Optimization of Escherichia coli Bacteriophages Production
V. D. Hari Narayana Kola
Department of Microbiology, Nizam College, Osmania University, Hyderabad, Telangana, India and Department of Biotechnology, Vignan’s Foundation for Science Technology and Research, Deemed to be University, Vadlamudi, Guntur District, Andhra Pradesh, India.
Koigoora Srikanth
Department of Biotechnology, Vignan’s Foundation for Science Technology and Research, Deemed to be University, Vadlamudi, Guntur District, Andhra Pradesh, India.
Shaik Muzammil Pasha
Department of Microbiology, Nizam College, Osmania University, Hyderabad, Telangana, India.
Y. Sudhan Goutham
Department of Microbiology, Nizam College, Osmania University, Hyderabad, Telangana, India.
Chand Pasha *
Department of Microbiology, Nizam College, Osmania University, Hyderabad, Telangana, India.
*Author to whom correspondence should be addressed.
Abstract
Aims: Phage therapy may be the best alternative to antibiotics in poultry. There is a need for optimization of phage production conditions to produce more Escherichia coli specific phages for poultry applications.
Methodology: An E. coli λ bacteriophage isolated in our laboratory was used for optimization studies. An optimization of the E. coli λ bacteriophage production process was carried out by conventional methods to identify the factors, and the Thaguchi process was used for the optimization of conditions for increased bacteriophage production. With four factors each at three levels L9, experimental runs were carried out.
Results: In the conventional method, 8 factors were studied and 4 selected for statistical optimization by Taguchi. At optimized conditions of modified nutrient broth with 1% glycerol, temperature 37oC, 100 rpm for 12 hours of phage multiplication, an improvement of the final titer was achieved from a control 2x109 to optimized 5x1012 Plauque formation Units (PFU)/mL.
Conclusion: Phage production is dependent on bacterial growth and bacteriophage multiplication in bacteria. In such a complicated system, the Thaguchi method will be a convenient method of phage production optimization.
Keywords: Bacteriophage, E. coli, phage therapy, λ Phage, Taguchi