Phenotypic and genomic analysis of Clostridium beijerinckii NRRL B-598 mutants with increased butanol tolerance

Simple item page
dc.contributor.authorVasylkivska, Marynacs
dc.contributor.authorBranská, Barboracs
dc.contributor.authorSedlář, Karelcs
dc.contributor.authorJurečková, Kateřinacs
dc.contributor.authorProvazník, Ivocs
dc.contributor.authorPatáková, Petracs
dc.coverage.issue1cs
dc.coverage.volume8cs
dc.date.accessioned2020-12-08T11:56:21Z
dc.date.available2020-12-08T11:56:21Z
dc.date.issued2020-11-05cs
dc.description.abstractN-Butanol, a valuable solvent and potential fuel extender, can be produced via acetone-butanol-ethanol (ABE) fermentation. One of the main drawbacks of ABE fermentation is the high toxicity of butanol to producing cells, leading to cell membrane disruption, low culture viability and, consequently, low produced concentrations of butanol. The goal of this study was to obtain mutant strains of Clostridium beijerinckii NRRL B-598 with improved butanol tolerance using random chemical mutagenesis, describe changes in their phenotypes compared to the wild-type strain and reveal changes in the genome that explain improved tolerance or other phenotypic changes. Nine mutant strains with stable improved features were obtained by three different approaches and, for two of them, ethidium bromide (EB), a known substrate of efflux pumps, was used for either selection or as a mutagenic agent. It is the first utilization of this approach for the development of butanol-tolerant mutants of solventogenic clostridia, for which generally there is a lack of knowledge about butanol efflux or efflux mechanisms and their regulation. Mutant strains exhibited increase in butanol tolerance from 36 % up to 127 % and the greatest improvement was achieved for the strains for which EB was used as a mutagenic agent. Additionally, increased tolerance to other substrates of efflux pumps, EB and ethanol, was observed in all mutants and higher antibiotic tolerance in some of the strains. The complete genomes of mutant strains were sequenced and revealed that improved butanol tolerance can be attributed to mutations in genes encoding typical stress responses (chemotaxis, autolysis or changes in cell membrane structure), but, also, to mutations in genes X276_07980 and X276_24400, encoding efflux pump regulators. The latter observation confirms the importance of efflux in butanol stress response of the strain and offers new targets for rational strain engineering.en
dc.formattextcs
dc.format.extent1-18cs
dc.format.mimetypeapplication/pdfcs
dc.identifier.citationFrontiers in Bioengineering and Biotechnology. 2020, vol. 8, issue 1, p. 1-18.en
dc.identifier.doi10.3389/fbioe.2020.598392cs
dc.identifier.issn2296-4185cs
dc.identifier.other165803cs
dc.identifier.urihttp://hdl.handle.net/11012/195765
dc.language.isoencs
dc.publisherFrontierscs
dc.relation.ispartofFrontiers in Bioengineering and Biotechnologycs
dc.relation.urihttps://www.frontiersin.org/articles/10.3389/fbioe.2020.598392/fullcs
dc.rightsCreative Commons Attribution 4.0 Internationalcs
dc.rights.accessopenAccesscs
dc.rights.sherpahttp://www.sherpa.ac.uk/romeo/issn/2296-4185/cs
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/cs
dc.subjectbutanol toleranceen
dc.subjectrandom chemical mutagenesisen
dc.subjectsolventogenic Clostridiumen
dc.subjectgenome sequenceen
dc.subjectbutanol effluxen
dc.titlePhenotypic and genomic analysis of Clostridium beijerinckii NRRL B-598 mutants with increased butanol toleranceen
dc.type.driverarticleen
dc.type.statusPeer-revieweden
dc.type.versionpublishedVersionen
sync.item.dbidVAV-165803en
sync.item.dbtypeVAVen
sync.item.insts2020.12.08 12:56:21en
sync.item.modts2020.12.08 12:14:37en
thesis.grantorVysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií. Ústav biomedicínského inženýrstvícs
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
fbioe085983921.pdf
Size:
2.17 MB
Format:
Adobe Portable Document Format
Description:
fbioe085983921.pdf
Download
Collections
Ústav biomedicínského inženýrství