Highly active and stable biocatalysts are the prerequisite for industrial scale application of the biodesulfurization process. Scientists are making efforts for increasing the desulfurizing activity of native strains by employing various genetic engineering approaches. Nevertheless, the achieved desulfurization rate is lower than the industrial requirements. Thus, there is a dire need to use efficient genetic tools for precise genome editing of desulfurizing bacteria for enhanced efficiency. In comparison to the previously used genetic engineering tools the newly developed CRISPR-Cas is a more efficient and simple genetic tool that has been successfully applied for targeted genome modification of eukaryotes as well as prokaryotes. In this paper, we have reviewed the approaches, previously used to enhance the biodesulfurization rates of the sulfur metabolizing microorganisms and have discussed the potential of CRISPR-Cas systems in engineering desulfurizing biocatalysts. We have also proposed a model to construct competent desulfurizing recombinants involving use of CRISPR-Cas technology. The model can be used to over-express the dsz genes under a constitutive promoter in a suitable heterologous host, to get a steady expression of desulfurization pathway. This may serve as an inducement to develop better performing desulfurizing recombinant strains using CRISPR-Cas systems, which can be helpful in increasing the rate of biodesulfurization in future.

高活性和稳定的生物催化剂是生物脱硫工艺工业规模应用的前提。科学家们正在努力通过采用各种基因工程方法来提高天然菌株的脱硫活性。然而，获得的脱硫率低于工业要求。因此，迫切需要使用有效的遗传工具对脱硫细菌进行精确的基因组编辑以提高效率。与以前使用的基因工程工具相比，新开发的CRISPR-Cas是一种更有效，更简单的遗传工具，已成功应用于真核生物和原核生物的靶向基因组修饰。在本文中，我们回顾了这些方法，以前用于提高硫代谢微生物的生物脱硫速率，并讨论了CRISPR-Cas系统在工程脱硫生物催化剂中的潜力。我们还提出了一种使用CRISPR-Cas技术构建胜任脱硫重组体的模型。该模型可用于过分表达dsz基因在适当的异源宿主中的组成型启动子下获得稳定的脱硫途径表达。这可能是使用CRISPR-Cas系统诱使开发性能更好的脱硫重组菌株的诱因，这可能有助于将来提高生物脱硫率。