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Significant efforts are being made to produce biofuels to replace fossil fuels and address the issues caused by global climate change. Due to its potential better conversion efficiency to useable power, decreased emission of pollutants, and high energy density, H 2 is one of the prospective options that are seen as a desirable future clean energy carrier. Although there are numerous technologies available for producing H 2 , this review concentrates on fermentative H 2 production techniques, their drawbacks, and current developments. While being a promising method, fermentative strategies still have several drawbacks, including low H 2 production yields. Many approaches have been used to address these issues; among them, the field of metabolic pathway engineering has made enormous strides. To improve H 2 generation, this paper reviewed and discussed several metabolic pathways and modified strains. As well as the challenges involved in H 2 scale-up from a laboratory setting to a commercial scale. • Biohydrogen production via fermentative methods is reviewed and elaborated. • Metabolic engineering methods are proposed for scaling up fermentative biohydrogen. • Challenges of fermentative biohydrogen production and potential solutions. • Bionanotechnology and modified strains enhanced biohydrogen production. [ABSTRACT FROM AUTHOR]

• Through green synthesis, a new bimetallic licu-ferrite nanospinel was developed. • LiCu-F has a mean particle size of 59.8 nm and a crystallite size of 45.9 nm. • LiCu-F exhibited sufficient antimicrobial activity and can be used in biological applications. • Within 60 min, LiCu-F exhibited bacteriostatic and bactericidal effects on S. aureus and E. coli. • LiCu-F had lower binding energy (−29.16 kcal/mol) for E. coli than for S. aureus (−38.89 kcal/mol). An aqueous carob leaf extract was employed in the current study as a biogenic agent (reducing and capping agents) for the synthesis of novel bimetallic lithium and copper-based nanospinel ferrites (LiCu-F) with antibacterial activity. LiCu-F has a considerable number of surface -OH groups, a BET-specific surface area of 72.5 m2/g, a mean particle size of 59.8 nm, and a crystallinity index of 78.9%, according to the results of the physicochemical characterisation of the ferrite. Within the first 60 min, LiCu-F showed bacteriostatic and bactericidal effects on S. aureus and E. coli based on the time-kill assay. 0.4 mg of LiCu-F demonstrated 67.9% bacteriostatic effects on E. coli while exhibiting 100% inactivation of S. aureus strains, demonstrating that the gram-positive bacteria are more susceptible to LiCu-F. The computational molecular docking analysis demonstrated that LiCu-F has a higher binding affinity for S. aureus (-38.89 kcal/mol) than E. coli (-29.16 kcal/mol), which is consistent with the experimental findings. With an LD 50 of 1000 mg per kg of body weight based on a computational prediction tool, LiCu-F is cytotoxic to bacteria and can cause DNA damage and cell death in bacterial strains. [Display omitted] [ABSTRACT FROM AUTHOR]