YAKHAK HOEJI (약학회지)

The Pharmaceutical Society of Korea (대한약학회)
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Isolation and Partial Purification of the Steroid 9${\alpha}$-Hydroxylase from Mycobacterium fortuitum

Mycobacterium fortuitum의 스테로이드 9${\alpha}$-하이드록실라제의 분리 및 부분정제

  • Published : 1997.10.01
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Abstract

The steroid 9${alpha}$-hydroxylase activity has been detected in cytosol fraction, $100,00{\times}g$ supernatant of cell free extract of Mycobacterium fortuitum. The activity was not linear with protein concentration in the assay suggesting 9${alpha}$-hydroxylase is a multicomponent enzyme. The 9${alpha}$-hydroxylase system was partially purified through fractional saturation of ammonium sulfate, strong anion exchange (Mono Q) column chromatography, gel filtration (Superose 12) column chromatography, and testosterone affinity gel chromatography. Ammonium sulfate 50~60% saturated fraction of the cytosol gave 9${alpha}$-hydroxylase activity. For further purification, the half-saturated ammonium sulfate fraction was applied to Mono Q, Superose 12, or affinity gel column. The purification factors of 9${alpha}$-hydroxylase containing fraction after Mono Q, Superose 12, and affinity gel chromatography was 13, 11, and 17 respectively.

Keywords

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