| 项目编号: | 1605465
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| 项目名称: | SusChEM: Development of a Protecting Group Toolkit for Metabolic Engineering |
| 作者: | John Dueber
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| 承担单位: | University of California-Berkeley
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| 批准年: | 2016
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| 开始日期: | 2016-07-01
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| 结束日期: | 2019-06-30
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| 资助金额: | 350020
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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: | group
; control
; toolkit
; functional group reactivity
; engineering metabolic pathway
; rational engineering
; cell
; natural chemical group
; metabolic engineering application
; engineering community
; group strategy
; chemical
; john e. engineering living cell
; biochemical engineering program
|
| 英文摘要: | 1605465
Dueber, John E.
Engineering living cells to perform as chemical factories offers a route for sustainable production of a variety of chemicals. Microbial cells can rapidly self-replicate on inexpensive substrates and make enzymes capable of catalyzing difficult chemical reactions cheaply, quickly, and cleanly. However, the cell presents a complex environment for conducting these multi-step syntheses and, accordingly, undesired reactivity often occurs to produce undesired chemical products or, even worse, result in toxicity to the microbial production host. Multi-step organic chemical syntheses conducted in test tubes often employ chemical protecting groups to gain control over when and where sites of a chemical substrate will be reactive. This general approach will be mimicked in the cell by employing natural chemical groups that can be reversibly added to achieve similar control over reactivity of a chemical in the cell. Microbial strains capable of catalyzing efficient protection and stability of the resulting tailored products will be of broad interest to metabolic engineers and will be made available to the scientific and engineering communities.
Engineering metabolic pathways to sustainably produce molecules of interest requires designed control at multiple levels. Numerous problems can limit productivity, including metabolite toxicity, off-pathway catalysis, and failure to secrete the final product. Various synthetic biology approaches have been developed for introducing control at the DNA, RNA, and protein levels to address many of these challenges. In this proposal, control at the metabolite level is targeted for an additional strategy in this toolkit. The protecting group strategy used in synthetic organic chemistry to gain control over functional group reactivity will be mimicked to gain similar control over when and where a molecule is active. Three biomolecular tailoring groups (glucosyl, acetyl, and sulfonyl) are proposed to provide reversible protection in addition to the ability to tailor for altered properties such as solubility and membrane permeability as well as the recognition by other enzymes that could lower toxicity. Thus, reactive molecules can be made chemically inert while protected and then reactivity be reinstated when desired via enzymatic deprotection. Enabling the use of the appropriate protecting group for the desired application demands the construction of a toolkit of strains wherein each of these protected products will be stable. For many of these protecting groups, several enzymes will need to be knocked out of the production strain to ensure small molecule product stability in the cellular environment. A high-throughput colorimetric plate assay will be employed to screen large matrices of gene target knockouts. Furthermore, it is critical that these protecting groups do not limit product titers or production rates. Accordingly, rational engineering and adaptations for increased flux for each protection reaction will be performed. Although acetylation and glucosylation are expected to already have fairly high capacity, sulfonation capacity is extremely low. The resultant strains should prove broadly useful for a variety of metabolic engineering applications and will accordingly be shared with the community.
This award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biosciences. |
| 资源类型: | 项目
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| 标识符: | http://119.78.100.158/handle/2HF3EXSE/92011
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| Appears in Collections: | 全球变化的国际研究计划
科学计划与规划
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| Recommended Citation: | |
John Dueber. SusChEM: Development of a Protecting Group Toolkit for Metabolic Engineering. 2016-01-01.
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