TY - JOUR
T1 - 1-Ene-steroid reductase of Mycobacterium SP. NRRL B-3805
AU - Goren, T.
AU - Harni, M.
AU - Rimonk, S.
AU - Aharonowitz, Y.
PY - 1983/12
Y1 - 1983/12
N2 - The microbial enzymatic reduction of 1,4-androstadiene-3,17-dione (ADD) to 4-androstene-3, 17-dione (AD), testosterone and 1-dehydrotestosterone (DHT) is described. Two reducing activities observed in washed cell suspensions and cell free extracts of Mycobacterium sp. NRRL B-3805 were found to account for these bioconversions. One was a 1-ene-steroid reductase and the other a 17-keto steroid reductase. The first reducing activity was found to appear in the soluble cell fraction whereas the latter could be precipitated by centrifugation. Maximum 1-ene-steroid reductase specific activity was achieved during the exponential growth phase of the organism and significantly increased upon induction with ADD. The 1-ene-steroid reductase was partially purified (30-fold) by ammonium sulfate fractionation, gel-filtration and ion-exchange chromatography, and was eluted from a Sephacryl S-300 column with an Mr = 115,000. The 1-ene-steroid reductase activity was NADPH-dependent and had specificity towards steroid compounds containing C-1,2 double bond with an apparent Km for ADD of 2.2 × 10-5 M. The reverse reaction catalyzing C-1,2 dehydrogenation could not be detected in our preparations. The results suggest that in Mycobacterium sp NRRL B-3805 and B-3683 the steroid C-1,2 dehydrogenation and 1-ene reduction are two separable activities.
AB - The microbial enzymatic reduction of 1,4-androstadiene-3,17-dione (ADD) to 4-androstene-3, 17-dione (AD), testosterone and 1-dehydrotestosterone (DHT) is described. Two reducing activities observed in washed cell suspensions and cell free extracts of Mycobacterium sp. NRRL B-3805 were found to account for these bioconversions. One was a 1-ene-steroid reductase and the other a 17-keto steroid reductase. The first reducing activity was found to appear in the soluble cell fraction whereas the latter could be precipitated by centrifugation. Maximum 1-ene-steroid reductase specific activity was achieved during the exponential growth phase of the organism and significantly increased upon induction with ADD. The 1-ene-steroid reductase was partially purified (30-fold) by ammonium sulfate fractionation, gel-filtration and ion-exchange chromatography, and was eluted from a Sephacryl S-300 column with an Mr = 115,000. The 1-ene-steroid reductase activity was NADPH-dependent and had specificity towards steroid compounds containing C-1,2 double bond with an apparent Km for ADD of 2.2 × 10-5 M. The reverse reaction catalyzing C-1,2 dehydrogenation could not be detected in our preparations. The results suggest that in Mycobacterium sp NRRL B-3805 and B-3683 the steroid C-1,2 dehydrogenation and 1-ene reduction are two separable activities.
UR - http://www.scopus.com/inward/record.url?scp=0021049111&partnerID=8YFLogxK
U2 - 10.1016/0022-4731(83)90361-8
DO - 10.1016/0022-4731(83)90361-8
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AN - SCOPUS:0021049111
SN - 0022-4731
VL - 19
SP - 1789
EP - 1797
JO - Journal of Steroid Biochemistry
JF - Journal of Steroid Biochemistry
IS - 6
ER -