Adaptive laboratory evolution of ethanologenic @entity15548 tolerant to furfural and XXXX inhibitors.

@entity413 @entity15548 @entity1068 @entity1543
UNASSIGNED: Furfural and @entity1543 from lignocellulosic hydrolysates are the prevalent inhibitors to @entity15548 during cellulosic @entity1068 production. Developing a strain tolerant to furfural or @entity1543 inhibitors is difficul by using rational engineering strategies due to poor understanding of their underlying molecular mechanisms. In this study, strategy of adaptive laboratory evolution (ALE) was used for development of a furfural and @entity1543 -tolerant strain. After three round evolution, four evolved mutants (ZMA7-2, ZMA7-3, ZMF3-2, and ZMF3-3) that showed higher growth capacity were successfully obtained via ALE method. Based on the results of profiling of cell growth, @entity413 utilization, @entity1068 yield, and activity of key enzymes, two desired strains, ZMA7-2 and ZMF3-3, were achieved, which showed higher tolerance under 7  g/l @entity1543 and 3  g/l furfural stress condition. Especially, it is the first report of @entity15548 that could tolerate higher furfural. The best strain, @entity15548 ZMF3-3, has showed 94.84  % theoretical @entity1068 yield under 3-g/l furfural stress condition, and the theoretical @entity1068 yield of ZM4 is only 9.89  %. Our study also demonstrated that ALE method might also be used as a powerful metabolic engineering tool for metabolic engineering in @entity15548 . Furthermore, the two best strains could be used as novel host for further metabolic engineering in cellulosic @entity1068 or future biorefinery. Importantly, the two strains may also be used as novel-tolerant model organisms for the genetic mechanism on the "omics" level, which will provide some useful information for inverse metabolic engineering.
