Efficient enzymatic synthesis of ampicillin using mutant Penicillin G acylase with bio-based solvent glycerol

•Bio-based solvent glycerol was used for ampicillin synthesis.•An ampicillin yield of 93.5% was achieved at low acyl donor/nucleophile ratio (1.05:1).•1.12 mol acyl donor D-PGME was required to produce 1 mol ampicillin.

To fulfill the industry demand of ampicillin enzymatic synthesis, immobilized mutant Penicillin G acylase and bio-based solvent glycerol were employed at high substrate concentration and low acyl donor/nucleophile ratio. After process optimization, good yield and low operation costs were achieved.

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