Assessment of Selected Industrial Byproducts as Alternative Nutrient Source for E. Сoli pET30a-HIV-RT

The cost and availability of culture media can significantly impact the economic feasibility of recombinant protein technology. Thus, this study examines the potential of industrial byproducts as an alternative nutrient source for E. coli pET30a-HIV-RT, which was designed to produce the viral enzyme Reverse Transcriptase. The growth effects of the industrial byproducts were assessed based on optical densities (OD 600 ) and generation time. Here, we show that among the extracts from four selected industrial byproducts, comparable OD 600 was observed in sugarcane molasses, Brewer’s yeast, and soybean flour. Since residues in Brewer’s yeast interferes with OD 600 measurement, we selected sugarcane molasses (SM-M9) at 1.35 g/L and soybean flour (SBF-M9) at 3.67 g/L as alternative carbon and nitrogen sources, respectively, in the M9 minimal media. Compared to SBF-M9 (63.19±7.67 min), a shorter generation time was observed in SM-M9 (44.60±3.67 min), and this is comparable with the generation time observed in Luria-Bertani broth (36.85±0.05) and M9 (47.95±3.02 min). On the other hand, the OD 600 observed in SBF-M9 (1.17±0.05) is comparable with M9 (1.23±0.02). Overall, our results indicate the potential of molasses and soybean flour to serve as alternative nutrient sources for genetically engineered E. coli. Future research will focus on the assessment of the effects of molasses and soybean flour on recombinant Reverse Transcriptase expression and purification.