Acetyl esterase from mediterranean oranges: partial purification,immobilisation and biotransformations |
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Authors: | Piero Pasta Roberto Verga Francesca Zambianchi Moreno Daminati |
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Institution: | 1. Istituto di Chimica del Riconoscimento Molecolare CNR, Via Mario Bianco 9, 20131, Milano, Italypiero.pasta@icrm.cnr.it;3. Antibioticos S.p.A., Strada Rivoltana km 6/7, 20090, Rodano (Milano), Italy;4. Istituto di Chimica del Riconoscimento Molecolare CNR, Via Mario Bianco 9, 20131, Milano, Italy;5. Resindion S.r.l., Via Roma 55, 20082, Binasco (Milano), Italy |
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Abstract: | Acetyl esterase (acetic-ester acetylhydrolase, EC 3.1.1.6) from citrus peel, whose natural role is not well known, catalyses, in vitro, the hydrolysis of acetyl groups from a wide range of substrates. This enzyme was extracted from Mediterranean orange peel, largely available in Italy, and purified 190-fold by a single chromatographic step on Sepabeads FP-HG. SDS polyacrylamide gel electrophoresis of the purified enzyme showed a major protein band, corresponding to a molecular mass of 45 kDa. Both free and immobilised enzyme were used in biotransformations. The enzyme removed the acetyl group in the 3 position of β-lactamic antibiotics, such as cephalosporin C and the intermediate 7-aminocephalosporanic acid with ≥98% conversion and 91–93% product yield. |
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Keywords: | 7-Aminocephalosporanic acid Acetyl esterase Antibiotics Cephalosporin-C Orange peel |
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