에서 225 mg%/g, 흑태 청국장은 $40^{\circ}C$에서 80 mg%/g로 가장 낮았다. 황태 청국장은 높은protease 활성, pH 및 아미노태 질소함량을 나타내었다. 반면에, 흑태 청국장은 높은 amylase 활성과 환원당 함량 및 낮은 암모니아태 질소 함량을 나타내었다.

터널 공동구 및 배수구에 사용되는 콘크리트 덮개 적용 사례 - 자동생산 시스템을 통한 미끄럼 방지 콘크리트 뚜껑 제품 -

강태성,고태훈,Gang, Tae-Seong,Go, Tae-Hun 한국터널지하공간학회 2017 자연, 터널 그리고 지하공간 Vol.19 No.3

Bacillus subtilis에 의한 발아 및 미발아 황태 청국장 발효

이나리 ( Na Ri Lee ),고태훈 ( Tae Hun Go ),박성보 ( Sung Bo Park ),이상미 ( Sang Mee Lee ),황대연 ( Dae Youn Hwang ),김동섭 ( Dong Seob Kim ),박근태 ( Geun Tae Park ),손홍주 ( Hong Joo Son ) 한국미생물생명공학회(구 한국산업미생물학회) 2013 한국미생물·생명공학회지 Vol.41 No.2

요약하면, 콩은 6시간 침지 및 2시간마다 4일 동안 물 공급 및 25oC에서 발아율이 가장 높았다. Isoflavone 함량은 발아 전보다 발아 후에 높게 나타났다. 발아콩-B. subtilis D7을 이용하여 발효된 청국장의 경우 아미노태 질소 함량, protease 활성 및 α-amylase 활성 등이 미발아콩-B. subtilis D7과 볏짚물-비발아콩을 이용한 경우보다 높게 나타났다. 종균을 이용해 발효된 청국장의 경우 환원당 함량, 아미노태질소 함량, 생균수, protease 활성 및 amylase 활성 등에서 볏짚을 이용한 경우에 비해 더 높게 나타났다. 동물실험을 통한 생산된 청국장의 유효성을 검증한 결과, 모든 청국장은 간세포의 SOD 활성과 혈청 ALP 활성을 증가시켰으며, 발아콩을 이용한 경우에서만 혈청 AST 활성이 증가되었다. 따라서 종균을 이용해 발효된 청국장(특히, 발아콩을 이용한 경우)은 볏짚을 이용해 발효된 청국장에 비해 여러 가지 화학성분이 우수한 것으로 나타나 기능성 성분의 증가와 더불어 품질 향상을 기대할 수 있을 것으로 판단된다. In order to investigate changes in quality and enzyme activity during Chungkookjang fermentation, germinated- and nongerminated yellow soybeans were fermented by Bacillus subtilis and traditional methods. When the soybean was soaked for 6 h and then watered for 4 days with 2 h-interval at 25oC, the highest germination rate was obtained. The germinated soybeans had a higher total isoflavone (988.4 μg/g) than that of the nongerminated soybeans (859.5 μg/g). Amino type nitrogen contents, protease and amylase activities were higher in germinated soybean Chungkookjang, which was fermented with B. subtilis, than nongerminated soybean Chungkookjang, which was fermented with B. subtilis and traditional methods. Reducing sugar and amino type nitrogen contents, the number of viable cells and protease and amylase activities, were higher for Chungkookjang fermented with B. subtilis, than Chungkookjang fermented by traditional methods. ALP and SOD activities in the Chungkookjang diet group were considerably higher than in the control group. AST activity in the germinated soybean Chungkookjang diet group was higher than in the nongerminated soybean Chungkookjang diet group. In conclusion, it is suggested that Chungkookjang prepared with germinated soybeans using B. subtilis D7 could be practically used as a functional product.

콩 종류에 따른 청국장의 발효특성

이나리 ( Na Ri Lee ),이상미 ( Sang Mee Lee ),고태훈 ( Tae Hun Go ),정성윤 ( Seong Yun Jeong ),홍창오 ( Chang Oh Hong ),김근기 ( Keun Ki Kim ),박현철 ( Hyean Cheal Park ),이상몽 ( Sang Mong Lee ),김용균 ( Young Gyun Kim ),손홍주 ( 한국환경과학회 2013 한국환경과학회지 Vol.22 No.6

This study was carried out to investigate changes of protease and amylase activities and nitrogen content in Chungkookjang prepared by Bacillus subtilis S8 and different soybean. Amino-type nitrogen and ammonia-type nitrogen contents increased with an increase in fermentation time and was the highest in black soybean Chungkookjang. The number of viable cells increased up to 24 h of fermentation at all temperatures tested; especially, their levels were the highest at 40℃. Protease activity was the highest in black soybean Chungkookjang. α-amylase activity increased significantly up to 6 h of fermentation at 30℃ and 40℃ and then maintained constantly. It also increased up to 30-36 h of fermentation at 45℃ and then decreased. β -amylase activity was the highest in black soybean Chungkookjang at 35℃and 40℃ and in yellow soybean Chungkookjang at 45℃. Production pattern of reducing sugar was similar to that of β-amylase. Amino-type nitrogen, viable cell number and reducing sugar content and β-amylase activity was the highest in Chungkookjang fermented at 40℃. Considering amino-type and ammonia-type nitrogen contents, Chungkookjang fermentation using yellow soybean was favorable. However, the fermentation using black soybean was favorable, considering protease and amylase activities and reducing sugar content.

Bacillus subtilis D7에 의하여 발효된 백태 청국장의 특성

이나리(Na-Ri Lee),박성보(Sung-Bo Park),이상미(Sang-Mee Lee),고태훈(Tae-Hun Go),황대연(Dae-Youn Hwang),김동섭(Dong-Seob Kim),정성윤(Seong-Yun Jeong),손홍주(Hong-Joo Son) 한국생명과학회 2013 생명과학회지 Vol.23 No.4

본 연구에서는 Bacillus subtilis D7과 발아콩을 이용하여 발효된 청국장의 특성을 조사하였다. 백태는 6시간 침지 후, 25℃에서 2시간마다 4일 동안 물을 공급했을 때, 발아율이 가장 높았다. 총 이소플라본 함량은 발아 전 971.3 μg/g이었고, 발아 후 1023.8 μg/g이었다. 실험에 사용된 모든 청국장에서 아미노태 질소 및 암모니아태질소 함량은 배양시간이 경과함에 따라 증가하였고, 청국장의 pH는 발효과정을 통하여 pH 7.8-8.0로 증가하였다. 청국장의 생균수는 발효 24시간까지 급증한 후, 약간 감소하였다. 청국장의 protease 활성은 발효 30-36시간까지 증가하였고, B. subtilis D7를 이용한 청국장의 α- 및 β-amylase 활성은 발효 12-18시간까지 증가하였다. 청국장을 섭취한 마우스의 ALP 활성과 SOD 활성은 대조군보다 매우 높았다. AST 활성은 미발아콩 청국장 섭취군보다 발아콩 청국장 섭취군에서 더 높았다. 결론적으로 B. subtilis D7을 이용하여 조제된 발아콩 청국장은 품질 특성이 향상되었고, 기능성 성분의 함량이 증가되었다. This study was carried out to investigate the characteristics of white soybean Chungkookjang fermented by Bacillus subtilis D7. The highest germination rate was obtained under 25°C when water was supplied for 4 days at intervals of 2 hr. The total isoflavone content was 971.3 μg/g before germination and 1023.8 μg/g after germination. The amino type- and ammonia type-nitrogen contents of Chungkookjang were proportional to the fermentation time. The pH values of all Chungkookjang soybeans increased up to pH 7.8-8.0 during fermentation. The number of viable cells in all Chungkookjang soybeans increased significantly up to 24 hr. The protease activity of all Chungkookjang soybeans increased up to 30-36 hr. The α- and β-amylase activities of Chungkookjang fermented by B. subtilis D7 were enhanced with increasing fermentation time. Analysis of the effect of Chungkookjang intake on the liver function of mouse showed that the alkaline phosphatase (ALP) activity and the superoxide dismutase (SOD) activity in the Chungkookjang diet group were markedly higher than those in the control group. The asparatate aminotransferase (AST) activity in the germinated soybean Chungkookjang diet group was higher than that in the nongerminated soybean Chungkookjang diet group. Therefore, Chungkookjang soybeans fermented with B. subtilis D7 can be expected to have an increased content of functional components and improved quality characteristics.

수산폐기물 재활용을 위한 미더덕껍질 용매 추출물의 생리활성 탐색

이상미 ( Sang Mee Lee ),강은진 ( Eun Jin Kang ),고태훈 ( Tae Hun Go ),정성윤 ( Seong Yun Jeong ),박근태 ( Geun Tae Park ),이희섭 ( Hee Sup Lee ),황대연 ( Dae Youn Hwang ),정영진 ( Young Jin Jung ),손홍주 ( Hong Joo Son ) 한국환경과학회 2014 한국환경과학회지 Vol.23 No.1

Styela clava tunic is generated in large amounts as a waste from S. clava processing plants and causes environmental problem. Although biological activities of S. clava were reported by many investigators, study on S. clava tunic was little. In this study, therefore, tyrosinase inhibition and antioxidative activities of extracts from S. clava tunic using different solvent were investigated for recycling of the fishery waste. Among extraction methods tested, autoclaved extraction (25.7%) and hot water extraction (18.2%) appeared to be effective for extraction. The highest total phenolic content was 46.6 mg/g in autoclaved extract while the highest flavonoid content was 23.0 mg/g in chloroform extract. All extracts possessed tyrosinase inhibition activity and the inhibition activity was concentration-dependent. Inhibition concentration (IC50) against tyrosinase activity was 0.36×104 mg/ml in ethanol extract, 0.11×103 mg/ml in acetone extract and 0.27 mg/ml in n-butanol extract. Among extracts tested, hot water and autoclaved extracts displayed higher antioxidative activity than organic solvent extracts. Therefore, our data suggest that extract from S. clava tunic may potential candidate for cosmetic product with whitening effect and medicine for diseases caused by various oxidative stresses.

한국전통발효식품인 청국장에서 분리한 Bacillus methylotrophicus에 의한 항산화물질의 생산

이나리 ( Na Ri Lee ),우가영 ( Ga Young Woo ),장준혁 ( Jun Hyeok Jang ),이상미 ( Sang Mee Lee ),고태훈 ( Tae Hun Go ),이희섭 ( Hee Sup Lee ),황대연 ( Dae Youn Hwang ),손홍주 ( Hong Joo Son ) 한국환경과학회 2013 한국환경과학회지 Vol.22 No.7

Although antioxidant activities of Korean traditional fermented foods were reported by many researchers, study on antioxidant activity of microorganism originated from Korean traditional fermented foods was little. Therefore, we improved condition for antioxidant production by a bacterium isolated from home-made Chungkookjang. We selected a bacterial strain, which showed the highest antioxidative activity, from Chungkookjang and then named GJ. The selected GJ strain was identified as Bacillus methylotrophicus by alignment data of 16S rRNA gene nucleotide sequences. Improved medium compositions for DPPH radical scavenging activity were 0.25% sucrose, 1% peptone, 0.01% MgSO4ㆍ7H2O and initial pH 6.5, respectively. Optimal culture conditions were 30℃, 200 rpm and 4% inoculum volume, respectively. In improved conditions, DPPH radical scavenging activity of GJ reached to 91% in a short time. The strain GJ also possessed ACE inhibition and other antioxidative activities; ACE inhibition activity (49.4%), ABTS radical scavenging activity (99.8%), metal chelating activity (67.9%), SOD-like activity (36.5%) and reducing power (A700=5.982) were observed, respectively. Therefore, our results suggest that B. methylotrophicus Gj strain may be potential candidate for functional foods, cosmetic products for anti-aging and medicine for diseases caused by oxidative stress.