Studies on Calicheamicin Biosynthesis and Resistance

加利车霉素生物合成及耐药性研究

基本信息

批准号：
7432499
负责人：
Jon Scott Thorson
金额：
$ 27.54万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-05-01 至 2010-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7432499
关键词：
Address Anabolism Anthracycline Antibiotics Anthracyclines Architecture Base Sequence Biological Biological Factors Biological Models Clinical Cloning Complex Cosmids DNA delivery Disruption Dynemicin Enzyme Gene Enzymes Facility Construction Funding Category Family Figs - dietary Foundations Funding Gene Expression Genes Genetic Genetic Techniques Genomics Goals Grant Homologous Gene In Vitro Knowledge Libraries Methods Micromonospora Modeling Modification Molecular Natural Product Drug Numbers Participant Phase Phase I Clinical Trials Polymerase Chain Reaction Principal Investigator Production Proteins Proteolysis Protocols documentation Resistance Sequence Analysis Shotgun Sequencing Time Vision combinatorial comparative directed evolution in vivo insight novel polyketide synthase programs reconstitution resistance mechanism size southern hybridization sugar thiosugar tool

项目摘要

DESCRIPTION (provided by applicant): Calicheamicin gamma1 from Micromonospora echinospora spp. calichensis is the most prominent of the 10- membered enediyne family with respect to its unprecedented molecular architecture, spectacular biological activity and clinical value. As such, calicheamicin is an excellent target for the study of natural product biosynthesis and self-resistance. The objective of the first phase of this study was to i) pursue the biosynthesis of the DNA-delivery component of calicheamicin (the aryltetrasaccharide, comprised of four uniquely functionalized sugars), ii) develop the genetic tools (transformation and gene disruption protocols) to address calicheamicin biosynthesis in Micromonospora and iii) investigate the mechanism(s) of calicheamicin self-resistance in Micromonospora. With these goals achieved and new tools/information in place, the second phase of this massive project will predominately focus upon expanding this program toward understanding and exploiting the complex biosynthesis of the enediyne core. While continuing our focus upon calicheamicin as a model for 10-membered enediyne biosynthesis, a second complimentary 10-membered enediyne model will be pursued (namely, dynemicin from Micromonospora chersina) selected for its unique architecture (an unprecedented fused enediyne-anthracycline), predominate biological activity, anticipated small gene locus size (excellent for production of dynemicin in 'genetically-friendly' heterologous hosts) and the opportunity for comparative genomics of the calicheamicin and dynemicin biosynthetic loci. The fundamental vision of this program remains constant - to present rational strategies from which to build a foundation of knowledge regarding 10-membered enediyne biosynthesis and self-resistance; the consequence of which will continue to provide pioneering discoveries in enzymatic transformation, tools for the rational biosynthetic modification of natural product drug leads, the potential for enediyne overproducing strains and possibly even an enediyne combinatorial biosynthesis program.

描述（由申请人提供）：Micromonospora echinospora spp的Calicheamicin Gamma1。 Calichensis是10个成员的Enediyne家族中最杰出的，就其前所未有的分子结构，壮观的生物学活性和临床价值而言。因此，钙甲氨基蛋白是研究天然产物生物合成和自我抗性的绝佳靶标。 The objective of the first phase of this study was to i) pursue the biosynthesis of the DNA-delivery component of calicheamicin (the aryltetrasaccharide, comprised of four uniquely functionalized sugars), ii) develop the genetic tools (transformation and gene disruption protocols) to address calicheamicin biosynthesis in Micromonospora and iii) investigate the mechanism(s) of Calicheamicin在微孔孢子中的自我抗性。随着实现这些目标和新的工具/信息，这个大规模项目的第二阶段将主要集中于将该计划扩展到理解和利用Enediyne核心的复杂生物合成。 While continuing our focus upon calicheamicin as a model for 10-membered enediyne biosynthesis, a second complimentary 10-membered enediyne model will be pursued (namely, dynemicin from Micromonospora chersina) selected for its unique architecture (an unprecedented fused enediyne-anthracycline), predominate biological activity, anticipated small gene locus size (excellent for production of在“基因友好型”异源宿主中的Dynemicin和Calicheamicin和Dynemicin Biosynthetic locynthetic loci的比较基因组学的机会。该计划的基本愿景仍然持续不变 - 提出理性策略，从中建立了关于10元的Enediyne生物合成和自我抗性的知识的基础；其后果将继续提供酶促转化的开创性发现，自然产物药物的理性生物合成修饰的工具，可能导致过度产生菌株，甚至可能是Enediynenatorial Compinatorial生物合成计划。