Enzymatic production of L-citrulline by Pseudomonas putida.
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.
D000388:Air; D001120:Arginine / Q000378:metabolism; D001419:Bacteria / Q000378:metabolism; D001965:Bromides; D002854:Chromatography, Paper; D002956:Citrulline / Q000096:biosynthesis*; D003470:Culture Media; D004790:Enzyme Induction; D005285:Fermentation; D005658:Fungi / Q000378:metabolism; D005947:Glucose; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000378:metabolism*; D010461:Peptones; D010710:Phosphates; D011549:Pseudomonas / Q000201:enzymology / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds; D012440:Saccharomyces; D013045:Species Specificity; D013237:Stereoisomerism; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
D001965:Bromides; D003470:Culture Media; D010461:Peptones; D010710:Phosphates; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; D002956:Citrulline; D001120:Arginine; D006867:Hydrolases; D005947:Glucose