Journal
JOURNAL OF BIOTECHNOLOGY
Volume 192, Issue -, Pages 192-196Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jbiotec.2014.10.015
Keywords
Cohesin-dockerin interaction; Saccharomyces cerevisiae; Substrate channeling; Synthetic scaffold; 2,3-Butanediol
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Funding
- National Research Foundationof Korea (NRF) Grant - Korean Government [2012R1A1A-3011963]
- National Research Foundation of Korea [2012R1A1A3011963] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Substrate channeling is a process of transferring an intermediate from one enzyme to the next enzyme without diffusion into the bulk phase, thereby leading to an enhanced reaction rate. Here, we newly designed substrate channeling modules in Saccharomyces cerevisiae based on a high affinity interaction between dockerin and cohesin domains, which is a key process in the formation of cellulosome structure. Synthetic scaffolds containing two, three, or seven cohesin domains were constructed, and the assembly of dockerin-tagged proteins onto the scaffolds was confirmed by pull-down assay and bimolecular fluorescent complementation (BiFC) assay in vivo. This system was applied to produce 2,3-butanediol in S. cerevisiae by using dockerin-tagged AlsS, AlsD, and Bdh1 enzymes, resulting in a gradual increase in 2,3-butanediol production depending on the number of cohesin domains in the scaffold. (C) 2014 Elsevier B.V. All rights reserved.
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