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Purification of the NADH reductase component of the steroid 9α-hydroxylase fromMycobacterium fortuitum

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Abstract

The NADH reductase component of the steroid 9α-hydroxylase fromMycobacterium fortuitum was purified to homogeneity. Recovery of the enzyme from the 50≈60% ammonium sulfate saturated fraction was 49%, with a purification factor of 100-fold. The NADH reductase has a relative molecular mass of 60 KDa as determined by SDS-PAGE. The absorption maxima at 410 and 450 nm indicate the presence of iron-sulfur group and flavin. These prosthetic groups seemed to function as redox groups that transfer electrons from NADH to the following protein. The KM value for NADH as substrate was 68 μM. The NH2-terminal amino acid sequence of the reductase was determined as Met-Asp-Ala-Ile-Thr-Asn-Val-Pro-Leu-Pro-Ala-Asn-Glu-Pro-Val-His-Asp-Tyr-Ala-Thr. This sequence does not show a homology with the NH2-terminal sequences reported for the reductase component of other monooxygenases, suggesting that the NADH reductase component of the steroid 9α-hydroxylase system is novel.

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  1. College of Pharmacy, Seoul National University, 151-742, Seoul, Korea

    Hee Kyoung Kang & Sang Sup Lee

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  1. Hee Kyoung Kang
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  2. Sang Sup Lee
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Kang, H.K., Lee, S.S. Purification of the NADH reductase component of the steroid 9α-hydroxylase fromMycobacterium fortuitum . Arch. Pharm. Res. 20, 590–596 (1997). https://doi.org/10.1007/BF02975217

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