| 项目编号: | 1403748
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| 项目名称: | Collaborative Research: Creating a conductive connection between redox enzymes |
| 作者: | Ronald Koder
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| 承担单位: | CUNY City College
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| 批准年: | 2013
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| 开始日期: | 2014-07-01
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| 结束日期: | 2017-06-30
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| 资助金额: | USD300000
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| 资助来源: | US-NSF
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| 项目类别: | Standard Grant
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| 国家: | US
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| 语种: | 英语
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| 特色学科分类: | Engineering - Chemical, Bioengineering, Environmental, and Transport Systems
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| 英文关键词: | enzyme
; redox center
; activity
; redox enzyme
; collaborative research
; wiring redox protein
; single ping pong enzyme
; cofactor-dependent enzyme
; known redox enzyme
; ping pong redox enzyme
; unrelated redox protein
; wiring other redox protein
; final enzyme
; collaborative project
; enzyme active site
; redox protein
; additional enzyme
; new enzyme complex
|
| 英文摘要: | PI's: Koder, Ronald / Banta, Scott A.
Proposal Numbers: 1403748 / 1402913
Institutions: CUNY City College / Columbia University
Title: Collaborative Research: Simplifying metabolic pathways by wiring redox proteins together
Energy transfer between enzymatic proteins is often accomplished using small molecule cofactors, such as NAD(H) and NADP(H). This system is advantageous in biological systems; however, in technology applications the use of these cofactors is undesirable for several reasons including their high cost and low stability. Advances in protein design and protein engineering have enabled tremendous advances in synthetic biology where new proteins with novel, unnatural functions can be created and characterized. In this project a new protein will be made by coupling two cofactor-dependent enzymes together. In order to eliminate the need for the cofactor, the enzymes will be connected with a designed 'staple' peptide system that contains redox center that enable electron transfer between the enzymes. This will 'wire' the proteins together so that neither one requires the unstable cofactor molecules for activity. Combined, this new enzyme complex will have an activity that is not found in nature. And this will chart a path forward for wiring other redox proteins together towards a goal of eliminating cofactor requirements in future synthetic biology projects. The team consists of PIs from City College of New York and Columbia University. There will be several outreach activities associated with this project including interactions with local high school teachers and students.
The overall goal of this collaborative project is to use protein engineering to wire two unrelated redox proteins together creating a novel cofactor-less enzymatic reaction. The investigators will attempt to demonstrate the first example of the wiring of two redox proteins together to form a ping pong redox enzyme with an activity not found in nature. They will start with two known redox enzymes: formate dehydrogenase and lactate dehydrogenase. These enzymes will be connected by a novel 'collagen staple' system that will enable specific self-assembly and the introduction of redox centers for direct electron transfer between the two enzyme active sites. The final enzyme will have a completely novel function: pyruvate reduction to lactate with the concomitant oxidation of formate to carbon dioxide. The new kinetic mechanism and activity of the resultant biocatalyst will be extensively characterized. This approach can then be extended to combine additional enzymes, creating new cofactor-less biocatalysts for industrially important oxidation and reduction reactions. This approach will allow to combine redox enzymes with sequential kinetic mechanisms into a single ping pong enzyme with a synthetic redox center. The research is integrated with educational activities which introduces undergraduates, graduate students and New York City Public High School teachers to the interdisciplinary science of biophysics. These efforts will encourage young students to pursue careers in Science, Technology, Engineering and Mathematics.
This award by the Biotechnology, Biochemical, and Biomass Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology. |
| 资源类型: | 项目
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| 标识符: | http://119.78.100.158/handle/2HF3EXSE/96429
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| Appears in Collections: | 影响、适应和脆弱性
气候减缓与适应
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| Recommended Citation: | |
Ronald Koder. Collaborative Research: Creating a conductive connection between redox enzymes. 2013-01-01.
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