Isolating novel fungi for antibiotic discovery

描述(申请人提供)：该项目的长期目标是开发对临床重要的多重耐药(MDR)细菌病原体(如甲氧西林耐药金黄色葡萄球菌(MRSA))和高度耐药的革兰氏阴性病原体(如金黄色假单胞菌)有效的新型抗生素。获得新型抗生素的目标将通过使用一种新开发的方法(真菌捕捉器)来实现，将以前未培养的微真菌作为新抗生素分子的来源进行分离和培养。今天使用的大多数抗生素都是从环境微生物中发现的，包括微真菌。然而，已知的微生物衍生抗生素只从可用标准技术培养的各种微生物中的一小部分(1%)中被发现。微真菌是许多用于治疗疾病的有价值药物的来源，如细菌感染(如头孢菌素)、真菌感染(如蜡烛素)、高胆固醇血症(他汀类药物)和炎症性疾病(如环孢菌素)。全球抗生素市场规模超过200亿美元。对多药耐药病原体非常需要的抗生素有可能产生巨大的市场机会。在这个一期工程中，我们将评估和实施真菌捕捉法分离难以培养的环境微型真菌，并产生一个原理证明，使用该方法培养的微型真菌将产生抗生素活性。来自该项目的数据将为第二阶段提供基础，在该阶段中，我们将继续开发我们新发现的先导抗生素。本项目的具体目标：目的1.为了评价真菌捕捉法培养微真菌的效果，我们用真菌捕捉器分离了200个非多余的真菌分离物，用标准平板法分离了50个真菌分离物。我们将比较来自真菌捕捉器和培养板的分离物的18S rDNA序列。目的2.为了鉴定抗生素活性，我们将进行真菌诱捕器来源的真菌分离株产生抗生素的中试筛选：我们将筛选200个真菌诱捕器来源的真菌分离株的抗生素活性，并优先选择50个有效的产生抗生素的菌株进行进一步分析。目的3.通过对25株真菌诱捕剂来源的高优先级菌株的活性成分进行分析，并将其与已知抗生素进行比较，以确定新的先导抗生素。

DESCRIPTION (provided by applicant): The long term goal of this project, is to develop novel antibiotics active against clinically important multi-drug- resistant (MDR) bacterial pathogens such as methicillin resistant Staphylococcus aureus (MRSA), and highly drug resistant Gram negative pathogens like Pseudomonas auruginosa. The goal of obtaining novel antibiotics will be achieved by employing a newly developed method (the 'fungal trap") to isolate and grow previously uncultured microfungi as a source of new antibiotic molecules. The majority of antibiotics in use today were discovered from environmental microorganisms, including microfungi. However, known microbe-derived antibiotics were discovered from only a very small fraction (<1%) of the diverse microorganisms that are culturable using standard techniques. Microfungi are a source of numerous valuable drugs used to treat diseases such as, bacterial infections (e.g. cephalosporins), fungal infections (e.g. candins), hypercholesterolemia (statins), and inflammatory diseases (e.g. cyclosporin). The world-wide market for antibiotics exceeds $20 billion. Sorely needed antibiotics active against MDR pathogens have the potential to yield a substantial market opportunity. In this Phase I project, we will evaluate and implement the "fungal-trap" method for isolating difficult to culture environmental microfungi, and we will generate a proof-of-principle that microfungi cultured using this method will produce antibiotic activities. Data derived from this project will provide the basis for phase II in which we will continue to develop our newly discovered lead antibiotics. Specific aims of the project: Aim 1. To evaluate the fungal trap method for culturing microfungi, we will isolate 200 nonredundant fungal isolates using fungal traps and 50 fungal isolates using standard plating methods. We will compare 18s rDNA sequences of isolates derived from fungal traps and Petri plates. Aim 2. To identify antibiotic activities, we will perform a pilot screen for antibiotic production by fungal isolates derived from fungal traps: we will screen 200 fungal trap derived isolates for antibiotic activity, and prioritize 50 validated antibiotic-producing isolates for further analysis. Aim 3. To identify novel lead antibiotics, we will analyze the active components from 25 high priority fungal- trap derived isolates and compare them against known antibiotics.