DESIGNED ORF PROTEIN FOR SUSTAINABLE THIRD GENERATION BIOFUEL PRODUCTION

The increasing global energy demands and depletion of fossil fuels urge us to develop alternative energy resources. In recent years, algae has gained much attention as a third-generation biofuel feedstock. Among Algae, microalgae are considered a good source of biofuels because of their relatively more oil concentration and rapid production of biomass. Microalgae have the ability to produce a wide variety of biofuels including bio-oil, bio-diesel, bio-syngas, and bio-hydrogen. Commercial production of biofuels from microalgae is presently not in use because of the high expense of production. This research was designed to produce a hybrid ORF that might enhance the oil yielding ability of the microalgae by its translation into a working protein. Ban y using an intensive literature survey, 6 genes from 3 microalgae species were selected. Further, ORFs of selected genes were identified and Hybrid ORF was constructed by combining those ORFs. After that restriction enzyme analysis and thermodynamic analysis of the hybrid nucleotide sequence was carried out, and this showed that the hybrid sequence was stable. The Hybrid ORF nucleotide sequence was translated into the protein sequence. This protein sequence was used for the homology modeling of Hybrid ORF proteins. The protein conformation predicted by homology modeling was further verified by Ramachandran plot. The metabolic pathways were analyzed by KEGG, which showed that all selected genes were functioning in the biosynthesis of lipids. This showed that the Hybrid ORF constructed could be used in a way to enhance the oil content of microalgae for producing biofuel, particularly biodiesel. These results can be validated by further in vitro analysis and can be used for the construction of genetically engineered microalgae.