Amores Gerardo Ruiz, Guazzaroni María-Eugenia, Arruda Letícia Magalhães, Silva-Rocha Rafael
FMRP - University of São Paulo, Ribeirao Preto, SP, Brazil;
FFCLRP - University of São Paulo, Ribeirao Preto, SP, Brazil.
Curr Genomics. 2016 Apr;17(2):85-98. doi: 10.2174/1389202917666151116212255.
Filamentous fungi are remarkable organisms naturally specialized in deconstructing plant biomass and this feature has a tremendous potential for biofuel production from renewable sources. The past decades have been marked by a remarkable progress in the genetic engineering of fungi to generate industry-compatible strains needed for some biotech applications. In this sense, progress in this field has been marked by the utilization of high-throughput techniques to gain deep understanding of the molecular machinery controlling the physiology of these organisms, starting thus the Systems Biology era of fungi. Additionally, genetic engineering has been extensively applied to modify wellcharacterized promoters in order to construct new expression systems with enhanced performance under the conditions of interest. In this review, we discuss some aspects related to significant progress in the understating and engineering of fungi for biotechnological applications, with special focus on the construction of synthetic promoters and circuits in organisms relevant for industry. Different engineering approaches are shown, and their potential and limitations for the construction of complex synthetic circuits in these organisms are examined. Finally, we discuss the impact of engineered promoter architecture in the single-cell behavior of the system, an often-neglected relationship with a tremendous impact in the final performance of the process of interest. We expect to provide here some new directions to drive future research directed to the construction of high-performance, engineered fungal strains working as microbial cell factories.
丝状真菌是天然擅长解构植物生物质的非凡生物体，这一特性在利用可再生资源生产生物燃料方面具有巨大潜力。在过去几十年里，真菌基因工程取得了显著进展，以培育出一些生物技术应用所需的与工业兼容的菌株。从这个意义上说，该领域的进展以利用高通量技术深入了解控制这些生物体生理的分子机制为标志，从而开启了真菌系统生物学时代。此外，基因工程已被广泛应用于修饰特征明确的启动子，以便构建在感兴趣条件下具有增强性能的新表达系统。在本综述中，我们讨论了与真菌在生物技术应用中的理解和工程方面的重大进展相关的一些方面，特别关注与工业相关生物体中合成启动子和电路的构建。展示了不同的工程方法，并研究了它们在这些生物体中构建复杂合成电路的潜力和局限性。最后，我们讨论了工程化启动子结构对系统单细胞行为的影响，这是一种常常被忽视但对感兴趣过程的最终性能有巨大影响的关系。我们期望在此提供一些新方向，以推动未来针对构建作为微生物细胞工厂的高性能工程真菌菌株的研究。
