From genes to molecules: mining bacterial genomes for new enzyme reactions

有没有可能一把土壤或一桶海水就能解决当今世界面临的一些紧迫的环境和人类健康问题？大规模DNA测序的出现揭示了一个分子世界，它确实可能提供清理污染物的答案或找到下一种神奇药物的途径。受DNA驱动的新化学反应和分子发现的新时代的启发，我们建议探索具有改善环境和人类健康潜力的酶。具体来说，我们将研究碳磷裂解酶的显著的碳磷键断裂反应。我们还将研究PhnY / PhnZ有机膦酸酯降解系统，最近通过筛选海洋DNA，似乎通过一种新的反应机制切割碳-磷键。这两种酶系统都有可能降解环境污染物，如有机膦酸酯，除草剂，杀虫剂，化学战剂和表面活性剂的组成部分。我们还将研究参与“卤化”天然产物生物合成的酶，这是一个具有巨大潜力的生物活性分子大家族，可提供新的抗癌和抗生素药物。具体来说，我们研究了独特的反应催化的“卤化酶”参与的生物合成的第一个神奇的药物之一的世纪，抗生素氯霉素。利用从基因组测序获得的信息，我们还将描述杀核素的生物合成，杀核素是一种罕见的氟取代天然产物。结合氟原子的酶的鉴定，一个“氟化酶”，可能导致用于合成含氟核苷的生物催化剂，这是作为抗病毒抑制剂的极大兴趣。总的来说，我们的建议包含4个研究领域，将培养4名研究生，以及每年的暑期学生，从分子生物学和微生物学到合成化学的各种技能。这些技能在制药和生物技术公司日益多学科的环境中需求量很大。

Is it possible that a handful of soil or a pail of seawater could hold the solution to some of the pressing environmental and human health issues facing the world today? The advent of large scale DNA sequencing has revealed a molecular world that may indeed provide the answer to cleaning up pollutants or the path to the next wonder drug. Inspired by this new era of DNA driven discovery of new chemical reactions and molecules, we propose to explore enzymes that have the potential to improve environmental and human health. Specifically, we will study the remarkable carbon-phosphorus bond cleaving reaction of carbon-phosphorus lyase. We will also study the PhnY / PhnZ organophosphonate degradation system, recently identified through screening of marine DNA, which appears to cleave carbon-phosphorus bonds through a new reaction mechanism. Both enzyme systems have the potential to degrade environmental pollutants known as organophosphonates, a component of herbicides, insecticides, chemical warfare agents, and surfactants. We will also study enzymes that are involved in the biosynthesis of 'halogenated' natural products, a large family of bioactive molecules that have great potential to provide new anticancer and antibiotic drugs. Specifically we examine the unique reaction catalyzed by the 'halogenase' involved in the biosynthesis of one of the first wonder drugs of the 20th century, the antibiotic chloramphenicol. Using information gained from genome sequencing, we will also characterize the biosynthesis of, nucleocidin, a rare example of a fluorine substituted natural product. Identification of the enzyme that incorporates the fluorine atom, a 'fluorinase', could lead to a biocatalyst for the synthesis of fluorine containing nucleosides, which are of great interest as antiviral inhibitors. Overall our proposal contains 4 areas of study that will train 4 graduate students, as well as a yearly summer student, in a variety of skills, from molecular biology and microbiology to synthetic chemistry. These skills are in high demand in the increasingly multidisciplinary environment that exists in pharmaceutical and biotechnology companies.