Sodium Hydroxide → 12alpha-Hydroxypregna-1,4-dien-3-one-20-carbaldehyde

Participants

Reaction Conditions

Reaction Steps

Alcaligenes faecalis D4020-K15 (FERM BP-204) was cultivated in the following manner. A medium (pH 8.4) was prepared by adding tap water to 1.0 g of deoxycholic acid, 0.1 g of glucose, 0.2 g of ammonium nitrate, 0.12 g of potassium dihydrogen phosphate, 0.61 g of dipotassium hydrogen phosphate, 0.02 g of magnesium sulfate heptahydrate, 0.02 g of yeast extract and 0.1 g of sodium hydroxide to a volume of 100 ml. This medium was placed in a 500 ml Sakaguchi 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 seed culture was added to the above-mentioned 500-ml Sakaguchi flask. The flask was shaken at 30° C. for 2 days. The thus-obtained culture broth was centrifuged to separate a mixture of the cells and the precipitate which had formed during the incubation from the culture supernatant. An aqueous 1 N sodium hydroxide solution was added tosaid mixture in an amount sufficient to adjust the pH to 9, and the resultant mixture was extracted with 200 ml of ethyl acetate. On the otherhand, the culture supernatant was adjusted to pH 9 by adding an aqueous 1N sodium hydroxide solution and then extracted with 200 ml of ethyl acetate.Both the extracts were combined and dried over anhydrous sodium sulfate, and the ethyl acetate was distilled off in a rotary evaporator to give 700mg of 12α-hydroxypregna-1,4-dien-3-one-20-carbaldehyde.

Provenance & Evidence