Sasaki J, Miyazaki A, Saito M, Adachi T, Mizoue K, Hanada K, Omura S
Research Center, Taisho Pharmaceutical Co., Ltd., Saitama, Japan.
Appl Microbiol Biotechnol. 1992 Nov;38(2):152-7. doi: 10.1007/BF00174460.
To enzymatically synthesize active metabolites of vitamin D3, we screened about 500 bacterial strains and 450 fungal strains, of which 12 strains were able to convert vitamin D3 to 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3] via 25-hydroxyvitamin D3 [25(OH)D3]. The conversion activity was only detected in strains belonging to the genus Amycolata among all the organisms tested. A preparative-scale conversion of vitamin D3 to 25(OH)D3 and 1 alpha,25(OH)2D3 in a 200-1 tank fermentor using A. autotrophica FERM BP-1573 was accomplished, yielding 8.3 mg 25(OH)D3/l culture and 0.17 mg 1 alpha,25(OH)2D3/l culture. A related compound, vitamin D2, could be also converted to 25-hydroxyvitamin D2 and 1 alpha,25-dihydroxyvitamin D2 using the same strain. The cytochrome P-450 of FERM BP-1573 was detected by reduced CO difference spectra in whole-cell suspensions. Vitamin D3 in the culture induced cytochrome P-450 and the conversion activity simultaneously, suggesting that the hydroxylation at C-25 of vitamin D3 and at C-1 of 25(OH)D3 originates from cytochrome P-450.
为了酶促合成维生素D3的活性代谢产物，我们筛选了约500株细菌菌株和450株真菌菌株，其中有12株能够通过25-羟基维生素D3 [25(OH)D3]将维生素D3转化为1α,25-二羟基维生素D3 [1α,25(OH)2D3]。在所有测试的生物体中，仅在属于无枝菌酸菌属的菌株中检测到转化活性。使用自养无枝菌酸菌FERM BP-1573在200升发酵罐中进行了维生素D3到25(OH)D3和1α,25(OH)2D3的制备规模转化，每升培养物产生8.3毫克25(OH)D3和0.17毫克1α,25(OH)2D3。使用同一菌株，相关化合物维生素D2也可转化为25-羟基维生素D2和1α,25-二羟基维生素D2。通过全细胞悬浮液中的还原型CO差光谱检测到FERM BP-1573的细胞色素P-450。培养物中的维生素D3同时诱导细胞色素P-450和转化活性，这表明维生素D3在C-25位和25(OH)D3在C-1位的羟基化源自细胞色素P-450。
