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Appl Environ Microbiol. 1971 December; 22(6): 992-999
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Enzymatic Production of L-Citrulline by Pseudomonas putida

Department of Applied Biochemistry, Tanabe Seiyaku Co., Ltd., 962, Kashima-cho, Higashiyodogawa-ku, Osaka, Japan
Chemical Research Laboratory, Tanabe Seiyaku Co., Ltd., 962, Kashima-cho, Higashiyodogawa-ku, Osaka, Japan

ABSTRACT

To develop an efficient method for the production of L-citrulline, optimum conditions for the conversion of L-arginine to L-citrulline by microbial L-arginine deiminase and for production of the enzyme were studied. A number of micro-organisms were screened to test their ability to form and accumulate L-citrulline from L-arginine. Pseudomonas putida was selected as the best organism. With this organism, enzyme activity as high as 9.20 units per ml could be produced by a shaking culture at 30 C in a medium containing glucose, ammonium phosphate, L-arginine hydrochloride, yeast extract, peptone, and inorganic salts. Appropriate addition of a surface active agent to the reaction mixture was found to shorten the time required for the conversion. A large amount of L-arginine hydrochloride was converted stoichiometrically to L-citrulline in 62 hr at 37 C. Accumulated L-citrulline was readily isolated in pure form by ordinary procedures with ion-exchange resins. Yields of isolated L-citrulline of over 90.5% from L-arginine hydrochloride were easily attainable.