핵산 및 그 관련물질 연구의 일환으로서 미생물 효소에 의하여 핵산을 분해하여 5'-mononucleotides 생성을 목적으로 미생물에서 5'-phosphodiesterase 생성균주를 얻기위하여 전국 각지 76개 지역에서 논, 밭, 산, 하천의 토양 그리고 퇴비, 누룩, 메주 등 시료 210종을 수집하였다. 이 수집 시료로 부터 dilution pour plate method 로서 Aspergillus속 240주, Penicillium속 232주, Neurospora속, 19주, Monascus속 16주, 그리고 Streptomyces속 265주로 총 758주를 순수 분리하였다. 분리된 모든 균주에 대하여 RNA-depolymerase 균의 crude enzyme 중에는 5'-AMP deaminase가 공존하고 있으므로 효소반응 중에 RNA 가 5'-mononucleotide로 분해 축적하는 동안에 5'-AMP의 adenine의 6위치에 붙은 $NH_2$기를 탈아미노하여 hypoxanthine으로 하기 때문에 5'-IMP 로 되는 것이라 생각된다. 분리 동정 된 Penicillium citreo-viride PO 2-11 strain과 Streptomyces aureus SOA 4-21 strain 이 생성한 5'-phosphodiesterase 로서 RNA를 효소 분해하여 5'-mononucleotide 의 정량한 결과는 Table 10과 같다. productivities를 1차 screening 하고 5'-phosphodiesterase productivities로서 2차 screening 하여 우수균주를 얻고 동정하였다. 우수균주의 5'-phosphodiesterase productivity에 대하여 배양상의 optimum condition을 검토하고 5'-phosphodiesterse activity에 미치는 여러 화합물의 영향과 효소반응의 최적 조건을 구명하였다. 우수균주가 생성한 5'-phosphodiesterase에 의하여 RNA 분해로 반응 최종 산물을 ion exchange column chromatography법으로 정량하고 최종 분해 산물엔 5'-mononucleotide를 paper chromatography, thinlayer chromatography, UV-absorption spectra, carbazole reaction 및 Schiff's reaction 등으로 동정한 결과는 다음과 같다. [1] 5'-phosphodiesterase productivity가 가장 우수한 두 균주를 선정하였고 이들은 토양에서 분리 되었으며 선정된 푸른 곰팡이는 Penicillium citreoviride PO 2-11로 동정되었고 방사선균은 Streptomyces aureus SOA 4-21로 동정되었다. [2] 분리 선정 된 Penicillium citreo-viride PO 2-11 strain은 배양상의 optimum condition 이 pH 5.0이고 temperature는 $30^{\circ}C$이었고 이 균이 생성한 5'-phosphodiesterase의 효소 반응상의 optimum condition 이 pH 4.2이고 temperature는 $60^{\circ}C$이었다. 그리고 5'-phosphodiesterase 생성에서 최적 탄소원은 sucrose이고 질소원은 $NH_4NO_3$이고 corn steep liquor나 혹은 yeast extract를 각각 0.01%씩 첨가한 구는 첨가하지 않은 control 구보다 20%의 5'-phosphodiesterase 생성 증가를 나타 내었다. 이 균이 생성한 5'-phosphodiestrase는 $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ 등 금속이온은 activator이고 EDTA, citrate, $Cu^{++},\;Co^{++}$ 등은 inhibitor 임을 알았다. 이 균이 생성한 5'-phosphodiesterase는 RNA를 분해하여 분해율 65.81%이었고 5'-AMP, 5'-GMP, 5'-UMP 및 5'-CMP를 생성하며 이때 축적되는 5'-mononucleotides중 5'-GMP 만이 정미성이 있음을 알았고 이균은 5'-AMP deamaminase가 없음을 확인하였다. 이 균의 효소에 의하여 RNA에서 정미성 5'-GMP 186.7 mg/RNA(g)를 생산할수 있음을 알았다. [3] 분리 선정된 Streptomyces aureus SOA 4-21 strain은 배양상의 optimum condition이 pH 7.0이고 temperature는 $28^{\circ}C$이었고 이 균이 생성한 5'-phosphodiesterase의 효소반응상의 optimum condition이 pH 7.3이고 temperature는 $50^{\circ}C$이었다. 그리고 5'-phosphodiesterase 생성에서 최적탄소원은 glucose이고 질소원은 asparagine이고 yeast extract 0.01%첨가구가 control 구보다 40%의 5'-phosphodiesterase 생성증가를 나타내었다. 이 균이 생성한 5'-phosphodiesterase는 $Ca^{++},\;Zn^{+
핵산 및 그 관련물질 연구의 일환으로서 미생물 효소에 의하여 핵산을 분해하여 5'-mononucleotides 생성을 목적으로 미생물에서 5'-phosphodiesterase 생성균주를 얻기위하여 전국 각지 76개 지역에서 논, 밭, 산, 하천의 토양 그리고 퇴비, 누룩, 메주 등 시료 210종을 수집하였다. 이 수집 시료로 부터 dilution pour plate method 로서 Aspergillus속 240주, Penicillium속 232주, Neurospora속, 19주, Monascus속 16주, 그리고 Streptomyces속 265주로 총 758주를 순수 분리하였다. 분리된 모든 균주에 대하여 RNA-depolymerase 균의 crude enzyme 중에는 5'-AMP deaminase가 공존하고 있으므로 효소반응 중에 RNA 가 5'-mononucleotide로 분해 축적하는 동안에 5'-AMP의 adenine의 6위치에 붙은 $NH_2$기를 탈아미노하여 hypoxanthine으로 하기 때문에 5'-IMP 로 되는 것이라 생각된다. 분리 동정 된 Penicillium citreo-viride PO 2-11 strain과 Streptomyces aureus SOA 4-21 strain 이 생성한 5'-phosphodiesterase 로서 RNA를 효소 분해하여 5'-mononucleotide 의 정량한 결과는 Table 10과 같다. productivities를 1차 screening 하고 5'-phosphodiesterase productivities로서 2차 screening 하여 우수균주를 얻고 동정하였다. 우수균주의 5'-phosphodiesterase productivity에 대하여 배양상의 optimum condition을 검토하고 5'-phosphodiesterse activity에 미치는 여러 화합물의 영향과 효소반응의 최적 조건을 구명하였다. 우수균주가 생성한 5'-phosphodiesterase에 의하여 RNA 분해로 반응 최종 산물을 ion exchange column chromatography법으로 정량하고 최종 분해 산물엔 5'-mononucleotide를 paper chromatography, thinlayer chromatography, UV-absorption spectra, carbazole reaction 및 Schiff's reaction 등으로 동정한 결과는 다음과 같다. [1] 5'-phosphodiesterase productivity가 가장 우수한 두 균주를 선정하였고 이들은 토양에서 분리 되었으며 선정된 푸른 곰팡이는 Penicillium citreoviride PO 2-11로 동정되었고 방사선균은 Streptomyces aureus SOA 4-21로 동정되었다. [2] 분리 선정 된 Penicillium citreo-viride PO 2-11 strain은 배양상의 optimum condition 이 pH 5.0이고 temperature는 $30^{\circ}C$이었고 이 균이 생성한 5'-phosphodiesterase의 효소 반응상의 optimum condition 이 pH 4.2이고 temperature는 $60^{\circ}C$이었다. 그리고 5'-phosphodiesterase 생성에서 최적 탄소원은 sucrose이고 질소원은 $NH_4NO_3$이고 corn steep liquor나 혹은 yeast extract를 각각 0.01%씩 첨가한 구는 첨가하지 않은 control 구보다 20%의 5'-phosphodiesterase 생성 증가를 나타 내었다. 이 균이 생성한 5'-phosphodiestrase는 $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ 등 금속이온은 activator이고 EDTA, citrate, $Cu^{++},\;Co^{++}$ 등은 inhibitor 임을 알았다. 이 균이 생성한 5'-phosphodiesterase는 RNA를 분해하여 분해율 65.81%이었고 5'-AMP, 5'-GMP, 5'-UMP 및 5'-CMP를 생성하며 이때 축적되는 5'-mononucleotides중 5'-GMP 만이 정미성이 있음을 알았고 이균은 5'-AMP deamaminase가 없음을 확인하였다. 이 균의 효소에 의하여 RNA에서 정미성 5'-GMP 186.7 mg/RNA(g)를 생산할수 있음을 알았다. [3] 분리 선정된 Streptomyces aureus SOA 4-21 strain은 배양상의 optimum condition이 pH 7.0이고 temperature는 $28^{\circ}C$이었고 이 균이 생성한 5'-phosphodiesterase의 효소반응상의 optimum condition이 pH 7.3이고 temperature는 $50^{\circ}C$이었다. 그리고 5'-phosphodiesterase 생성에서 최적탄소원은 glucose이고 질소원은 asparagine이고 yeast extract 0.01%첨가구가 control 구보다 40%의 5'-phosphodiesterase 생성증가를 나타내었다. 이 균이 생성한 5'-phosphodiesterase는 $Ca^{++},\;Zn^{+
As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of di...
As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.
As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.
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