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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.
Kho, Yung-Hee,Chung, Myung-Chul,Chun, Hyo-Kon,Lee, Choong-Hwan,Lee, Ho-Jae,Kim, Su-Il The Korean Society for Microbiology and Biotechnol 1995 Journal of microbiology and biotechnology Vol.5 No.3
The effect of carbon and nitrogen sources on the production of novel aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 has been studied. High D-glucose and ammonia concentrations (5$\%$/> and 1$\%$/>, respectively) exerted a negative influence on the inhibitor formation. The suppressive effect of glucose on the inhibitor formation is probably caused by an effect of medium pH rather than that of cyclic AMP. To establish the optimum conditions for inhibitor overproduction, various nitrogen sources and ammonium ion-trapping agents were examined. The use of ammonia slow-releasing nitrogen sources such as soybean meal and fish meal, or ammonium ion-trapping agents such as kaoline, celite, and natural zeolite achieved the enhancement of inhibitor production. These results also indicate that inhibitor formation is affected by ammonium ion repression.