title: Biotransformation of penicillin V to 6-aminopenicillanic acid using immobilized whole cells of E. coli expressing a highly active penicillin V acylase.
abstract: The production of 6-aminopenicillanic acid (6-APA) is a key step in the manufacture of semisynthetic antibiotics in the pharmaceutical industry. The penicillin G acylase from Escherichia coli has long been utilized for this purpose. However, the use of penicillin V acylases (PVA) presents some advantages including better stability and higher conversion rates. The industrial application of PVAs has so far been limited due to the nonavailability of suitable bacterial strains and cost issues. In this study, whole-cell immobilization of a recombinant PVA enzyme from Pectobacterium atrosepticum expressed in E. coli was performed. Membrane permeabilization with detergent was used to enhance the cell-bound PVA activity, and the cells were encapsulated in calcium alginate beads and cross-linked with glutaraldehyde. Optimization of parameters for the biotransformation by immobilized cells showed that full conversion of pen V to 6-APA could be achieved within 1 hr at pH 5.0 and 35°C, till 4% (w/v) concentration of the substrate. The beads could be stored for 28 days at 4°C with minimal loss in activity and were reusable up to 10 cycles with 1-hr hardening in CaCl2between each cycle. The high enzyme productivity of the PVA enzyme system makes a promising case for its application for 6-APA production in the industry.
authors.affiliation: a Division of Biochemical Sciences , CSIR-National Chemical Laboratory , Pune , India.
authors.forename: Vellore Sunder
authors.initials: VS
authors.lastname: Avinash
authors.affiliation: a Division of Biochemical Sciences , CSIR-National Chemical Laboratory , Pune , India.
authors.forename: Palna Dinesh
authors.initials: PD
authors.lastname: Chauhan
authors.affiliation: a Division of Biochemical Sciences , CSIR-National Chemical Laboratory , Pune , India.
authors.forename: Shraddha
authors.initials: S
authors.lastname: Gaikwad
authors.affiliation: a Division of Biochemical Sciences , CSIR-National Chemical Laboratory , Pune , India.
authors.forename: Archana
authors.initials: A
authors.lastname: Pundle
mesh_terms: D000464:Alginates / Q000737:chemistry; D001711:Biotransformation; D004926:Escherichia coli / Q000235:genetics*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D010397:Penicillanic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D010399:Penicillin Amidase / Q000235:genetics / Q000378:metabolism*; D010404:Penicillin V / Q000493:pharmacokinetics*; D010539:Permeability
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D010397:Penicillanic Acid; D020723:Glucuronic Acid; D010399:Penicillin Amidase; C004408:aminopenicillanic acid; D010404:Penicillin V
keywords: Acylase; alginate; biotransformation; immobilization; penicillin; whole cell