EXPRESSION OF RAT-LIVER VITAMIN-D3 25-HYDROXYLASE CDNA IN SACCHAROMYCES-CEREVISIAE

被引：28

作者：

AKIYOSHISHIBATA, M

USUI, E

SAKAKI, T

YABUSAKI, Y

NOSHIRO, M

OKUDA, K

OHKAWA, H

机构：

[1] SUMITOMO CHEM CO LTD,TAKARAZUKA RES CTR,BIOTECHNOL LAB,4-2-1 TAKATSUKASA,TAKARAZUKA,HYOGO 665,JAPAN

[2] HIROSHIMA UNIV,SCH DENT,DEPT BIOCHEM,HIROSHIMA 734,JAPAN

来源：

FEBS LETTERS | 1991年 / 280卷 / 02期

关键词：

VITAMIN-D3; 25-HYDROXYLASE; CYTOCHROME-P450; 5-BETA-CHOLESTANE-3-ALPHA; 7-ALPHA; 12-ALPHA-TRIOL; 27-HYDROXYLASE; HETEROLOGOUS EXPRESSION IN YEAST;

D O I：

10.1016/0014-5793(91)80333-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The cDNA coding for the precursor protein of rat liver mitochondrial vitamin D3 25-hydroxylase, cytochrome P450LMT25, was expressed under the control of the yeast alcohol dehydrogenase 1 promoter and terminator in Saccharomyces cerevisiae AH22 cells. The transformed yeast cells produced a P450LMT25 protein with an almost similar apparent molecular weight as compared with that of the authentic mature enzyme. The expression level of the P450LMT25 hemoprotein was about 5 x 10(4) molecules per cell as determined by reduced CO-difference spectra. The mitochondrial fraction prepared from the transformed yeast cells exhibited both 25-hydroxylase activity toward 1 alpha-hydroxyvitamin D3 and 27-hydroxylase activity toward 5-beta-cholestane-3-alpha, 7-alpha, 12-alpha-triol in a reconstituted system containing bovine adrenodoxin and NADPH-adrenodoxin reductase.

引用

页码：367 / 370

页数：4

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