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Elimination of sucr...
Elimination of sucrose transport and hydrolysis in Saccharomyces cerevisiae : a platform strain for engineering sucrose metabolism
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Marques, Wesley Leoricy (författare)
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Mans, Robert (författare)
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Marella, Eko Roy (författare)
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Cordeiro, Rosa Lorizolla (författare)
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van den Broek, Marcel (författare)
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Daran, Jean-Marc G. (författare)
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Pronk, Jack T. (författare)
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Gombert, Andreas K. (författare)
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- van Maris, Antonius J. A. (författare)
- KTH,Industriell bioteknologi
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(creator_code:org_t)
- 2017-01-13
- 2017
- Engelska.
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Ingår i: FEMS yeast research (Print). - : Oxford University Press. - 1567-1356 .- 1567-1364. ; 17:1
- Relaterad länk:
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https://doi.org/10.1...
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https://academic.oup...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Many relevant options to improve efficacy and kinetics of sucrose metabolism in Saccharomyces cerevisiae and, thereby, the economics of sucrose-based processes remain to be investigated. An essential first step is to identify all native sucrose-hydrolysing enzymes and sucrose transporters in this yeast, including those that can be activated by suppressor mutations in sucrose-negative strains. A strain in which all known sucrose-transporter genes (MAL11, MAL21, MAL31, MPH2, MPH3) were deleted did not grow on sucrose after 2 months of incubation. In contrast, a strain with deletions in genes encoding sucrose-hydrolysing enzymes (SUC2, MAL12, MAL22, MAL32) still grew on sucrose. Its specific growth rate increased from 0.08 to 0.25 h(-1) after sequential batch cultivation. This increase was accompanied by a 3-fold increase of in vitro sucrose-hydrolysis and isomaltase activities, as well as by a 3- to 5-fold upregulation of the isomaltase-encoding genes IMA1 and IMA5. One-step Cas9-mediated deletion of all isomaltase-encoding genes (IMA1-5) completely abolished sucrose hydrolysis. Even after 2 months of incubation, the resulting strain did not grow on sucrose. This sucrose-negative strain can be used as a platform to test metabolic engineering strategies and for fundamental studies into sucrose hydrolysis or transport.
Ämnesord
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
Nyckelord
- disaccharide
- isomaltase
- laboratory evolution
- reverse engineering
- multiple gene deletion
- real-time PCR
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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