Collaborative Research: Biotransformations Near and Above 100?C: Mining Genomes of Hyperthermophiles for New Biocatalysts

生物催化剂的发现和表征仍然是一个关键的生物技术目标。在过去的几年里，高通量筛选和定向进化方法已经发展起来，使这些生物催化剂能够被识别，然后根据它们的生化、生物物理和生物处理特性进行优化。然而，目前的发现和优化方法严重依赖于基于酶的测定，由于化学限制或基因表达问题，这可能很难甚至不可行。此外，由于目前的方法侧重于单个基因的产物，因此这种方法不容易识别多酶转化。作为规避现有方法局限性的一种手段，本研究的目标是开发基于功能基因组学的发现方法来鉴定新的生物催化剂。随着测量基因表达的高通量技术的出现，细胞在遗传水平上的反应可以与环境条件相关联。这种酶发现的替代方法利用了高度敏感的转录。基于细胞对生物技术相关的环境挑战和生长条件的反应。在本项目期间，将广泛使用DNA微阵列来识别编码新酶的基因，以进行一系列重要的生物转化。

Biocatalyst discovery and characterization remains a key biotechnological objective. High throughput screening and directed evolution approaches have been developed over the last few years that enable such biocatalysts to be identified and then optimized with respect to their biochemical, biophysical and bioprocessing properties. However, current methods for discovery and optimization rely heavily on enzyme-based assays, which may be difficult or even infeasible to implement because of chemical limitations or gene expression issues. Moreover, multienzyme transformations are not easily identified by this approach since current methods focus on the products of individual genes. As a means of circumventing the limitations with current approaches, the objective of this research is to develop functional genomics-based discovery methods to identify novel biocatalysts. With the advent of high-throughput technologies to measure gene expression, cellular responses at the genetic level can be correlated to environmental conditions. This alternative approach to enzyme discovery takes advantage of highly sensitive, transcriptional.based cellular responses to biotechnologically-related environmental challenges and growth conditions. In this project period, extensive use will be made of DNA microarrays to identify the genes that encode new enzymes for a range of important biotransformations.