The present investigation is concerned with the effects of biotin and glucose on glutamic acid fermenation by Microbacterium ammoniaphilum. Both optimal amounts of biotin necessary for maximum growth and maximum accumulation of glutamic acid were determined under the conditoin of various concentration of glucose. As the glucose concentratin was increased, the amounts of biotin required for maximum growth also increased proportionally to the glucose concentrations. The optimal amounts of biotin for maximum accumulation of glutamic acid were smaller than those for maximum growth of cells at any glucose concentration. It is suggested that the process of glutamic acid accumulation is inevitably associated with the process of cell multiplication by both experiments of successive culture of cells grown under the dose of biotin sufficient and deficient for maximum growth.
A newly isolated strain, which belongs to Bacillus, was mutated to adenine dependency by treatment with the mutagen, N-methyl-N'-nitro-N-nitrosoguanidine. The adeninerequiring strains, as expected, accumulated inosine. Cultural conditions for inosine synthesis de novo and the pathway of inosine formation in the mutated strain were investigated. 1) NH4C1 was the preferred nitrogen source. Addition of MnC12.4 H20 was effective for the accumulation of inosine. 2) In this fermentation process, hypoxanthine was formed first, and, after that, inosine was accumulated in the medium. When hypoxanthine was added in the medium, the formation of inosine greatly increased, and, after 90 hours, almost all the hypoxanthine added was converted to inosine. 3) Hypoxanthine-8-C14 was converted to labeled inosine by growing cells, but not converted to cellular RNA. Labeled inosine was formed from hypoxanthine-8-C14 and ribose-1-phosphate by intact cells or cell-free extract. It is presumed that the route of inosine accumulation in the adenine-auxotrophic mutant, Bacillus sp. No. 226, is as follows.
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