OPTIMIZATION OF A CULTURE MEDIUM FOR BIOMASS AND δ-ENDOTOXIN PRODUCTION BY A RECOMBINANT ESCHERICHIA COLI STRAIN

Document Type : Original Article

Authors

1 Dept. Microbiol., Fac. Agric., Ain Shams University, Shobra El-Khema, Cairo, Egypt

2 Plant Protection Research Institute, Agric. Research Center, Dokki, Giza, Egypt

Abstract

A recombinant strain of Escherichia coli harboring a plasmid containing the Bacillus thuringiensis δ-endotoxin synthesis gene, was tested for its efficacy to synthesize δ-endotoxin, in a complex medium containing sucrose and yeast extract. Also, the recombinant E. coli strain was tested for its efficacy against the 2nd instars of Spodoptera littoralis. The recombinant strain of E. coli showed a good activity against the 2nd instars of S. littoralis, the mortality was 70 % after 7 days at room temperature. A high cell biomass (8.8gL-1) and δ-endotoxin concentration (6.8 mg L-1), were obtained by the shake flask culture (100 ml medium/250 ml flask, at 200 rpm), of the recombinant E. coli in modified MR medium containing sucrose (20g/L), as carbon source and yeast extract as nitrogen source, in the presence of CaCO3, K2HPO4, MgSO4, FeSO4 and ZnSO4 as mineral salts. The best pH values for cell biomass production and endotoxin production were 7.0 and 7.5, respectively. The corresponding figures for the best temperature were 37oC and 30oC, respectively. The use of some byproducts such as black-strap molasses, corn-steep liquor and cheese whey, as an alternative for carbon and nitrogen sources of medium, were found to enhance the cell growth but showed no effect on endotoxin production.

Keywords

Files

Share

How to cite

Statistics