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Rivista di Biologia Molecolare e Biotecnologie

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Minerva Biotecnologica 2014 December;26(4):263-79

lingua: Inglese

Fermentative optimization of culture conditions for large scale production of engineered synthetic antimicrobial peptide using salt inducible E. coli GJ1158

Seetha Ram K. 1, Pulicherla K. K. 2, Peravali J. B. 3

1 Department of Biotechnology, Acharya Nagarjuna University, Nagarjuna Nagar Guntur, Andhra Pradesh, India;
2 CBST, VIT, Vellore, TN, India;
3 Department of Biotechnology, Bapatla Engineering College, Bapatla Guntur, Andhra Pradesh, India


AIM: The aim of this paper was to optimize the media composition, influence of components and culture conditions using one-factor-at-a-atime, statistical designs and batch and fed batch fermentation for the production of synthetic cationic antimicrobial peptide (synthetic cAMP) from salt inducible E. coli GJ1158.
METHODS: Best medium constituents are optimized by one-factor-at-a-time. Statistical designs like Taguchi and response surface methodology are used to enhance the production of synthetic cationic antimicrobial peptide from E. coli GJ1158. Further large scale production was carried out using batch and fed batch fermentation. In fermentation the effect of individual components are also studied.
RESULTS: In one-factor-at-a-time, the best medium constituents viz., carbon (glucose), nitrogen (yeast extract), phosphate sources (disodium hydrogen phosphate) and culture conditions i.e., pH (7), temperature (37 °C) and inducer concentration (100 mM NaCl) were identified. The L27 orthogonal array expressed 142 mg/L of recombinant synthetic cAMP. Final optimization is carried out with the response surface methodology yields 168 mg/L of synthetic cAMP. Later batch fermentation with LBON and MGYEON yields 248 mg/L and 201 mg/L. Fed batch fermentation trails results the maximum quantity i.e., 467 mg/L of recombinant synthetic cationic antimicrobial peptide on glucose, yeast extract and Na2HPO4 as pulse at 40% dissolved oxygen concentraion (DOC).
CONCLUSION: Synthetic cationic AMP are the better choice to existing antibiotics. For the first time, in this study production of synthetic cationic AMP was enhanced using the one-factor-at-a-time and statistical methods (Taguchi and RSM) and further fermentation studies are conducted for the large scale production of antimicrobial peptide from E. coli GJ1158.

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