Synthesis and properties of a bacterial bioadhesive

细菌生物粘附剂的合成及性能

• 批准号: 8344340
• 负责人: YVES V BRUN
• 金额: $ 39.67万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8344340
• 关键词: Adherence; Adhesions; Adhesiveness; Adhesives; Age; Aging; Antitoxins; Apoptosis; Area; Atomic Force Microscopy; Bacteria; Bacterial Infections; Base Sequence; Binding; Biochemical; Biocompatible Materials; Biological; Biological Process; Biophysics; Caulobacter; Caulobacter crescentus; Cell Death; Cells; Chemicals; DNA; DNA Binding; DNA Sequence; Data; Deacetylase; Deacetylation; Dental; Development; Employee Strikes; Environment; Exhibits; Fluorescence Microscopy; Fresh Water; Future; Genes; Genetic; Goals; Human Activities; Human body; Image; Infection; Ionic Strengths; Kinetics; Marines; Measures; Mechanics; Medical; Methods; Microbial Biofilms; Mutation; Operative Surgical Procedures; Oxygen; Polysaccharides; Production; Property; Research; Resolution; Role; Saline; Solutions; Solvents; Specificity; Spectrum Analysis; Structure; Surface; System; Testing; Toxin; base; crosslink; design; extracellular; improved; inhibitor/antagonist; insight; interdisciplinary approach; mutant; pathogen; physical property; polysaccharide deacetylase; research study; response; shear stress

DESCRIPTION (provided by applicant): Bacteria often utilize polysaccharides as adhesive structures to attach to surfaces, to form biofilms, and to infect host cells. In addition, polysaccharides hold strong promise as biological adhesives in many areas of human activity, including as dental and surgical adhesives. The bacterium Caulobacter crescentus synthesizes a polysaccharide called the holdfast that exhibits and impressive adhesive force. Contrary to most commercial adhesives, holdfasts adhere tightly to a variety of surfaces in both freshwater and marine environments. Such a property is critical for medical applications in the human body. The general goal of this research is to use a multidisciplinary approach ranging from genetics to biophysics to study the chemical and biophysical basis for holdfast properties and to understand how holdfast properties are modulated by deacetylation and inhibition by extracellular DNA (eDNA). The project has three specific aims. The first aim is to determine the biophysical basis for holdfast adhesiveness. Atomic force microscopy (AFM) will be used to systematically study the influence of surface roughness, shear stress, surface composition, ionic strength, and pH on holdfast adhesion in order to provide a better understanding of the mechanism of holdfast adhesion and adhesion control. The second aim is to determine the role of deacetylation in holdfast anchoring and adhesive properties. A holdfast polysaccharide deacetylase mutant causes the release of non-adherent holdfast in solution. The composition and structure of the holdfast polysaccharide will be determined from normal and deacetylase mutant cells. AFM force spectroscopy and high-resolution fluorescence microscopy will be used to determine the role of deacetylation on holdfast adhesiveness and cohesiveness. Biochemical experiments will be used to study the role of deacetylation in anchoring the holdfast to the cell. Finally, similar studies of the holdfast of marine species will provide better biomaterials for potential applications in the saline environment of the human body. The third aim is to determine the biological basis for the recently discovered mechanism of eDNA inhibition of holdfast adherence. The role of a toxin-antitoxin system in the production of eDNA by programmed cell death will be studied and the basis for the sequence specificity of holdfast inhibition will be determined. AFM indentation studies and simultaneous AFM imaging and Raman scattering spectroscopy of holdfasts bound or not to eDNA will be used to determine how specific DNA alters the structure and structural properties of the holdfast. Results from the proposed studies will provide insight into the basic mechanisms for the impressive adhesive properties of the holdfast and modulation of these properties, paving the way for the future development of the holdfast as a biological adhesive. In addition, results of these studies will provide insights into the mechanism of polysaccharide adhesiveness in general, as well as for strategies to inhibit polysaccharide adhesion, for example during infection by pathogens. PUBLIC HEALTH RELEVANCE: Bacteria often utilize polysaccharides as adhesive structures to attach to surfaces, to form biofilms, and to infect host cells, and these polysaccharides hold strong promise as biological adhesives in many areas of human activity, including as dental and surgical adhesives. We will study one of the strongest known biological adhesives, the holdfast polysaccharide of Caulobacter crescentus by investigating the chemical and the biophysical basis for holdfast properties and the mechanism of action of a specific inhibitor of holdfast adhesiveness. These studies will provide data essential to the design of strategies to use polysaccharides in various applications, as well as strategies to inhibit polysaccharide adhesion during infection by bacterial pathogens.

描述（由申请人提供）：细菌经常利用多糖作为粘合结构，以附着在表面上，形成生物膜并感染宿主细胞。此外，在人类活动的许多领域，包括牙科和外科粘合剂，多糖作为生物粘合剂具有很大的希望。细菌caulobacter Crescentus合成了一种称为Holdfast的多糖，表现出了令人印象深刻的粘合力。与大多数商业粘合剂相反，Holdfasts紧密地粘附在淡水和海洋环境中的各种表面。这种特性对于人体的医疗应用至关重要。这项研究的一般目标是使用从遗传学到生物物理学的多学科方法来研究持有特性的化学和生物物理基础，并了解如何通过脱乙酰基化和通过细胞外DNA（EDNA）抑制持续性的性能来调节持有性能。该项目具有三个特定的目标。第一个目的是确定持有粘合剂的生物物理基础。原子力显微镜（AFM）将用于系统地研究表面粗糙度，剪切应力，表面组成，离子强度和pH值对持有快速粘附的影响，以便更好地了解持有的固定粘附和粘附控制机制。第二个目的是确定脱乙酰化在固定锚定和粘合特性中的作用。固定快速多糖脱乙酰基酶突变剂导致溶液中非粘附的持有释放。 Holdfast多糖的组成和结构将由正常和脱乙酰基酶突变细胞确定。 AFM力光谱和高分辨率荧光显微镜将用于确定脱乙酰化对持有的粘附性和凝聚力的作用。生化实验将用于研究脱乙酰化在将固定固定固定到细胞中的作用。最后，对海洋物种持有的类似研究将为人体盐环境中的潜在应用提供更好的生物材料。第三个目的是确定最近发现的EDNA抑制持续性机制的生物学基础。将研究毒素 - 抗毒素系统在通过程序性细胞死亡产生EDNA中的作用，并将确定Holdfast抑制的序列特异性的基础。 AFM凹痕研究和同时进行的AFM成像以及与EDNA结合或不与EDNA的持有的拉曼散射光谱法被用来确定特定的DNA如何改变Holdfast的结构和结构特性。拟议的研究的结果将洞悉持有和调节这些特性的令人印象深刻的粘合性特性的基本机制，从而为Holdfast作为生物粘合剂的未来发展铺平了道路。此外，这些研究的结果将为您提供见解 多糖粘附性的机制通常以及抑制多糖粘附的策略，例如在病原体感染期间。 公共卫生相关性：细菌经常利用多糖作为粘合结构，与表面相连，形成生物膜并感染宿主细胞，这些多糖在人类活动的许多领域（包括牙科和手术粘合剂）的生物学粘合剂具有很强的希望。我们将通过研究Holdfast性能的化学和生物物理基础以及Holdfast粘合剂的特定抑制剂的作用机理，研究最强的已知生物粘合剂之一，这是Caulobacter Crescentus的持有快速多糖。这些研究将为设计在各种应用中使用多糖的策略以及抑制细菌病原体感染过程中抑制多糖粘附的策略提供必不可少的数据。