High-yield production of active recombinant S. simulans lysostaphin expressed in E. coli in a laboratory bioreactor

• Published in: Protein expression and purification 2021-01, Vol.177, p.105753-105753, Article 105753
• Main Authors: Duman-Özdamar, Zeynep Efsun, Ünlü, Aişe, Ünal, Hayriye, Woodley, John M., Bi̇nay, Barış
• Format: Article
• Language: English
• Subjects: Auto-induction medium; E. coli expression system; L- (+) arabinose; Laboratory bioreactor; Lysostaphin; Scale-up
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2020.105753

Staphylococcus aureus (S. aureus), which has developed multidrug resistance, leads to many healthcare-associated infections resulting in significant medical and economic losses. Therefore, the development of new efficient strategies to deal with these bacteria has been gaining importance. Lysostaphin is a peptidoglycan hydrolase that has considerable potential as a bacteriocin. However, there have been few reported optimization and scale-up studies of the lysostaphin bioproduction process. Our preliminary results have revealed that the composition of auto-induction media at 30 °C increases the produced lysostaphin around 10-fold in shake flasks. In this study, achieving higher yields for recombinant lysostaphin in E. coli at a laboratory scale has been the aim, through the use of auto-induction media. Optimized medium composition and fermentation parameters were transferred to a laboratory-scale bioreactor. The tested conditions improved protein yields up to 184 mg/L in a 3 L stirred bioreactor and the productivity was improved 2-fold in comparison to previously published reports. Furthermore, this study also showed that lysostaphin is an effective bacteriocin on both commercially available and isolated S. aureus strains. These results will contribute to future larger-scale production of lysostaphin via the proposed fermentation conditions. •Produced increased amounts of lysostaphin in an active form using auto induction in E. coli in a laboratory bioreactor.•Investigation of the effect of agitation speed and type of carbon source on production yield.•Development of a suitable procedure for larger-scale production of lysostaphin.