Geopolitical and economic factors have motivated the scientific community to utilize renewable energy resources. In addition to the modifications in major steps and processes of biofuel production, manipulation of microbial genome engineering tools is essential in order to find sustainable solution of continuous depletion of fossil-fuels. Recently, the CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (CRISPR-associated nuclease 9), a prokaryotic molecular immunity system, has emerged as a novel technology for targeted genomic engineering. This genetic machinery seems to be a groundbreaking discovery to engineer the microbial genomes for desired traits such as enhancing the biofuel tolerance, inhibitor tolerance and thermotolerance as well as modifying the cellulases and hemicelluloses enzymes. In this review, a summary of different generations of biofuels, integrated processes of bioconversion of raw materials into biofuels and role of microbes in biofuel production has been presented. However, the ultimate focus of the review is on major discoveries of CRISPR/Cas9-mediated genome editing in microorganisms and exploitation of these discoveries for enhanced biofuel production.

地缘政治和经济因素促使科学界利用可再生能源。除了对生物燃料生产的主要步骤和过程进行修改之外，微生物基因组工程工具的操纵对于找到持续消耗化石燃料的可持续解决方案也是至关重要的。最近，一种原核分子免疫系统CRISPR（聚簇的规则间隔的短回文重复序列）/ Cas9（CRISPR相关的核酸酶9）已经成为一种靶向基因组工程的新技术。这种遗传机制似乎是一项突破性的发现，旨在为所需的性状改造微生物基因组，例如增强生物燃料耐受性，抑制剂耐受性和耐热性以及修饰纤维素酶和半纤维素酶。在这篇评论中，概述了不同世代的生物燃料，从原材料到生物燃料的生物转化的综合过程以及微生物在生物燃料生产中的作用。然而，该综述的最终重点是在微生物中CRISPR / Cas9介导的基因组编辑中的重大发现，以及对这些发现的利用，以增强生物燃料的生产。