Factors affecting the synthesis of ampicillin and hydroxypenicillins by the cell-bound penicillin acylase of Escherichia coli.
1. The effect of pH, temperature, reactant concentration and reaction time has been investigated for the synthesis of benzylpenicillin, dl-alpha-hydroxybenzylpenicillin and d-alpha-aminobenzylpenicillin from 6-aminopenicillanic acid by the penicillin acylase of Escherichia coli. 2. Synthesis of penicillins from carboxylic acids proceeds most rapidly at pH5; with amides the optimum pH is higher (6-7) but the reverse reaction rapidly sets in. This can be counteracted by lowering the pH or adding more amide. Optimum temperatures are 35-40 degrees . 3. Most rapid synthesis of penicillin was obtained with the N-acylglycine and methyl ester derivatives of carboxylic acids. Increasing the amide/6-APA ratio above 1:1 raised the rate of synthesis of penicillins. 4. Preferential synthesis of d-alpha-hydroxybenzylpenicillin takes place in a reaction mixture containing dl-mandelic acid. 5. From d- and l-mandelamide, d- and l-alpha-hydroxybenzylpenicillins were prepared, the former being more bioactive than the latter. p-Hydroxy- and 3,4-dihydroxybenzylpenicillins were also prepared, the latter being more active against some Gram-negative bacteria than benzylpenicillin.
D000577:Amides / Q000378:metabolism; D000581:Amidohydrolases / Q000378:metabolism*; D000667:Ampicillin / Q000096:biosynthesis*; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008333:Mandelic Acids; D010406:Penicillins / Q000096:biosynthesis*; D013696:Temperature
D000577:Amides; D008333:Mandelic Acids; D010406:Penicillins; D000667:Ampicillin; D000581:Amidohydrolases