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Streptomyces griseoplanus SL20209에 의한 Aminopeptidase M 저해제의 생산 조건
고학룡,전효곤,성낙계,고영희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.3
생산용 배지(GSS) 100 ㎖을 포함하는 500㎖ 삼각 플라스크에서 Streptomyces griseoplanus SL20209를 28℃, 3일간 배양했을 때 aminopeptidase M(AP-M) 저해제 생산이 최대에 도달하였으며, 그 이후 pH가 알카리성으로 변하면서 다소 감소하였다. Arabinose, xylose, mannose 및 soluble starch가 저해제 생산에 좋은 탄소원인 반면, glucose는 균생육에는 좋으나 저해제 생산은 강하게 저해하였다. Soybean meal, fish meal, gluten meal 및 peanut powder와 같은 천연 유기질소원이 저해제 생산에 좋은 질소원이었으며, soytone이나 peptone 및 NH_4Cl과 NH_4NO_3와 같은 무기질소원이 함유된 배지에서는 저해제 생산이 빈약하였다. 배지내에 각각 0.5%(w/v)의 yeast extract난 0.05%의 K_2HPO_4의 첨가는 균생육을 촉진시켜 저해제 생산을 증가시켰으며, CaCo_3(0.3%)와 zeolite(0.5%)도 저해제 생산 증가의 효과를 나타내었다. 저해제 생산을 위한 최적 배양온도 및 초기 pH 범위는 각각 28℃와 6.0~7.0이었다. Arabinose(ASY 배지) 또는 soluble starch(SSY 배지) 3%, SBM 2.5%, yeast extract 0.5%, K_2HPO_4 0.05%, CaCO_3 0.1% 및 zeolite 0.3%(pH 6.8)의 조성의 두 개선된 배지에서 초기의 GSS 배지에서 보다 저해제 생산이 1.8배 증가되었다. Maximum amount of the aminopeptidase M inhibitors produced by Streptomyces griseoplanus SL20209 in 500 ml-Erlenmeyer flask was accumulated after cultivation for 3 days at 28℃, thereafter the amount of inhibitors decreased slowly with a pH change to alkaline. Arabinose, xylose, mannose and soluble starch were good carbon sources for the production of the inhibitors. On the other hand, glucose was only good for the cell growth but potently inhibited the production of inhibitors. Natural organic nitrogen sources such as soybean meal, fish, meal, gluten meal and peanut powder were good for the production of inhibitors, however, soytone, peptone and inorganic nitrogens such as NH_4Cl and NH_4NO_3 were poor. Inclusion of yeast extract (0.5%, w/v) or K_2HPO_4 (0.05%) into the production medium increased the production of inhibitors by accelerating cell growth. The production of inhibitors was slightly increased on the medium containing CaCo_3(0.3%) and zeolite (0.5%), respectively. Optimal temperature and initial pH range for the production of inhibitors were determined to be 28℃ and 6.0~7.0, respectively. Employing two improved production media consisting of 3% arabinose or soluble starch, 2.5% soybean meal, 0.5% yeast extract, 0.05% K_HPO_4 0.1% CaCO_3 and 0.3% zeolite (pH 6.8), 1,8-fold increase in the amount of inhibitors was achieved, comparing with the basal medium used.
Aminopeptidase M 저해제인 Valistatin과 des-Asp^4-Amastatin을 생산하는 방선균 SL20209의 특성 및 동정
고학룡,전효곤,정명철,서현효,김홍중,박용하,고영희 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1
Aminopeptidase M 저해제인 valistatin과 des-asp^4-amastatin을 생산하는 방선균 분리주 SL20209의 특성을 조사하고 수리동정을 실시하였다. 형태적, 배양학적 및 생리적 특성으로 부터 SL20209는 방성균 중에서도 Streptomyces 속에 속하는 것으로 나타났으며, 43개의 분류단위 특성으로 TAXON program을 이용하여 수리동정한 결과, 주군집 29의 Streptomyces 중 Streptomyces griseoplanus와 가장 근접한 것으로 나타났다. 따라서, 분리주 SL20209를 S. griseoplanus SL20209로 동정하였다. Characterization and numerical identification were carried out for an actionmycetes SL20209. Morphological, cultural and physiological properties of SL20209 which produced valistatin and des-asp^4-amastatin as inhibitors of aminopeptidase M were evaluated. The isolate was identified to be the genus of Streptomyces. Fourty-three taxonomic units were analysed by using a TAXON program. The isolate was classified into the major cluster 29 of Streptomyces and best-matched to Streptomyces griseoplanus.
DNA Rearrangement of TOL Plasmid in Pseudomonas putida PpG1 Harbouring CAM Plasmid
Chun, Hyo Kon,Cho, Kyung Yun,Kho, Yung Hee 한국산업미생물학회 1990 한국미생물·생명공학회지 Vol.18 No.4
접합에 의해 P. putida mt-2의 TOL를 CAM 함유 P. putida PpG1으로 이동시켜 형성된 접합주 P. putida CST3A는 작아진 TOL(TOL△)를 가지고 있었지만 m-toluate를 분해할 수 있었다. 접합에 의한 이동실험과 불화합성 시험결과, TOL의 m-toluate 분해유전자는 CAM에 결합되어 CAM : : TOL^* 플라스미드를 형성하고 있었다. 불화합성 Inc P9 군에 속하는 NAH를 CAM: : TOL^*과 TOL△을 가지는 P. putida CST3A로 이동시키면 TOL△의 방출이 관찰되었으나 m-toluate 대사에는 아무런 영향을 미치지 않았다. The TOL plasmid, pWWO, conjugally transferred from Pseudomonas putida mt-2 was dissociated into TOL^* and TOL △ in P. putida PpG1 carrying CAM plasmid. The TOL^* was integrated into the CAM plasmid, and the resulting plasmid was designated as CAM::TOL^*. The introduction of NAH plasmid, belonging to Inc P9 incompatibility group, into P. putida CST3A carrying CAM::TOL^* plasmid and TOL △ plasmid did not affect m-toluate catabolism, but resulted in expelling the TOL △ plasmid.
Occurrence of OF4949II in the Fungal Mat formed by Surface Culture of Aspergillus niger F-580
CHUN, HYO KON,CHUNG, MYUNG CHUL,KO, HACK RYONGKo, Hack Ryong,LEE, HO JAELee, Ho Jae,KHO, YUNG HEEKho, Yung Hee 한국미생물 · 생명공학회 1995 Journal of microbiology and biotechnology Vol.5 No.5
Aspergillus niger F-580, a potent producer of aminopeptidase M inhibitor, was isolated from the brown spots of plant leaves with a pathological trait. The inhibitory activity was found only in the fungal mat formed by surface culture of Aspergillus niger F-580, but not in the culture supernatant or cell pellet. The inhibitor was purified from the hot water extract of this fungal mat by using chromatographies on Diaion HP-20, DEAE-cellulose, Sephadex G-10 and YMC-ODS-AQ columns. The purified inhibitor was analyzed by UV, mass, and NMR spectroscopies, and identified as OF494911, which had been isolated as an aminopeptidase B inhibitor from Penicillium rugulosum OF4949
Chun, Hyo Kon,Ko, Hack Ryong,Moon, Hang Sick,Kho, Yung Hee 한국미생물 · 생명공학회 1995 Journal of microbiology and biotechnology Vol.5 No.6
Two types of Rhizoctonia solani esterases induced by cutin hydrolysate were partially purified by ammonium sulfate precipitation and gel filtration. The esterase Ⅰ with hydrolyzing activity toward both ρ-nitrophenyl butyrate and ρ-nitrophenyl palmitate and the esterase Ⅱ with hydrolyzing activity toward only ρ-nitrophenyl butyrate were inhibited by ebelactone B, an esterase inhibitor produced by actinomycetes with IC_50 values of 0.01 and 0.09 ㎍/㎖, respectively. Spraying on rice seedling with ebelactone B at a concentration of 30 ㎍/㎖ completely suppressed infection by R. solani. Ebelactone B could not protect the wounded rice seedling and did not show any inhibitory effect on the mycelial growth at a concentration of 1 ㎍/㎖. These results indicate that ebelactone B, an esterase inhibitor protects rice plants from infection with R. solani by inhibition of penetration, not through fungitoxic or fungicidal effect.
Chun, Hyo Kon,Cho, Kyung Yun,Kho, Yung Hee 한국미생물 · 생명공학회 1994 Journal of microbiology and biotechnology Vol.4 No.4
Pseudomonas putida 3SK, which was constructed by the conjugal transfer of the CAM::TOL^* plasmid of Pseudomonas putida CST3A and the NAH plasmid of Pseudomonas putida KCTC 2403 into n-alkane assimilating Pseudomonas putida KCTC 2405, showed a broad degradation spectrum and floc-forming ability. This strain degraded m-toluic acid, naphthalene, camphor and decane simultaneously. Hg^2+ at the concentration of 1 ppm in the minimal medium could not inhibit the growth of this strain. The degradation of m-toluic acid by Pseudomonas putida 3SK was not repressed by the easily utilizable compounds, such as glucose and succinate. But, the addition of formalin inhibited the growth of Pseudomonas putida 3SK. After the cultivation of this strain on the artificial wastewater containing m-toluic acid, naphthalene, camphor and decane for 24 hr, the initial COD value (1500) of the artificial wastewater was declined to 300.
The Complete Nucleotide Sequence of Alkalophilic Bacillus sp. K-17 β-Xylosidase Gene
Chun, Hyo Kon,Ko, Hak Ryong,Kho, Yung Hee,Sung, Nack Kie 한국미생물 · 생명공학회 1991 Journal of microbiology and biotechnology Vol.1 No.1
The complete nucleotide sequence of alkalophilic Bacillus sp. K-17 β-xylosidase gene and its flanking regions were established. A 1263-bp of an open reading frame for β-xylosidase was observed. The molecular weight (50, 521 dalton), deduced from the nucleotide sequence of β-xylosidase gene, agreed with the result obtained by SDS-polyacrylamide gel electrophoresis of the purified enzyme (51,000 dalton). The Shine-Dalgarno sequence, 5'-GAGGAGG-3', was found 8 bp upstream of the initiation codon ATG. The -10 sequence (TAAAAT) in the promoter region for β-xylosidase gene was similar to the consensus sequence for Bacillus subtilis RNA polymerase, whereas the -35 sequence (TCGATCA) different from all the known -35 regions in the promoter for Bacillus subtilis RNA polymerase.
Tungtungmadic Acid, a Novel Antioxidant, from Salicornia herbacea
정영철,Hyo Kon Chun,Jae Young Yang,Ji Young Kim,한은희,Yung Hee Kho,Hye Gwang Jeong 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.10
Tungtungmadic acid (3-caffeoyl-4-dihydrocaffeoyl quinic acid) is a new chlorogenic acid derivative that was isolated from the Salicornia herbacea. The structure of tungtungmadic acid was determined using chemical and spectral analysis. The antioxidant activity of tungtungmadic acid was evaluated using various antioxidant assays, including free radical scavenging, lipid peroxidation and hydroxyl radical-induced DNA strand breaks assays. Tungtungmadic acid (IC50 = 5.1 µM and 9.3 µM) was found to have higher antioxidant activity in the DPPH scavenging assay as well as in the iron-induced liver microsomal lipid peroxidation system. In addition, the tungtungmadic acid was also effective in protecting the plasmid DNA against strand breakage induced by hydroxyl radicals.