期刊
METABOLIC ENGINEERING
卷 33, 期 -, 页码 41-51出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2015.10.006
关键词
Pathway engineering; High-throughput screening; Biosensor; Deoxyviolacein biosynthesis; Sequential optimization
资金
- National Natural Science Foundation of China [NSFC 21376137]
- Tsinghua University Initiative Scientific Research Program [2013Z02-1]
- Ministry of Science and Technology of China (973) [2013CB734001]
- CAS Interdisciplinary Innovation Team [Y429012CX8]
Because high-throughput screening tools are typically unavailable when using the pathway-engineering approach, we developed a new strategy, named intermediate sensor-assisted push-pull strategy, which enables sequential pathway optimization by incorporating a biosensor targeting a key pathway intermediate. As proof of concept, we constructed an L-Trp biosensor and used it to optimize the deoxyviolacein biosynthetic pathway, which we divided into two modules with L-Trp being the product of the upstream and the substrate of the downstream module for deoxyviolacein synthesis. Using the biosensor and fluorescence-activated cell sorting, the activities of the two modules were sequentially and independently optimized in Escherichia coil to achieve the desired phenotypes. By this means, we increased the deoxyviolacein titer 4.4-fold (1.92 g/L), which represents the greatest deoxyviolacein production reported. This work suggests that a biosynthetic pathway can be enhanced to produce a value-added secondary metabolite(s) without available end-product screening method by using a central metabolic junction molecule biosensor(s). (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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