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Recent years have seen an explosion in the use of advanced biotechnology techniques in academic and industrial activities to modulate microorganism pathways for the production of fuels or chemicals. Synthetic biology is adopted for biofuel production, and it needs scientific evidence to support the fundamentals and risk assessments. Biofuel is derived from biomass of a plant, animal waste, or microalgae. These materials can be replenished after some time; hence they are a renewable source of energy. Risk assessment is considered significant to maintain and comply with regulatory frameworks existing around the world. The use of scientific tools such as enzymes and microbes itself needs review and approval. Risk profiles are done for the toxicity, infectivity, and strategies. Recent years have seen an exciting increase in developing strategies for the use of advanced biotechnology techniques to enhance the …