Further selection for a better strain capable of producing D(-)-a-aminobenzylpenicillin (APc) from 6-aminopenicillanic acid (6-APA) was carried out. Pseudomonas melanogenum KY 3987 was consequently selected as a new strain possessing an APc-specific penicillin acylase. The acylase could synthesize APc in good yields from 6-APA and phenylglycine ester and form 6-APA only from APc, not from other common penicillins. Since the Pseudomonas acylase was found incapable of forming penicillin G (Pc-G) from 6-APA and phenylacetic acid, in contrast with E. coli and Kluyvera citrophila enzymes, the enzymatic hydrolysate of Pc-G, for example by K. citrophila cells, which contained 6-APA and phenylacetate, became employed as a source of 6-APA instead of purified 6-APA to synthesize APc by the cells of P. melanogenum.
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Okachi et al. (1973) studied this question.