Isolating novel fungi for antibiotic discovery

描述（由申请人提供）：本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（如耐甲氧西林金黄色葡萄球菌（MRSA））和高耐药革兰氏阴性病原体（如金黄色假单胞菌）具有活性的新型抗生素。获得新抗生素的目标将通过采用新开发的方法（“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来实现。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，已知的微生物衍生的抗生素仅从使用标准技术可培养的多种微生物中的非常小的一部分（<1%）中发现。微真菌是用于治疗疾病的许多有价值的药物的来源，所述疾病例如细菌感染（例如头孢菌素）、真菌感染（例如念珠菌）、高胆固醇血症（他汀类）和炎性疾病（例如环孢菌素）。全球抗生素市场超过200亿美元。迫切需要的对MDR病原体有活性的抗生素有可能产生巨大的市场机会。在第一期项目中，我们将评估和实施用于分离难以培养的环境微真菌的“真菌陷阱”方法，并将产生使用该方法培养的微真菌将产生抗生素活性的原理证明。从该项目获得的数据将为第二阶段提供基础，在第二阶段，我们将继续开发我们新发现的先导抗生素。项目的具体目标：目标1。为了评估培养微真菌的真菌陷阱方法，我们将使用真菌陷阱分离200个非冗余真菌分离株，使用标准平板法分离50个真菌分离株。我们将比较来自真菌陷阱和培养皿的分离物的18S rDNA序列。目标2.为了鉴定抗生素活性，我们将通过来自真菌陷阱的真菌分离株进行抗生素生产的中试筛选：我们将筛选200种来自真菌陷阱的分离株的抗生素活性，并优先考虑50种经验证的产抗生素分离株进行进一步分析。目标3：为了鉴定新的先导抗生素，我们将分析来自25种高优先级真菌诱捕器衍生的分离物的活性组分，并将它们与已知的抗生素进行比较。

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.