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Strategies for Efficient Expression of Heterologous Monosaccharide Transporters in Saccharomyces cerevisiae

2022-01Journal article Open access
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dc.contributor.authorKnychala, Marília
dc.contributor.authorSantos, Angela A. dos
dc.contributor.authorKretzer, Leonardo
dc.contributor.authorGelsleichter, Fernanda
dc.contributor.authorLeandro, Maria José
dc.contributor.authorFonseca, César
dc.contributor.authorStambuk, Boris
dc.date.accessioned2022-03-14T15:45:34Z
dc.date.available2022-03-14T15:45:34Z
dc.date.issued2022-01
dc.description.abstractABSTRACT: In previous work, we developed a Saccharomyces cerevisiae strain (DLG-K1) lacking the main monosaccharide transporters (hxt-null) and displaying high xylose reductase, xylitol dehydrogenase and xylulokinase activities. This strain proved to be a useful chassis strain to study new glucose/xylose transporters, as SsXUT1 from Scheffersomyces stipitis. Proteins with high amino acid sequence similarity (78-80%) to SsXUT1 were identified from Spathaspora passalidarum and Spathaspora arborariae genomes. The characterization of these putative transporter genes (SpXUT1 and SaXUT1, respectively) was performed in the same chassis strain. Surprisingly, the cloned genes could not restore the ability to grow in several monosaccharides tested (including glucose and xylose), but after being grown in maltose, the uptake of C-14-glucose and C-14-xylose was detected. While SsXUT1 lacks lysine residues with high ubiquitinylation potential in its N-terminal domain and displays only one in its C-terminal domain, both SpXUT1 and SaXUT1 transporters have several such residues in their C-terminal domains. A truncated version of SpXUT1 gene, deprived of the respective 3 '-end, was cloned in DLG-K1 and allowed growth and fermentation in glucose or xylose. In another approach, two arrestins known to be involved in the ubiquitinylation and endocytosis of sugar transporters (ROD1 and ROG3) were knocked out, but only the rog3 mutant allowed a significant improvement of growth and fermentation in glucose when either of the XUT permeases were expressed. Therefore, for the efficient heterologous expression of monosaccharide (e.g., glucose/xylose) transporters in S. cerevisiae, we propose either the removal of lysines involved in ubiquitinylation and endocytosis or the use of chassis strains hampered in the specific mechanism of membrane protein turnover.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationKnychala, Marília... [et.al.] - Strategies for Efficient Expression of Heterologous Monosaccharide Transporters in Saccharomyces cerevisiae. In: Journal of Fungi, 2022, Vol. 8(1), article nº 84pt_PT
dc.identifier.doi10.3390/jof8010084pt_PT
dc.identifier.eissn2309-608X
dc.identifier.urihttp://hdl.handle.net/10400.9/3807
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherMDPIpt_PT
dc.relationCAPES/FCT (process 359/14)pt_PT
dc.relationCNPq (process 490029/2009-4, 551392/2010-0, 307290/20123, 478841/2013-2, 30862/2015-6 and 308389/2019-0)pt_PT
dc.relationFINEP (process 01.09.0566.00/1421-08)pt_PT
dc.relationEngineering yeast for efficient co-fermentation of mixed sugars: advantages for ethanol production in the food and fuel industries
dc.relation.publisherversionhttps://doi.org/10.3390/jof8010084pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectEthanolpt_PT
dc.subjectXylosept_PT
dc.subjectYeastspt_PT
dc.subjectSugarspt_PT
dc.subjectSaccharomyces cerevisiaept_PT
dc.titleStrategies for Efficient Expression of Heterologous Monosaccharide Transporters in Saccharomyces cerevisiaept_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleEngineering yeast for efficient co-fermentation of mixed sugars: advantages for ethanol production in the food and fuel industries
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F102803%2F2014/PT
oaire.citation.titleJournal of Fungipt_PT
oaire.citation.volume8pt_PT
person.familyNameKnychala
person.familyNameKretzer
person.familyNameTravassos Leandro
person.familyNameFonseca
person.familyNameStambuk
person.givenNameMarília
person.givenNameLeonardo
person.givenNameMaria José
person.givenNameCésar
person.givenNameBoris
person.identifier2018724
person.identifier2013922
person.identifierK-5461-2015
person.identifier284260
person.identifier464730
person.identifier.ciencia-idE91B-D158-3535
person.identifier.orcid0000-0001-6803-5670
person.identifier.orcid0000-0002-6547-1239
person.identifier.orcid0000-0002-2254-4610
person.identifier.orcid0000-0003-2448-7063
person.identifier.orcid0000-0001-7507-1020
person.identifier.ridA-6757-2010
person.identifier.scopus-author-id36854236300
person.identifier.scopus-author-id55955425000
person.identifier.scopus-author-id7004417604
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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