S58: Promises and challenges of applying next-generation sequencing techniques for robust biocatalyst development

A key barrier for economic biofuel production is the development and deployment of robust biocatalysts capable of utilizing non-native substrates with high productivities and yields. Rapid technology progress in next-generation sequencing (NGS) changes the paradigm and strategies for biocatalyst development making the understanding of the biocatalysts at a global level feasible. Using Zymomonas mobilis as a model system, we will discuss our efforts to better understand biomass pretreatment hydrolysate inhibitor tolerance and nonnative sugar utilization using NGS approaches, and insights we have obtained from these studies. Specifically, NGS-based genome resequencing has been applied to investigate the genetic differences among strains developed through mutagenesis and lab-directed adaptive evolution. We also use transcriptomics of chip-based high-density microarray and NGS-based strand specific RNA-Seq (ssRNA-Seq) to study the stress responses of Z. mobilis to the major inhibitors when grown on different carbon sources. Additionally, we will discuss the technical challenges we experienced when applying these NGS-based techniques for biocatalyst development and potential solutions to address them.