To establish a novel process for the production of L-glutamic acid from n-paraffins, a glycerol auxotroph GL-21, a new type mutant, was successfully obtained from Corynebacterium alkanolyticum No. 314 by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. This auxotroph required glycerol for its growth regardless of the carbon source used. At 72 hr, this mutant GL-21 produced about 40mg/ml of L-glutamic acid from n-paraffins in the culture broth at 0.01 per cent addition of glycerol in the absence of penicillin. A thiamine auxotroph, a biotin auxotroph and an oleic acid auxotroph were also obtained by a similar technique, but these auxotrophs were found to be inapplicable for theproduction of L-glutamic acid from n-paraffins.