유가식 배양방법에 의한 재조합 대장균의 고농도 배양을 이용한 Bacillus licheniformis maltogenic amylase ( BLMA ) 의 생산

김명동(Myoung Dong Kim),박관화(Kwan Hwa Park),서진호(Jin Ho Seo) 한국화학공학회 1995 화학공학의이론과응용 Vol.1 No.2

유향 추출물이 MC3T3-E1 세포 활성 및 분화에 미치는 영향

한상헌,김명동,유승한,유용욱,유형근,신형식,Han, Sang-Heon,Kim, Myoung-Dong,You, Seung-Han,You, Yong-Ouk,You, Hyung-Keun,Shin, Hyung-Shik 대한치주과학회 2001 Journal of Periodontal & Implant Science Vol.31 No.2

Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use have been studied for their capacity of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum has the effects to hemostasis, analgesic and anti-inflammatory, and it also has been traditionally used as a drug for the treatment of bone disease in oriental medicine. The purpose of the present study was to investigate the effects of Olibanum extracts on the activity and differentiation of MC3T3-E1 cells, alkaline phosphatase(ALP) synthesis, formation of bone nodules and expression of type I collagen of MC3T3-E1 cells. To examine the cellular activity, MC3T3-E1 cells were cultured with ${\alpha}-MEM(control)$ and each concentration of Olibanum for 2 days and 4 days. To compare the ALP synthesis, MC3T3-E1 cells were cultured with ${\alpha}-MEM(negative\; control)$, dexamethasone(positive control), and each concentration of Olibanum for 2 days and 4 days. To compare the bone nodule formation, MC3T3-E1 ells were cultured for 21 days, and to compare the type I collagen expression, MC3T3-E1 cells were cultured for 4 days. The cellular activity of MC3T3-E1 cells treated with $1{\mu}g/ml$ of Olibanum extracts was significantly increased at 4-day(p<0.05) to control. The activity of ALP in MC3T3-E1 cells treated with $1{\mu}g/ml$ Olibanum extracts was significantly increased at 4-day(p<0.05). All the experimental groups showed much more bone nodule formation than control groups. The group treated with $1{\mu}g/ml$ of Olibanum extracts was the highest bone nodule formation, and showed much more type I collagen expression than negative control. These results indicate that Olibanum extracts may be considered effective in the activity and differentiation of MC3T3-E1 cells.

자궁근종의 치료에 대한 고찰 -2010년 중국내 발표 논문 중심으로-

백영주 ( Young Ju Baek ),김명동 ( Myoung Dong Kim ) 대한한방부인과학회 2011 大韓韓方婦人科學會誌 Vol.24 No.1

수종의 생약추출물이 MC3T3-E1 세포의 염기성 인산분해 효소 활성에 미치는 영향

박상기,김대겸,유승한,김현아,김명동,유형근,신형식,Park, Sang-Kee,Kim, Dae-Kyum,You, Seung-Han,Kim, Hyun-A,Kim, Myoung-Dong,You, Hyung-Keun,Shin, Hyung-Shik 대한치주과학회 2001 Journal of Periodontal & Implant Science Vol.31 No.1

Several growth factors and polypeptides were studied for the regeneration of periodontal supporting tissues which had been lost due to periodontal disease. But these are not commonly used for regenerators of bone tissue or alveolar bone, because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many natural products, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential or periodontal tissues. Cnidii Rhizoma, Rhinocerotis Cornu and Drynariae Rhizoma have been traditionally used as a drug for treatment of bone disease in oriental medicine. The purpose of this study was to examine the ability of alkaline phosphatase synthesis of MC3T3-E1 cells when above medicines were supplimented. MC3T3-E1 cells were cultured with ${\alpha}-MEM(negative control)$, dexamethasone(positive control), and each natural products for 3 and 5 days. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry. Except Cnidii Rhizoma, all of the natural products of this study induced higher activity of ALP synthesis than controls. Among them Drynariae Rhizoma induced the highest activity. In the aspects of culturing time, all medicines did not showed the difference between 3 and 5 days, but $10^{-7}g/ml$ group of Rhinocerotis Corun showed significant increase at 3 days than at 5 days. These results indicate that several natural products have a inducing ability of ALP synthesis on osteoblasts.

김치에서 분리된 젖산균의 β-glucuronidase 활성 탐색

김은정 ( Eun-jung Kim ),신인웅 ( In-ung Shin ),권세영 ( Se-young Kwun ),박은희 ( Eun-hee Park ),이재형 ( Jae-hyoung Yi ),김명동 ( Myoung-dong Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 한국미생물·생명공학회지 Vol.47 No.3

전통발효식품인 김치에서 분리된 젖산균으로부터 β-glucuronidase 효소활성이 높은 균주를 선발하였다. 김치에서 분리된 156점의 젖산균 중 52점의 균주가 glucuronic acid를 탄소원으로 대사하였으며, 대부분의 젖산균은 세포내 β-glucuronidase 활성이 세포외 활성보다 유의적으로 높았다. 순무김치에서 분리된 Leu. mesenteroides KFRI 73007 균주가 0.77 ± 0.01 U/mg protein로서 가장 높은 세포내 β-glucuronidase 효소활성을 나타내었다. 최적 반응조건은 pH 7, 37℃이었으며 1.14 ± 0.01 U/mg protein의 효소활성을 나타냈다. 양이온 금속이온은 β-glucuronidase 효소활성을 약 70% 이상 저해하였으며, 균주 배양에 사용한 탄소원 중 β-glucuronidase 생산을 위한 최적의 탄소원은 glucuronic acid이었다. Lactic acid bacteria (LAB) isolated from kimchi were studied for their β-glucuronidase activity. Among the 156 strains tested, 52 strains utilized glucuronic acid as a carbon source and their intracellular β-glucuronidase activities were significantly higher than their extracellular activities. Leuconostoc mesenteroides KFRI 73007 isolated from turnip kimchi exhibited the highest intracellular β-glucuronidase activity of 0.77 ± 0.01 U/mg protein, which was further increased to 1.14 ± 0.01 U/mg protein under optimized reaction conditions (pH 7, 37℃). The activity of β-glucuronidase was notably decreased by the addition of divalent cations, and glucuronic acid was the best carbon source to produce β-glucuronidase in Leu. mesenteroides KFRI 73007.

막걸리로부터 분리된 Bacillus amyloliquefaciens 균주의 항균 활성

심현수 ( Hyunsu Sim ),김명동 ( Myoung Dong Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2016 한국미생물·생명공학회지 Vol.44 No.1

강원도를 비롯한 전국에서 발효식품 44점으로부터 유해균에 대한 생육억제 활성이 우수한 고초균주를 분리 및 동정 하였으며, 생육억제 활성이 가장 우수하였던 Bacillus amyloliquefaciens MBE1283로 명명된 균주의 유해균에 대한 생육억제 활성, 전분분해 효소활성 및 생육특성을 조사하였다. Escherichia coli O157:H7 KCCM40406을 비롯한 8종류의 유해균에 대한 생육억제 활성을 평가한 결과, 막걸리에서 분리된 B. amyloliquefaciens MBE1283 균주는 다른 B. amyloliquefaciens 균주들에 비해 유의적으로 높은 유해균 생육억제 활성을 나타내었다. B. amyloliquefaciens MBE1283 균주의 α-amylase 효소활성은 대조구 균주들과 비교하였을 때 현저히 낮았으나 glucoamylase 효소활성은 대조구로 사용한 균주와 유사한 수준이었다. B. amyloliquefaciens MBE1283 균주의 최적 생육온도는 45oC로서 대조구 균주보다 온도에 대한 내성이 있는 것으로 판단되었으며, 균체 성장에 적합한 최적 배지는 pH 6이었다. 본 연구를 통하여 막걸리로부터 분리된 B. amyloliquefaciens MBE1283 균주는 여러 종류의 유해균에 항균활성을 지니며 높은 온도와 낮은 pH에 대하여 내성을 지닌 것을 알 수 있었다. The presence of bacterial strains showing antagonistic activity to common pathogens found in a variety of fermented foods in Korea was explored. A bacterium inhibiting the growth of pathogens such as Aspergillus terreus (KCTC6178), A. flavus (KCTC6984), Staphylococcus aureus (KCCM12214), Escherichia coli O157:H7 (KCCM40406), Bacillus cereus (KCTC1012), Cryptococcus neoformans (ATCC208821), Salmonella typhimurium (ATCC19430), and Listeria monocytogenes (KCTC3569) was isolated from Makgeolli, a Korean traditional rice wine. The strain showing high antipathogenic activity was identified as B. amyloliquefaciens based on the nucleotide sequence of the 16S ribosomal RNA gene. Compared with B. amyloliquefaciens KCTC1660, whose genome has been sequenced, the isolate exhibited significantly low activities of starch-degrading enzymes and high resistance to high temperature and low pH.

누룩으로부터 스트레스 내성이 우수한 Pichia farinosa 균주의 분리

권훈주 ( Hun Joo Kwon ),김명동 ( Myoung Dong Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2016 한국미생물·생명공학회지 Vol.44 No.3

누룩으로부터 44℃에서 성장할 수 있는 효모 균주를 분리하고 상대적으로 성장속도가 가장 우수한 MBY/L1569 균주를 선별하였다. MBY/L1569 균주는 Pichia farinosa로 동정 되었으며, 대조구인 P. farinosa KCTC27412 균주에 비하여 내열성, 내산성 및 내알칼리성이 월등히 우수하였다. P. farinosa MBY/L1569 균주는 46℃에서 0.37 ± 0.05 h-1의 비성장속도를 나타내었으며, 배양온도 37℃, 초기 pH 7.0, 포도당 농도 50 g/l 조건에서 0.50 ± 0.02 h-1의 가장 빠른 비성장속도를 나타내었다. 최적 호기적 배양조건에서 P. farinosa MBY/L1569 균주는 50 g/l 포도당으로부터 19.66 ± 0.68 g/l 에탄올을 생산하여 약 40%의 수율을 나타냈다. P. farinosa MBY/L1569 균주는 한국미생물자원센터에 KCTC27753 균주로 기탁하였다. A variety of nuruks collected in different areas in Korea were explored to isolate sixty yeast strains that was able to grow at 44℃. MBY/L1569 strain, which showed the highest growth rate, was selected and identified as Pichia farinosa (Millerozyma farinosa). The isolated strain exhibited superior resistance to heat, acid, and alkali compared with those of P. farinosa KCTC27412 as a control strain. The specific growth rate of P. farinosa MBY/L1569 at 46℃ was 0.37 ± 0.05 h-1, and the highest specific growth rate of 0.50 ± 0.02 h-1 was obtained when it was grown at pH 7.0 and 37℃ with 50 g/l (w/v) glucose as the carbon source. Under optimum growth conditions, strain MBY/L1569 produced ethanol 19.66 ± 0.68 g/l from glucose 50 g/l, with an approximate yield of 40%. P. farinosa MBY/L1569 was deposited at the Korean Collection for Type Cultures as pichia farinosa KCTC27753.

김치로부터 단백질 분해 효소활성이 우수한 Bacillus subtilis 균주의 분리

최찬영 ( Chan-yeong Choi ),김명동 ( Myoung-dong Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 한국미생물·생명공학회지 Vol.45 No.1

전국에서 수집한 발효식품으로부터 단백질 분해 효소활성을 보유한 균주를 분리하였다. Skim milk를 첨가한 TSA 평판배지에서 투명환을 형성한 8점의 균주 중 김치에서 분리된 MBE/L865 균주가 대조구인 KCTC13112 균주에 비하여 약 2.6배의 효소활성을 나타내었으며 Bacillus subtilis로 동정하여 한국미생물보존센터에 KCCM43059 균주로 기탁하였다. B. subtilis KCCM43059 균주의 최적 배양조건은 37℃, pH 8으로 대조구인 KCTC13112 균주의 비성장속도보다 우수하였다. 반응조건 60℃, pH 6에서 429.37 ± 18.65 U/mg protein의 최적 효소활성을 나타내었으며 동일조건에서 대조구 균주보다 효소활성이 우수하였다. 또한 B. subtilisKCCM43059 균주는 대조구 균주에 비하여 NaCl에 상대적으로 높은 내성을 나타내었다. Microbial strains exhibiting proteolytic activity were isolated from kimchi, one of traditional fermented foods in Korea. Eight strains formed clear zones around their colonies when grown on TSA plates supple-mented with skim milk. MBE/L865 exhibited 2.6-fold higher protease activity than that of control strain (Bacillus subtilis KCTC13112). MBE/L865 was identified as B. subtilis and deposited in the Korean Collec-tion for Type Cultures under the accession number of KCCM43059. The optimum growth conditions for B. subtilis KCCM43059 were determined to be 37℃ and pH 8. The strain showed maximum protease activity (429.37 ± 18.65 U/mg protein) at 60℃ and pH 6. Further, B. subtilis KCCM43059 had a higher salt (NaCl) tol-erance than that of the control strain.

김치에서 분리된 젖산균의 β-glucosidase 활성 탐색

장미희 ( Mi Hee Jang ),김명동 ( Myoung Dong Kim ) 한국산업식품공학회 2010 산업 식품공학 Vol.14 No.3

β-Glucosidase 효소활성이 높은 균주를 선발하기 위하여 다양한 김치에서 분리된 젖산균의 β-glucosidase 활성을 탐색하였다. 김치에서 분리된 156개의 젖산균 중 134개의 균주만이 cellobiose를 탄소원으로 대사하였으며, 세포내 β-glucosidase 활성이 세포외 활성보다 현저히 높았다. 배추김치에서 분리된 W. cibaria KFRI88010 균주가 3.7±0.5 unit/mg protein으로서 가장 높은 세포내 β-glucosidase 효소활성을 나타내었으며, 효소활성은 pH 5, 37℃ 반응조건에서 가장 높게 나타났다. Mn2+를 비롯한 금속이온은 효소활성을 크게 저해하였다. W. cibaria KFRI88010 균주를 배양할 때 사용한 탄소원 중, fructose는 cellobiose나 glucose와 비교하여 약 2.5배 이상의 높은 세포내 β-glucosidase 효소활성을 나타내었다. The β-glucosidase (E.C. 3.2.1.21) production capabilities of lactic acid bacteria isolated from a variety of kimchi (fermented vegetables) were examined. When grown in a medium containing cellobiose as carbon source, most lactic acid bacteria showed significantly higher intracellular levels of β-glucosidase than the extracellular levels. A maximum intracellular β-glucosidase activity of 3.7±0.5 (unit/mg protein) was obtained in the case of Weissella cibaria KFRI88010 isolated from kimchi. The optimum reaction conditions for W. cibaria KFRI88010 β-glucosidase activity were pH 5.0 and 37℃, and addition of divalent cations to the reaction mixture resulted in a notable decrease in enzyme activity. The β-glucosidase activity was enhanced twofold when W. cibaria KFRI88010 was grown in a medium containing fructose as compared with to a medium containing glucose or cellobiose.

유가식과 세포재순환 연속공정을 이용한 항혈전제 hirudin의 생산

최치민(Chi-Min Choi),김명동(Myoung-Dong Kim),이상기(Sang-Ki Rhee),서진호(Jin-Ho Seo) 한국생물공학회 1998 KSBB Journal Vol.13 No.4