With rapidly growing energy demand and concerns over energy security and environment, researchers worldwide are exploring hard to deploy renewable energy sources. Development of economical biofuel at sufficiently large scale may provide major breakthrough in this direction, with strong impact on sustainability. More importantly, environmental benefits may also be achieved by the utilization of renewable biomass resources, which could help the biosphere in longer time. This chapter reviews the availability and bioenergy potentials of the current biomass feedstock. These include the following: (i) food crops such as sugarcane, corn and vegetable oils, classified as the first-generation feedstocks, and environmental and socio-economic barriers limiting its use; (ii) second-generation feedstocks involving lignocellulosic biomass derived from agricultural and forestry residues and municipal waste followed by constraints for their full commercial deployment. Key technical challenges and opportunities of the lignocellulosic biomassto-bioenergy production are discussed in comparison with the first-generation technologies. (iii) The potential of the emerging third-generation biofuel from algal biomass is also reviewed.