• 미생물학회지
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• 제23권4호
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• pp.297-301
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• 1985

The dependence of activities of Aspergillus phoenicis on the culture conditions in the progesterone transformation reaction was investigated. In the beginning of the reaction, $6{\beta},\;11{\alpha}-dihydroxyprogesterone$ was not produced even at high concentration of $11{\alpha}-hydroxyprogesterone$. However, large amount of the product was obtained after the complete exhaustion of progesterone. When spores of A.phoenicis replaced mycelia as enzyme source, $11{\alpha}-hydroxyprogesterone$ was produced after a considerably long indyction period, and its maximum production rate followed the exponential growth phase. The $6{\beta}-hydroxylation\;of\;11{\alpha}-hydroxyprogesterone$ continued, even after the stationary growth phase. A. phoenicis showed high enzyme activity for these reactions when the phosphate buffer solutions were used in place of the ordinary culture medium. The buffer solutions of low pH gave more yield of $11{\alpha}-hydroxyprogesterone$ than those of high pH. However, the addition of flucose to the buffer solutions did not activate the transformation reaction. The presence of progesterone seems to be necessary for the induction of enzymes for the $6{\beta}-hydroxylation\;of\;11{\alpha}-hydroxyprogesterone\;since\;6{\beta},\;11{\alpha}-dihydroxyprogesterone$ is not produced in the reaction medium containing only $11{\alpha}-hydroxyprogesterone$ as a substrate.

• 한국균학회지
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• 제15권1호
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• pp.23-28
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• 1987

R. nigricans Ehrenberg 에 의한 progesterone 의 $11{\alpha}-hydroxyprogesterone$에 영향을 미치는 여러 물러 화학적 요인에 대하여 조사하였다. Progesterone의 농도가 낮을수록 $11{\alpha}-hydroxyprogesterone$이 최고의 수득율에 도달하는데 소요되는 시간이 짧았으며, 높은 기질 농도에서의 Progesterone은 $11{\alpha}-hydroxyprogesterone$의 hydroxylation을 저해하였다. 이 반응에 대한 최적 pH와 온도 범위는 비교적 넓은 범위로 나타났으며, 균사제의 성장 죄적 조건과 유사하였다. 일정 속도 이상의 진탕 속도와 당류들은 $11{\alpha}-hydroxyprogesterone$ 반응을 촉진시키는 효과를 보였으매, 균사체에서 보다 포자를 효소원으로 사용하였을 때 glucose의 효과는 더 크게 나타났다. 유기 용매는 효소 활성 저해제로 작용하었으며, 저해 정도는 유기 통매 농도가 높을수록, 용매에 처리한 시간이 길어질수록 높게 나타났다.

• 한국균학회지
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• 제13권2호
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• pp.83-87
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• 1985

A. phoenicis는 고온에서도 높은 lactose hydrolysis에 대한 효소 반응 활성을 가지고 있으나, 이 균주에 의한 progesterone의 전환 반응은 반응 온도에 매우 민감한 것으로 나타났다. 균체 내외부의 반응액 조성을 sonication 방법을 이용하여 조사함 결과 반응 초기에 균체의 내부에는 외부에 비하여 $11{\alpha}-hydroxyprogesterone$이 더 높은 농도로 존재하였으나 반응이 진행됨에 따라 균체 내부의 $11{\alpha}-hydroxyprogesterone$이 다른 생성물로 전환되어 오히려 더 낮은 농도의 $11{\alpha}-hydroxyprogesterone$이 존재하였다. A. phoenicis mycelium은 유기용매가 존재하는 반응액내에서도 새로운 액체 배지가 연속적으로 공급될 경우 물질대사를 계속 진행시킬 수 있으며, mycelium의 성장을 저해하지 않는 방법으로 고정된 cell을 이용하여 고정층 반응기에서 연속적으로 A. phoenicis-hydroxyprogesterone을 생산할 수 있다고 판단된다.

• 한국균학회지
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• 제21권1호
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• pp.23-27
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• 1993

Rhizopus nigricans에 의한 progesterone의 $11{\alpha}-hydroxylation$과 ${5\alpha}-reduction$반응에 미치는 이온의 효과를 조사하였다. $Cu^{2+},\;Cd^{2+},\;Co^{2+},\;Mn^{2+},\;Zn^{2+},\;Fe^{2+},\;Mg^{2+},\;Fe^{3+}$ 및 $Na^+$은 $11{\alpha}-hydroxylation$ 반응의 활성을 저하시켰으나 $K^+$은 이 반응을 촉진시켰다. $11{\alpha}-hydroxyprogesterone$을 $5{\alpha}-reduction$시키는 효소는 $Fe^{2+},\;Mn^{2+},\;Mg^{2+},\;Co^{2+},\;Zn^{2+},\;Fe^{2+},\;K^+,\;Na^+$에 의하여 그 활성이 증가되었고 $Cd^{2+}$과 $Cu^{2+}$은 이 반응을 억제하였다. $11{\alpha}-hydroxylation$ 반응을 촉진하는데는 $10^{-3}\;M$의 $K^+$이 효과적이었으며 $5{\alpha}-reduction$반응을 엑제하기 위하여는 $10^{-4}\;M$의 $Cd^{2+}$이 적합하였다. R. nigricans는 세포내의 다효소체계에 의하여 progesterone이 $11\{alpha}-hydroxyprogesterone$으로 전환된 후 $11{\alpha}-hydroxy-allopregnane-3, 20-dione으로 $5{\alpha}-reduction$되므로 $10^{-3}\;M$의 $K^+$을 먼저 첨가하여 2시간 동안 progesterone 전환 반응을 실시한 후에 $10^{-4}\;M$의 $Cd^{2+}$을 첨가하였을 때 가장 높은 $11\{alpha}-hydroxyprogesterone$의 수득률을 얻을 수 있었다.

• 한국균학회지
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• 제11권3호
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• pp.115-119
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• 1983

Aspergillus phoenicis에 의한 progesterone의 전환반응에 대하여 고찰하였다. 본 실험조건에서의 물질대사 결과 $11{\alpha}-hydroxyprogesterone$이 주생성물로 생성 되었다. $11{\alpha}-hydroxyprogesterone$의 농도는 반응시간에 따라 서서히 증가하였으나 40시간 이후부터는 점근값에 도달하였다. 반응액에의 glucose 첨가는 progesterone의 전환반응 속도를 증가시켜 점근값의 농도에 도달하는 시간이 괄목할 만큼 앞당겨졌다. Progesterone의 초기 농도를 증가시키는 것은 전환율이나 반응완결에 소요되는 시간에 큰 변화를 주지 않았으나 전환반응은 progesterone 농도에 대하여 1차반응 형태로 요약될 수는 없었다. 유기용매에 의한 효소 활성의 저하정도는 유기용매의 종류 및 그 농도에 의존하였으며, 낮은 유기용매의 농도에서는 acetone이 가장 높은 전환율을 나타내었다.

• 한국미생물·생명공학회지
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• 제18권1호
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• pp.66-70
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• 1990

부피질호르몬의 제조의 중요한 전구체인 11alpha-hydroxyprogesterone을 Rhizopus nigricans를 이용하여 고농도의 progesterone으로부터 고수율로 생산하기 위한 연구를 수행하였다. 생물전환수율을 최대로 하는 배지조성은 포도당과 casamino acid의 비율이 2:1인 것으로 관찰되었다. Progesterone은 Tween 80 용액에 분산시켜서 종균배양 후 12 시간 정도 지나서 첨가하였을 때 전환수율이 높았다. Progesterone의 농도가 증가함에 따라 생산성이 증가하였고 Progesterone이 20g/$\ell$일 경우 전환수율은 약 70정도 였다.

• 미생물학회지
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• 제29권2호
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• pp.111-116
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• 1991

Rhizopus nigricans produces 11.alpha.-hydroxyprogesternoe with a unidentified byproduct, which is hardly separated. Results of chromatography, IR and NMR spectroscopy identified the byproduct to be 11.alpha.-hydroxy-allopregnane-3,20-dione. R. nigricans was found to transform progesternoe into a monoform intermediate, 11.alpha.-hydroxyprogesterone, from which 11.alpha.-hydroxy-allopregnane-3,20-dione and 6.betha., 11.alpha. - dihydroxyprogesterone were formed respectively by 5.alpha.-reduction and 6.betha.-hydroxylation.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.280-285
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• 1990

Biotransformation of progesterone to 11 $\alpha$ -hydroxyprogesterone by growing cells of Rhizopus nigricans was investigated. As the concentration of progesterone increased, the specific growth rate of R. nigricans decreased linearly, and consequently the conversion yield lowered. The hyphae of the microorganism were observed to become thicker, shorter, and more densely branched at high concentrations of progesterone. In order to improve the process productivity, biotransformation was conducted with continuous addition of progesterone. When the substrate was added continuously at a rate of 0.86 g/hr for 30 hrs, overall conversion yield reached upto 56% while a single addition of the same amount of progesterone yielded about 40% eonversion. When additional feeding of glucose was carried out upon its depletion, an improved br'oconversion yield upto 68% was obtained.

• 미생물학회지
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• 제27권1호
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• pp.70-76
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• 1989

Progestrone bioconversion by immobilized Aspergillus phoenicis was studied. Progesterone was converted into 11$\alpha$-hydroxyprogesterone and 3-minor byproducts. Whole cells of A. phoenicis were immobillized by enreappment with calcium-alginate, K-carrageenan, or polyacrylamide. Of these materials tested, cell immobilized in $Ca^{2+}$ -alginate gels showed the highest activity for 11$\alpha$-hydroxylation of progesterone. In the case of mycelia immobilized in $Ca^{2+}$-alginate, futher progressing hydroxylation of 11$\alpha$-hydroxyprogesterone was greatly reduced. Spores of A. phoenicis which were immobillized with $Ca^{2+}$-alginate and germinatedin situ for 25 hours showed higher 11$\alpha$-hydroxylase activity than those of entrapped whole mycelia and maintained initial enzyme activity for all 8 times of repeated use. After 16 times of reuse, the activity was declined 30% or more. When culture media and $Zn^{2+}$ were introduced into the reaction media, the activity of the immobilized mycelia which had been lowered due to many times of reuse was effectively reactivated.

• 대한유전성대사질환학회지
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• 제14권2호
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• pp.150-155
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• 2014

The measurement of $17{\alpha}$-hydroxyprogesterone ($17{\alpha}$-OHP) in a dried blood spot on filter paper is an important for screening of congenital adrenal hyperplasia (CAH). Since high levels of $17{\alpha}$-OHP are frequently observed in premature infants without congenital adrenal hyperplasia, we evaluated cuts-off based on birth weight and performed validation. Birth weight and $17{\alpha}$-OHP concentration data of 292,204 newborn screening subjects in Greencross labopratories were analyzed. The cut-off values based on birth weight were newly evaluated and validated with the original data. The mean $17{\alpha}$-OHP concentration were 7.25 ng/mL in very low birth weight (VLBW) group, 4.02 ng/mL in low birth weight (LBW) group, 2.53 g/mL in normal birth weight (NBW) group, and 2.24 ng/mL in heavy birth weight (HBW) group. The cut-offs for CAH were decided as follows: 21.12 ng/mL for VLBW and LBW groups and 11.14 ng/mL for NBW and HBW groups. When applied new cut-offs for original data, positive rates in VLBW and LBW groups were decreased and positive rates in NBW and HBW groups were increased. The cut-offs based on birth weight should be used in the screening for CAH. We believe that our new cut-off reduce the false positive rate and false negative rate and our experience for cut-off set up and validation will be helpful for other laboratories doing newborn screening test.