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dc.contributor.author
Barstow, Buz
dc.contributor.author
Agapakis, Christina M.
dc.contributor.author
Boyle, Patrick M.
dc.contributor.author
Grandl, Gerald
dc.contributor.author
Silver, Pamela A.
dc.contributor.author
Wintermute, Edwin H.
dc.date.accessioned
2019-10-03T13:18:18Z
dc.date.available
2017-06-11T16:51:38Z
dc.date.available
2019-10-03T13:18:18Z
dc.date.issued
2011
dc.identifier.issn
1754-1611
dc.identifier.other
10.1186/1754-1611-5-7
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/99758
dc.identifier.doi
10.3929/ethz-b-000099758
dc.description.abstract
Background FeFe-hydrogenases are the most active class of H2-producing enzymes known in nature and may have important applications in clean H2 energy production. Many potential uses are currently complicated by a crucial weakness: the active sites of all known FeFe-hydrogenases are irreversibly inactivated by O2. Results We have developed a synthetic metabolic pathway in E. coli that links FeFe-hydrogenase activity to the production of the essential amino acid cysteine. Our design includes a complementary host strain whose endogenous redox pool is insulated from the synthetic metabolic pathway. Host viability on a selective medium requires hydrogenase expression, and moderate O2 levels eliminate growth. This pathway forms the basis for a genetic selection for O2 tolerance. Genetically selected hydrogenases did not show improved stability in O2 and in many cases had lost H2 production activity. The isolated mutations cluster significantly on charged surface residues, suggesting the evolution of binding surfaces that may accelerate hydrogenase electron transfer. Conclusions Rational design can optimize a fully heterologous three-component pathway to provide an essential metabolic flux while remaining insulated from the endogenous redox pool. We have developed a number of convenient in vivo assays to aid in the engineering of synthetic H2 metabolism. Our results also indicate a H2-independent redox activity in three different FeFe-hydrogenases, with implications for the future directed evolution of H2-activating catalysts.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
BioMed Central
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/2.0/
dc.title
A synthetic system links FeFe-hydrogenases to essential E. coli sulfur metabolism
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 2.0 Generic
dc.date.published
2011-05-26
ethz.journal.title
Journal of Biological Engineering
ethz.journal.issue
5
en_US
ethz.journal.abbreviated
J. biol. eng.
ethz.pages.start
7
en_US
ethz.size
15 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.nebis
005513341
ethz.publication.place
London, UK
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02070 - Dep. Gesundheitswiss. und Technologie / Dep. of Health Sciences and Technology::02701 - Inst.f. Lebensmittelwiss.,Ernährung,Ges. / Institute of Food, Nutrition, and Health::03819 - Wolfrum, Christian / Wolfrum, Christian
en_US
ethz.leitzahl.certified
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02070 - Dep. Gesundheitswiss. und Technologie / Dep. of Health Sciences and Technology::02701 - Inst.f. Lebensmittelwiss.,Ernährung,Ges. / Institute of Food, Nutrition, and Health::03819 - Wolfrum, Christian / Wolfrum, Christian
ethz.date.deposited
2017-06-11T16:52:04Z
ethz.source
ECIT
ethz.identifier.importid
imp593653125b44576594
ethz.ecitpid
pub:156078
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-07-13T09:12:48Z
ethz.rosetta.lastUpdated
2019-10-03T13:18:32Z
ethz.rosetta.versionExported
true
ethz.COinS
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