Comparison of constitutive and inducible β-fructofuranosidase production by recombinant Pichia pastoris in fed-batch culture using defined and semi-defined media

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dc.contributor.authorAnane, Emmanuelen_ZA
dc.contributor.authorVan Rensburg, Eugeneen_ZA
dc.contributor.authorGorgens, Johann F.en_ZA
dc.date.accessioned2017-10-25T12:04:00Z
dc.date.available2017-10-25T12:04:00Z
dc.date.issued2016
dc.descriptionCITATION: Anane, E., Van Rensburg, E. & Gorgens, J. F. 2016. Comparison of constitutive and inducible β-fructofuranosidase production by recombinant Pichia pastoris in fed-batch culture using defined and semi-defined media. South African Journal of Chemical Engineering, 22:17-22, doi:10.1016/j.sajce.2016.10.001.
dc.descriptionThe original publication is available at http://www.sciencedirect.com
dc.description.abstractENGLISH ABSTRACT: Short-chain fructooligosaccharides produced from sucrose by transfructosylation using β-fructofuranosidase (FFase), an industrially important enzyme, finds application in pre-biotics, sweeteners and confectionary products. Using recombinant Pichia pastoris, the influence of replacing the commonly-used Invitrogen® medium with a semi-defined medium for FFase production under the control of the glyceraldehyde-3-phosphate dehydrogenase (GAP) and alcohol oxidase (AOX) promoters was investigated. Replacing the trace metals (PTM1) solution with yeast extract resulted in a 54.3% decrease in FFase volumetric activity under control of the AOX promoter, suggesting a distinct requirement for trace metals for recombinant protein synthesis during methanol induction, given that the biomass yield on methanol decreased by only 10%. The same medium adjustment had no effect on enzyme production under GAP promoter control, although AOX promoter control resulted in double the FFase volumetric activity compared to glycerol-fed cultures. Decreasing basal salts by half did not affect the cultures, but alleviated precipitation during sterilisation. Optimisation of the glycerol feed rate and dissolved oxygen tension in DO-stat fed-batch fermentations using the semi-defined medium resulted in 17% increase in volutmetric activity of FFase expressed under the GAP promoter. This study highlighted the influence of carbon source and trace metals on heterologous protein production by P. pastoris using constitutive and inducible promoters.en_ZA
dc.description.urihttp://www.sciencedirect.com/science/article/pii/S1026918516300026
dc.description.versionPublisher's version
dc.format.extent6 pagesen_ZA
dc.identifier.citationAnane, E., Van Rensburg, E. & Gorgens, J. F. 2016. Comparison of constitutive and inducible β-fructofuranosidase production by recombinant Pichia pastoris in fed-batch culture using defined and semi-defined media. South African Journal of Chemical Engineering, 22:17-22, doi:10.1016/j.sajce.2016.10.001
dc.identifier.issn1026-9185
dc.identifier.otherdoi:10.1016/j.sajce.2016.10.001
dc.identifier.urihttp://hdl.handle.net/10019.1/102384
dc.language.isoen_ZAen_ZA
dc.publisherElsevieren_ZA
dc.rights.holderAuthors retain copyrighten_ZA
dc.subjectPichia pastorisen_ZA
dc.subjectFructofuranosidaseen_ZA
dc.subjectDehydrogenasesen_ZA
dc.titleComparison of constitutive and inducible β-fructofuranosidase production by recombinant Pichia pastoris in fed-batch culture using defined and semi-defined mediaen_ZA
dc.typeArticleen_ZA
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