Synthesis of the Antihemorrhagic Vitamin by Bacteria

Abstract

Fish meal and rice bran which have been exposed to the action of microörganisms contain the antihemorrhagic vitamin-K. 1 This factor has also been demonstrated in the feces of chicks on a K-free diet. 2 The present work is a preliminary investigation of the production of the vitamin by bacterial synthesis.

It was again found that the putrefaction of ether-extracted, K-free fish meal was accompanied by the formation of appreciable amounts of a fat-soluble, antihemorrhagic substance. Several different types of bacteria were present in the putrefied material. An organism producing a similar putrefaction was isolated in pure culture∗ and inoculated to wet, sterile, ether-extracted fish meal. As a control, a similarly prepared uninoculated fish meal was used. Following 10 days'incubation, when a strongly putrid odor had been developed in the inoculated sample, the material was extracted while still moist by shaking with ether. The ethereal extract, the extracted residue, and the sterile control fish-meal sample were each added to different portions of K-free chick-ration 3 and each portion was fed to day-old chicks for a period of one week. The ethereal extract of putrid fish meal and the extracted residue each contained sufficient antihemorrhagic factor to maintain an approximately normal blood-clotting time of chicks. 3 The sterile fish-meal control gave no evidence of activity.

The fish-meal organism was then inoculated to beef broth, fish-meal broth, proteose-peptone broth, gelatin, and nutrient agar. Washed bacteria from broth and from nutrient agar were found to be rich sources of the antihemorrhagic factor. Liquid media from which the bacteria had ken removed by filtration, and saline used in washing the bacteria were negative. Sterile media used as controls were all negative.