反应详情

Pseudomonas putida D4014-A357 (FERM BP-206) was cultivated in the followingmanner. A medium was prepared by adding 50 ml of tap water to 0.2 g of deoxycholic acid, 0.1 g of glucose, 0.2 g of ammonium nitrate, 0.1 g of potassium dihydrogen phosphate, 0.6 g of dipotassium hydrogen phosphate, 0.02 g of magnesium sulfate heptahydrate and 0.02 g of yeast extract, adjusting the solution to pH 8.4 with 1N sodium hydroxide and further adding water to make the volume 100 ml. This medium was placed in a 500 mlSakaguchi flask and steam-sterilized at 120° C. for 15 minutes. Separately and in advance, the above microbe was grown in the same medium as above on a test tube shaker for one day and a 10 ml portion of the seedculture was added to the above 500 ml Sakaguchi flask, followed by shake culture at 30° C. for 2 days. The precipitate which had formed during the cultivation and the cells were collected by centrifugation and washed with water, and 50 ml of methanol was added thereto to dissolve theprecipitate to a satisfactory extent. Recentrifugation gave a supernatant methanol solution. The methanol was distilled off with a rotary evaporatorto give 65 mg of 12α-hydroxypregna-4-en-3-one-20-carbaldehyde.

1. customA medium was prepared
2. extractionextract
3. additionfurther adding water
4. customfor one day
5. additiona 10 ml portion of the seedculture was added to the above 500 ml Sakaguchi flask
6. waitfollowed by shake culture at 30° C. for 2 days
7. customThe precipitate which had formed during the cultivation
8. customthe cells were collected by centrifugation
9. washwashed with water, and 50 ml of methanol
10. additionwas added
11. dissolutionto dissolve theprecipitate to a satisfactory extent
12. customRecentrifugation gave a supernatant methanol solution
13. distillationThe methanol was distilled off with a rotary evaporatorto