Novel enzymes for conversion of biomass to energy: applied structural and functional proteomics of uncharacterized microbial hydrolases and oxidoreductases

当亨利·福特在1896年制造他的第一辆汽车时，他把它设计成使用乙醇。在德国，鲁道夫柴油公司制造的这款发动机使用花生油。然而，随着汽车的兴起，这些可再生的生物燃料很快被石油取代。目前，高能源价格、能源进口增加、对石油供应的担忧以及对化石燃料环境后果的更多认识推动了人们对生物燃料和生物质转化为能源的兴趣。如今，生物柴油、生物乙醇和生物氢等几种基于酶的可再生和清洁能源生产技术正在迅速崛起。今天，生物能源约占世界能源消费的15%，其产量正在快速增长。尽管生物催化有明显的好处，但仍有几个主要障碍阻碍了这些技术的实施：生产成本高，产率低，酶活性和稳定性低，可用酶库非常有限。全球基因组和元基因组测序工作发现了数百万编码新的未知蛋白质的基因。它们代表着将生物质转化为能源的新酶的潜在丰富来源。该项目的主要目的是发现对生物柴油生产和植物生物质降解具有潜在重要意义的新型微生物酶。我们建议应用我们的高通量酶筛选和结构蛋白质组学技术来鉴定和快速鉴定100种新的酶，重点放在脂肪酶和酯酶上，了解这些酶的底物选择性和稳定性的分子基础，最终选择几种未来应用于生物能源的酶。

When Henry Ford built his first car in 1896, he designed it to run on ethanol. In Germany, Rudolph Diesel builthis engines to run on peanut oil. However, with the rise of the automobile these renewable biofuels werequickly replaced by petroleum. Presently, high energy prices, increased energy imports, concerns aboutpetroleum supplies, and greater recognition of the environmental consequences of fossil fuels have driveninterest in biofuels and conversion of biomass to energy. Nowadays, several enzyme-based technologies for theproduction of renewable and clean energy sources, such as biodiesel, bioethanol and biohydrogen, are rapidlyemerging. Today, bioenergy accounts for approximately 15% of the energy consumption of the world and itsproduction is quickly growing. Despite the obvious benefits of biocatalysis, there are several major hurdleshindering the implementation of these technologies: high production costs, low yields, low enzyme activity andstability, and very limited repertoire of available enzymes.Global genome and metagenome sequencing efforts uncovered millions of genes encoding novel, unknownproteins. They represent a potentially rich source of new enzymes for the conversion of biomass to energy. Themain aim of this project is to discover novel microbial enzymes of potential importance for biodieselproduction and degradation of plant biomass. We propose to apply our high-throughput technologies ofenzymatic screening and structural proteomics to identify and quickly characterize 100 novel enzymes with afocus on lipases and esterases, to understand the molecular basis of substrate selectivity and stability of theseenzymes, and eventually, to select several enzymes for future application in bioenergy.