AHL Hydrolase Mediated Biofilm Inhibition

AHL 水解酶介导的生物膜抑制

• 批准号: 7538891
• 负责人: Robert J. Turner
• 金额: $ 13.3万
• 依托单位: LYBRADYN, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7538891
• 关键词: Admission activity; American Type Culture Collection; Antibiotic Resistance; Antibiotics; Apoenzymes; Bacteria; Bacterial Infections; Binding; Chemical Warfare Agents; Class; Coculture Techniques; Complex; Development; Devices; Drug Formulations; Effectiveness; Elements; Engineering; Enzymatic Biochemistry; Enzymes; Genetic Engineering; Gluconolactonase; Gram-Negative Bacteria; Growth; Homologous Gene; Hospitals; Host Defense Mechanism; Hydrolase; Hydrolysis; Implant; Infection; Kinetics; Laboratory Research; Lactones; Lead; Letters; Life; Link; Liquid substance; Literature; Marines; Marketing; Mediating; Medical Device; Microbial Biofilms; Molecular; N-(3-oxododecanoyl)homoserine lactone; N-butyrylhomoserine lactone; Nosocomial Infections; Numbers; Organism; Patients; Pesticides; Phase; Phosphoric Triester Hydrolases; Prevention; Process; Production; Pseudomonas; Pseudomonas aeruginosa; Public Health; Range; Rate; Relative (related person); Rhodococcus; Series; Signal Transduction; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Solubility; Structure; Surface; System; Testing; Time; Virulence Factors; Work; amidase; base; clinically relevant; commercialization; concept; directed evolution; enzyme structure; homoserine lactone; implantation; nosocomial infection control; novel; pathogen; physical property; prevent; quorum sensing; research and development; thermostability

DESCRIPTION (provided by applicant): Bacterial biofilms thwart the control of nosocomial infections, such as those resulting from medical device implantation. Elevated levels of one or more quorum sensing (QS) signals have been linked to the formation of bacterial biofilms, which makes eradication of bacteria colonizing the growth-amenable surfaces of implants difficult due to their biofilm-mediated resistance to antibiotics and host defense mechanisms. Therefore, it is desirable to develop ways to control quorum sensing and thus biofilm formation of P. aeruginosa and other Gram-negative pathogens. As with many Gram-negative organisms, N-acyl homoserine lactones (AHL) function as major QS signals for P. aeruginosa, specifically N-butanoyl-l-homoserine lactone (C(4)-HSL) and N-(3-oxododecanoyl)-l-homoserine lactone (3-oxo-C(12)-HSL). A small number of enzymes have been found that show potential for modulating QS by degrading AHLs, but there development has been limited. We have isolated enzymes from Geobacilli that possess demonstrable activity toward AHLs (functioning as lactonases) and are able to disrupt AHL-dependent processes of test organisms in liquid culture, including P. aeruginosa biofilm formation. Our most active Geobacillus enzyme, a very stable, soluble enzyme that can be readily purified, is already being expressed in a commercially viable system. We propose to further characterize these enzymes, relative to known AHL lactonases, to judge their suitability for our ultimate objective to formulate these enzymes into coatings for medical device products. PUBLIC HEALTH RELEVANCE Preventing medical device-related Pseudomonas aeruginosa (PA) infections by inhibiting quorum sensing (QS) dependent processes such as biofilm formation, allowing host defense mechanisms and antibiotic strategies to be more effective, is a concept that holds great potential. Towards these ends, we discovered enzymes isolated from species of Geobacillus are capable of hydrolyzing the lactone ring of N-acyl-L- homoserine lactones, the molecular trigger of bacterial QS for Gram-negative bacteria. Furthermore, co-culturing purified these Geobacillus enzymes inhibits QS-dependent processes, such as PA biofilm formation. In Phase I, we will thoroughly characterize the lactonase activities of our Geobacillus enzymes. We will also quantitatively compare our Geobacillus enzymes to other emerging lactonase enzymes.

描述（由申请人提供）：细菌生物膜阻止对医院感染的控制，例如由医疗装置植入引起的感染。一种或多种群体传感（QS）信号的水平升高与细菌生物膜的形成有关，这使得由于其生物膜介导的对抗生素和宿主防御机制的耐药性而难以植入植入物的生长不融合表面。因此，希望开发控制群体感应的方法，从而形成铜绿假单胞菌和其他革兰氏阴性病原体。与许多革兰氏阴性生物一样，N-酰基均同氨酸内酯（AHL）充当铜绿假单胞菌的主要QS信号，特别是N-叔丁酰基l-l-Homoserine lactone（C（4）-HSL）和N-（3-氧化二烷酰基）-L-HOMOSERINE-LACTONE（3-of-homoserine-lactone（3-of）-COxo-c（3-oxo-coxo-c）（12-oxco）（12）。已经发现少数酶通过降解AHL显示了调节QS的潜力，但其发育却有限。我们从地球杆菌中具有分离的酶，这些酶具有对AHL的可观活性（充当乳糖酶），并且能够破坏液态培养物中测试生物的AHL依赖性过程，包括铜绿假单胞菌生物膜形成。我们最活跃的地理酶酶是一种非常稳定的可溶酶，可以很容易纯化，已经在商业上可行的系统中表达。我们建议进一步将这些酶相对于已知的AHL lactonases，以判断它们适合我们将这些酶制成医疗设备产品涂料的最终目标。通过抑制诸如生物膜形成之类的依赖性过程（QS）依赖性过程，允许宿主的防御机制和抗生素策略更有效，可以更有效地使用公共卫生相关性，以防止医疗设备相关的铜绿假单胞菌（PA）感染。在这些末端，我们发现从地球杆菌物种中分离出来的酶能够水解N-酰基-l- hom- hom- hom- hom- hom- homeerine内酯的内酯环，这是革兰氏阴性细菌的细菌QS的分子触发。此外，共培养纯化这些地理酶抑制了QS依赖性过程，例如PA生物膜形成。在第一阶段，我们将彻底表征我们地理酶酶的乳酸酶活性。我们还将定量地将我们的地理酶酶与其他新兴的乳酸酶酶进行比较。