Production and application of recombinant biopolymers with exotic chemistries

具有外来化学成分的重组生物聚合物的生产和应用

基本信息

批准号：
9109978
负责人：
Miriam Amiram
金额：
$ 5.05万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9109978
关键词：
Address Adhesives Adverse effects Albumins Alkynes Amino Acids Amino Acyl-tRNA Synthetases Anti-Bacterial Agents Antibiotic Resistance Award Azides Back Bacteria Bacterial Drug Resistance Binding Biocompatible Materials Biological Process Biology Biopolymers Bypass Cell Differentiation process Chemicals Chemistry Codon Nucleotides Communication Complex Cues Data Development Differentiation and Growth Disease Dopa Drug Delivery Systems Drug Kinetics Elastin Engineering Escherichia coli Eukaryotic Cell Evolution Family Fatty Acids Formulation Frequencies Genome engineering Goals Half-Life Hybrids Injection of therapeutic agent Insulin Libraries Life Ligation Magnetism Mediating Mentors Metals Methanococcus Methanosarcina Methodology Modeling Multi-Drug Resistance Nature Organism Peptides Pharmacologic Substance Phase Phase Transition Phenylalanine Polymers Positioning Attribute Post-Translational Protein Processing Production Property Proteins Recombinants Research Research Training Resistance Science Sense Codon Silver Site System Technology Terminator Codon Therapeutic Agents Tissues Training Transfer RNA Translating Translations Tyrosine Universities Variant Vision Work adhesive protein (mussel)antimicrobial antimicrobial drug antimicrobial peptide base biological systems biomaterial compatibility career development chemical group clinically relevant combat cycloaddition design drug resistant bacteria experience functional group glucagon-like peptide glucagon-like peptide 1 improved in vivo nanoparticle novel pathogen peptide drug polypeptide public health relevance pyrrolysine responsible research conduct scaffold skills synthetic biology

Address Adhesives Adverse effects Albumins Alkynes Amino Acids Amino Acyl-tRNA Synthetases Anti-Bacterial Agents Antibiotic Resistance Award Azides Back Bacteria Bacterial Drug Resistance Binding Biocompatible Materials Biological Process Biology Biopolymers Bypass Cell Differentiation process Chemicals Chemistry Codon Nucleotides Communication Complex Cues Data Development Differentiation and Growth Disease Dopa Drug Delivery Systems Drug Kinetics Elastin Engineering Escherichia coli Eukaryotic Cell Evolution Family Fatty Acids Formulation Frequencies Genome engineering Goals Half-Life Hybrids Injection of therapeutic agent Insulin Libraries Life Ligation Magnetism Mediating Mentors Metals Methanococcus Methanosarcina Methodology Modeling Multi-Drug Resistance Nature Organism Peptides Pharmacologic Substance Phase Phase Transition Phenylalanine Polymers Positioning Attribute Post-Translational Protein Processing Production Property Proteins Recombinants Research Research Training Resistance Science Sense Codon Silver Site System Technology Terminator Codon Therapeutic Agents Tissues Training Transfer RNA Translating Translations Tyrosine Universities Variant Vision Work adhesive protein (mussel)antimicrobial antimicrobial drug antimicrobial peptide base biological systems biomaterial compatibility career development chemical group clinically relevant combat cycloaddition design drug resistant bacteria experience functional group glucagon-like peptide glucagon-like peptide 1 improved in vivo nanoparticle novel pathogen peptide drug polypeptide public health relevance pyrrolysine responsible research conduct scaffold skills synthetic biology

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项目摘要

DESCRIPTION (provided by applicant): Increasingly demanding biomedical and biotechnological endeavors require sophisticated materials that can respond to multiple environmental cues, interact with biological systems, template compound assemblies, and direct the growth and differentiation of cells and tissues. My long term research goal is to create customized and functional protein-based biomaterials containing genetically encoded non- standard amino acids (nsAAs) to site-specifically introduce diverse chemical functionalities. Towards this research vision, my research objectives are two-fold. First, I will work to expand and accelerate our ability to produce protein polymers containing multiple nsAAs. To this end I will develop a novel evolution platform and extensive mutagenized libraries for components used to incorporate nsAAs. Second, I will utilize this platform and variants isolated to introduce novel functions and properties to protein-based biomaterials with the goal of developing antimicrobial and drug delivery formulations. This proposal is designed leverage Prof. Isaacs' expertise in synthetic biology, Prof. Soll's expertise in systems for nsAA incorporation, Prof. Saltzman's expertise in biomaterial-based drug delivery, and Dr. Amiram's previous research experience in protein-polymer synthesis, design and application. In addition to scientific training several career development activities will be pursued during this award including attending career development courses offered by Yale University in science and communication skills, developing mentoring skills, and obtaining extensive training in responsible conduct of research. In our preliminary data, we detail a novel evolution platform for aminoacyl-tRNA synthetases (AARS) for nsAA incorporation. We establish that using evolved AARS variants expressed in a genomically recoded E. coli, we are able to bypass former limitations of nsAA incorporation and achieve high yield and high purity production of biopolymers containing multiple (10-30) nsAA instances. In Aim 1, I will extend and expand AARS libraries to enable evolution of translation components for the incorporation adhesive and bulky nsAAs, which can also be expressed and utilized in prokaryotic and eukaryotic organisms. In Aim 2, I will incorporate multiple azide groups to mediate bioorthogonal attachment of multiple fatty acid molecules to improve peptide and protein pharmacokinetics by enhanced albumin binding. In Aim 3, I will generate adhesive DOPA-biomaterials to create silver-containing formulations for antibacterial treatment. Ultimately, I anticipate that my work will facilitate a fundamental change in our abilities to desin and impart new functions and properties to protein- based materials. The results of these studies will yield enabling technologies and strains for multi-site nsAA incorporation, new classes of biopolymers, customized materials with novel functions, and design rules for manipulating and tuning biopolymer properties.

描述（由适用提供）：越来越多的生物医学和生物技术努力需要复杂的材料，这些材料可以响应多种环境线索，与生物系统，模板化合物组件相互作用，并指导细胞和组织的生长和分化。我的长期研究目标是创建定制且功能性蛋白质的生物材料，其中包含一般编码的非标准氨基酸（NSAAS），以特定于现场引入潜水化学功能。针对这一研究愿景，我的研究目标是两个方面。首先，我将努力扩展和加速我们生产含有多种NSAA的蛋白质聚合物的能力。为此，我将开发一个新颖的进化平台，并为用于合并NSAAS的组件进行广泛的诱变库。其次，我将利用这个平台，并分离出来，将新型功能和特性引入基于蛋白质的生物材料，以开发抗菌和药物递送配方。该提案旨在设计杠杆ISAACS教授在合成生物学方面的专业知识，SOLL教授在NSAA Corporation的系统专业知识，Saltzman教授在基于生物材料的药物递送方面的专业知识以及Amiram博士先前在蛋白质 - 聚合物合成，设计和应用方面的研究经验。除科学培训外，还将在本奖项中进行几项职业发展活动，包括参加耶鲁大学科学和沟通技巧提供的职业发展课程，发展指导技能以及在负责任的研究中获得广泛的培训。在我们的初步数据中，我们详细介绍了NSAA Corporation氨基酰基-TRNA合成酶（AARS）的新型演化平台。我们确定使用在基因组回收的大肠杆菌中表达的进化的AARS变体，我们在AIM 2中，我将结合多个叠氮化物组，以通过增强的专辑结合来介导多脂肪酸分子的生物句子附着，以改善肽和蛋白质药代动力学。在AIM 3中，我将生成粘合剂DOPA-BIOMATERIANS，以创建含白银的配方进行抗菌治疗。最终，我预计我的工作将支持我们的能力的根本变化，并将新功能和特性赋予基于蛋白质的材料。这些研究的结果将为多站点NSAA成立，新的生物聚合物，具有新功能的定制材料以及操纵和调整生物聚合物特性的设计规则而产生促成技术和菌株。