Annals of Microbiology

, Volume 62, Issue 3, pp 1287–1293 | Cite as

Characterization of a new Bacillus cereus ATUAVP1846 strain producing penicillin V acylase, and optimization of fermentation parameters

  • Avinash Vellore Sunder
  • Atul Kumar
  • Neha Naik
  • Archana V. Pundle
Original Article
First Online:
Received:
Accepted:

Abstract

Penicillin acylases are involved chiefly in the industrial production of semisynthetic penicillins, which remain the most widely used group of antibiotics. We have isolated a new bacterial strain ATUAVP1846 that produces penicillin V acylase (PVA). Phylogenetic analysis using 16S rRNA sequencing showed 99.37% homology with Bacillus cereus. Maximum PVA production was observed with B. cereus ATUAVP1846 at 30°C, pH 7 after 24 h fermentation time under submerged conditions. Highest enzyme productivity was achieved using sucrose as carbon source, and tryptone and ammonium hydrogen phosphate as nitrogen sources. Minimal medium containing 0.4% glucose and 0.3% ammonium hydrogen phosphate was found to be optimal for maximum PVA production from B. cereus ATUAVP1846. The crude enzyme from B. cereus ATUAVP1846 was partially purified using ammonium sulfate fractionation and showed highest enzymatic activity in the hydrolysis of penicillin V at 40°C and pH 6. The crude enzyme preparation also showed unique substrate specificity, preferring ampicillin and cephalexin over penicillin V.

Keywords

Penicillin acylase Bacillus cereus 16S rRNA Isolation Ntn hydrolase Optimization 
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Notes

Acknowledgments

Avinash V.S. thanks the Council of Scientific and Industrial Research (CSIR), India for the award of Junior Research Fellowship (JRF). The authors thank Dr. Tanuja Gupte, Hindustan Antibiotics, Pune, India for providing Penicillin V potassium salt.

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Copyright information

© Springer-Verlag and the University of Milan 2011

Authors and Affiliations

  • Avinash Vellore Sunder
    • 1
  • Atul Kumar
    • 2
  • Neha Naik
    • 3
  • Archana V. Pundle
    • 1
  1. 1.Division of Biochemical SciencesNational Chemical LaboratoryPuneIndia
  2. 2.Unite of Rickettsies (WHO Collaborative Centre)University of the MediterraneanAix-MarseilleFrance
  3. 3.Modern CollegePuneIndia

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