Enzymatic Production of l -Alanine by Pseudomonas dacunhae

Abstract

To establish an advantageous method for the production of l -alanine, a procedure was studied for converting l -aspartic acid to l -alanine by microbial l -aspartic β-decarboxylase. A number of organisms were screened to test their ability to form and accumulate alanine from aspartic acid. Pseudomonas dacunhae was selected as the most advantageous organism. With this organism, enzyme activity as high as 3,910 μliters of CO 2 per hr per ml of medium could be produced by shaking the culture at 30 C in the medium containing ammonium fumarate, sodium fumarate, corn steep liquor, peptone, and inorganic salts. For the enzymatic conversion of l -aspartic acid to l -alanine, the culture broth was employed as the enzyme source. A large amount of l -aspartic acid (as much as 40% of the broth) was converted stoichiometrically to alanine in 72 hr at 37 C. Furthermore, appropriate addition of a surface-active agent to the reaction mixture was found to be highly effective in shortening the time required for the conversion. Accumulated l -alanine was readily isolated in pure form by ordinary procedures with ion-exchange resins. Yields of isolated l -alanine of over 90% from l -aspartic acid were easily attainable.