Isolating novel fungi for antibiotic discovery

项目描述（由申请人提供）：该项目的长期目标是开发具有抗临床重要多重耐药（MDR）细菌病原体（如耐甲氧西林金黄色葡萄球菌（MRSA））和高度耐药革兰氏阴性病原体（如金毛假单胞菌）活性的新型抗生素。获得新型抗生素的目标将通过采用一种新开发的方法（“真菌陷阱”）来分离和培养以前未培养的微真菌作为新抗生素分子的来源。目前使用的大多数抗生素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仅从可使用标准技术培养的多种微生物的极小部分（<1%）中发现。微真菌是用于治疗诸如细菌感染（如头孢菌素）、真菌感染（如念珠菌素）、高胆固醇血症（他汀类药物）和炎症性疾病（如环孢素）等疾病的许多有价值药物的来源。全球抗生素市场超过200亿美元。对耐多药耐药性病原体具有活性的急需抗生素有可能产生巨大的市场机会。在本一期项目中，我们将评估和实施“真菌陷阱”方法，用于分离难以培养的环境微真菌，我们将产生用这种方法培养的微真菌产生抗生素活性的原理证明。从这个项目中获得的数据将为第二阶段提供基础，我们将继续开发我们新发现的含铅抗生素。项目的具体目标：目标1。为了评估真菌陷阱培养微真菌的方法，我们将用真菌陷阱分离200株非冗余真菌，用标准电镀方法分离50株真菌。我们将比较从真菌陷阱和培养皿中分离的18s rDNA序列。目标2。为了确定抗生素活性，我们将对来自真菌陷阱的真菌分离株进行抗生素生产的试点筛选：我们将筛选200个真菌陷阱衍生的分离株的抗生素活性，并优先考虑50个经过验证的产生抗生素的分离株进行进一步分析。目标3。为了鉴定出新的先导抗生素，我们将分析25种高优先级真菌诱捕器分离物的活性成分，并将其与已知抗生素进行比较。

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.