Molecular Insight into Toxicant Degradation by Microbial Communities

描述（由申请人提供）：微生物具有很大的生物化学潜力，能够利用天然和人为化合物作为生长和能源。它们对极端环境条件具有独特的适应性，在地球上随处可见。然而，众所周知，微生物的生物多样性仍然相对未开发，环境中超过99%的细菌未被鉴定。烷烃降解细菌是用于研究微生物生物多样性的示例模型系统，从环境修复和工业生物催化的观点来看，具有工业相关性的额外益处：烷烃降解细菌在石油的生物降解中具有活性，并且它们的烷烃氧化酶在进行药物相关性的手性转化中是有价值的。从海洋、极地、土壤和污染地点分离出独特的烷烃降解细菌。许多新的属和种最近已被描述的基础上分离的有机体的能力，降解烷烃。吉尔吉斯共和国是世界上生境多样的一个地区，有高山沙漠、常年积雪的山脉、盐碱土壤、前苏联时期遗留下来的石油和重金属污染区、农田、山林和溪流、原始山谷和一个大的（100英里长，40英里宽）微咸的山湖（伊塞克湖）。这些环境尚未系统地检查它们所拥有的微生物的多样性。本提案旨在探索吉尔吉斯共和国烷烃降解细菌的微生物多样性，目的是发现和鉴定吉尔吉斯共和国各种生境所特有的细菌和烷烃羟化酶基因。将采取分子生态学方法，使用各种方法来检查直接从环境和富集培养物中提取的DNA。微生物多样性将使用16S rRNA基因作为生物标志物进行监测，烷烃降解多样性将使用二铁和细胞色素P450烷烃单加氧酶基因作为靶标进行检查。该提案汇集了吉尔吉斯共和国土壤微生物学科学家与罗格斯大学的教师，他们在分解代谢途径的分子表征方面拥有丰富的经验。燃料碳氢化合物如烷烃是环境中常见的污染物。接触这些化合物会影响人类健康和福祉。在吉尔吉斯斯坦共和国等发展中国家和偏远国家，了解与这些化学品的补救有关的微生物及其分子生态学，在确定这些和其他有毒化合物的环境归宿方面发挥着重要作用。将通过这项拟议研究的互动活动，提高吉尔吉斯共和国人民对污染危害的认识。

DESCRIPTION (provided by applicant): Microorganisms have a large biochemical potential, with the capability to use natural and anthropogenic compounds as growth and energy sources. They possess unique adaptability to extreme environmental conditions and are found everywhere on the Earth. However, it is well known that the biodiversity of microorganisms remains relatively untapped with more than 99% of all bacteria in the environment uncharacterized. The alkane degrading bacteria are example model systems for studying microbial biodiversity with the added benefit of industrial relevance from both the environmental remediation and industrial biocatalysis standpoints: alkane degrading bacteria are active in the biodegradation of petroleum and their alkane oxidizing enzymes are valuable in performing chiral transformations of pharmaceutical relevance. Unique alkane degrading bacteria have been isolated from the ocean, from the Polar Regions, from soil, and from polluted sites. Many new genera and species have recently been described based on organisms isolated for the ability to degrade alkanes. The Kyrgyz Republic represents a region of the world with varied habitats ranging from high mountain deserts, permanently snow-capped mountains, heavily saline and alkaline soils, petroleum and heavy metal polluted sites left over from the soviet era, agricultural fields, mountain forests and streams, pristine valleys, and a large (100 miles long by 40 wide) mildly saline mountain lake (Lake Issyk-Kyl). These environments have not yet been systematically examined for the diversity of microorganisms that they possess. The present proposal aims to explore the microbial diversity of alkane degrading bacteria in the Kyrgyz Republic with the aim of finding and characterizing bacteria and alkane hydroxylase genes that are unique to the varied habitats in the Kyrgyz Republic. A molecular ecology approach will be taken, using a variety of methodologies to examine DNA directly extracted from the environment and from enrichment cultures. Microbial diversity will be monitored using the 16S rRNA gene as a biomarker and alkane degrading diversity will be examined using diiron and cytochrome P450 alkane monooxygenase genes as the target. The proposal brings together Kyrgyz Republic soil microbiology scientists with faculty at Rutgers University who have extensive experience in the molecular characterization of catabolic pathways. Fuel hydrocarbons such as alkanes are common contaminants in the environment. Exposure to these compounds can affect human health and well being. Knowledge of the microorganisms which are involved in the remediation of these chemicals and their molecular ecology in developing and remote countries like the Kyrgyz Republic plays a major role in determining the environmental fate of these and other toxic compounds. Public awareness among the people of the Kyrgyz Republic of the hazards of pollution will be raised through the interactive activities of this proposed research.