This book is presented on biofuel production which includes different technologies developed and adopted to synthesize green renewable fuel alternatives for sustainable development. It also reflects different sources of biofuel, application of microbial community and microbial engineering to design fuel production and the biosynthetic pathways of biofuel production by microbes. Although the expenses for the physical and chemical technologies for energy production and fossil fuel utilization to protect our environment are very high, these technologies are not eco-friendly and safe. Hence, the need of certain modern eco-friendly and cost-effective techniques to protect our environment is deeply apprehended by different workers of this field. These techniques involve some feasible technologies utilizing different biological agents like microbes to produce renewable energy. This book provides an outline of the science behind the multidisciplinary aspects of biofuel production. It summarizes a solid foundation in the fundamentals and progresses to practical applications in this field. It structures stepwise route for a number of effective techniques to screen, select, identify and utilize microbes for biofuel production and utilization. It also focuses on the theoretical groundworks of biofuel production, recent technologies related to microbial engineering like myco-engineering technologies, microbial metabolism or modelling approaches to microbial physiology utilized for the same purpose. The techniques covered in this book ensure that scientists have the knowledge to practice effective biofuel production techniques themselves in a contaminated ecosystem in a sustainable way. Recent progress in the field of biofuels using microbial genetic engineering has larger perspectives in commercial-scale production. However, its large-scale production is still challenging; hence, to resolve this problem, it is essential to convert biomass into biofuels by developing novel technology to increase biofuel production to fulfil the current and future energy demand.