Energy coupling of membrane transport and efficiency of sucrose dissimilation in yeast
| العنوان: | Energy coupling of membrane transport and efficiency of sucrose dissimilation in yeast |
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| المؤلفون: | Ryan K Henderson, Sophie C. de Valk, Robert Mans, Bert Poolman |
| المساهمون: | Enzymology |
| المصدر: | Metabolic Engineering, 65, 243-254. ACADEMIC PRESS INC ELSEVIER SCIENCE Metabolic Engineering, 65 |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0106 biological sciences, Sucrose, Saccharomyces cerevisiae Proteins, Saccharomyces cerevisiae, Mutant, Bioengineering, 01 natural sciences, Applied Microbiology and Biotechnology, 03 medical and health sciences, chemistry.chemical_compound, 010608 biotechnology, 030304 developmental biology, 0303 health sciences, Membrane transport, Microbial bioenergetics, biology, Membrane Transport Proteins, Active site, Biological Transport, Directed evolution, Sucrose transport, biology.organism_classification, Energy coupling, Yeast, ATP, chemistry, Biochemistry, Chemostat, Substrate uptake, biology.protein, Biotechnology |
| الوصف: | Proton coupled transport of α-glucosides via Mal11 into Saccharomyces cerevisiae costs one ATP per imported molecule. Targeted mutation of all three acidic residues in the active site resulted in sugar uniport, but expression of these mutant transporters in yeast did not enable growth on sucrose. We then isolated six unique transporter variants of these mutants by directed evolution of yeast for growth on sucrose. In three variants, new acidic residues emerged near the active site that restored proton-coupled sucrose transport, whereas the other evolved transporters still catalysed sucrose uniport. The localization of mutations and transport properties of the mutants enabled us to propose a mechanistic model of proton-coupled sugar transport by Mal11. Cultivation of yeast strains expressing one of the sucrose uniporters in anaerobic, sucrose-limited chemostat cultures indicated an increase in the efficiency of sucrose dissimilation by 21% when additional changes in strain physiology were taken into account. We thus show that a combination of directed and evolutionary engineering results in more energy efficient sucrose transport, as a starting point to engineer yeast strains with increased yields for industrially relevant products. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1096-7176 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7913d67fb8589072f908d6fd82251935 https://research.rug.nl/en/publications/e8a15678-e3c4-43b1-831e-62d41a7e4700 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....7913d67fb8589072f908d6fd82251935 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10967176 |
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