World Journal of Microbiology and Biotechnology

, Volume 28, Issue 5, pp 2159–2164

Mutations in the translation initiation region of the pac gene resulting in increased levels of activity of penicillin G acylase

  • Özlem Akkaya
  • Saliha İşsever Öztürk
  • Albert Bolhuis
  • Füsun Gümüşel
Original Paper

DOI: 10.1007/s11274-012-1021-6

Cite this article as:
Akkaya, Ö., Öztürk, S.İ., Bolhuis, A. et al. World J Microbiol Biotechnol (2012) 28: 2159. doi:10.1007/s11274-012-1021-6

Abstract

Penicillin G acylase (PA) is an important enzyme used in the industrial production of b-lactam antibiotics. In this study, the effects of mutations in the translation initiation region of the Escherichia coli pac gene, encoding periplasmic PA, were examined. Several mutations led to increased amounts of PA activity, including those that lengthened the spacer region between the ribosome binding site and the ATG start codon, and those with altered codons on positions +2 and +4 relative to the start codon. These results indicated that the wild-type sequence of the pac gene does not provide maximum expression levels and that the strategies applied in this study can be used to improve production of PA in E. coli. Unexpectedly, our study also suggested that translocation of PA was, in contrast to earlier reports, shown not to require the Twin-arginine translocation pathway for transport into the periplasm.

Keywords

Penicillin G acylase pac gene Translation initiation Codon usage Tat pathway 

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Özlem Akkaya
    • 1
  • Saliha İşsever Öztürk
    • 1
  • Albert Bolhuis
    • 2
  • Füsun Gümüşel
    • 1
  1. 1.Department of Molecular Biology and GeneticsGYTE, Gebze Institute of TechnologyKocaeliTurkey
  2. 2.Department of Pharmacy and PharmacologyUniversity of BathBathUK