Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-4803
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dc.contributor.authorLooser, Verena-
dc.contributor.authorLüthy, Dominik-
dc.contributor.authorStraumann, Marcel-
dc.contributor.authorHecht, Katrin-
dc.contributor.authorMelzoch, Karel-
dc.contributor.authorKovar, Karin-
dc.date.accessioned2018-09-20T09:50:23Z-
dc.date.available2018-09-20T09:50:23Z-
dc.date.issued2017-01-27-
dc.identifier.issn0175-7598de_CH
dc.identifier.issn1432-0614de_CH
dc.identifier.urihttps://digitalcollection.zhaw.ch/handle/11475/10842-
dc.description.abstractAs Pichia pastoris (syn. Komagataella sp.) yeast can secrete pure recombinant proteins at high rates, it is a desirable production system. The function of a novel synthetic variant of the AOX1 promoter was characterised comprehensively using a strain secreting Candida antarctica lipase B (CALB) as a model. A new time-saving approach was introduced to determine, in only one experiment, the hitherto unknown relationship between specific product formation rate (qp) and specific growth rate (μ). Tight control of recombinant protein formation was possible in the absence of methanol, while using glycerol as a sole carbon/energy source. CALB was not synthesised during batch cultivation in excess glycerol (>10 g l-1) and at a growth rate close to μmax (0.15 h−1). Between 0.017 and 0.115 h−1 in glycerol-limited fedbatch cultures, basal levels of qp > 0.4 mg g−1 h−1 CALB were reached, independent of the μ at which the culture grew. At μ > 0.04 h−1, an elevated qp occurred temporarily during the first 20 h after changing to fedbatch mode and decreased thereafter to basal. In order to accelerate the determination of the qp(μ) relationship (kinetics of product formation), the entire μ range was covered in a single fedbatch experiment. By linearly increasing and decreasing glycerol addition rates, μ values were repeatedly shifted from 0.004 to 0.074 h−1 and vice versa. Changes in qp were related to changes in μ. A rough estimation of μ range suitable for production was possible in a single fedbatch, thus significantly reducing the experimental input over previous approaches comprising several experiments.de_CH
dc.language.isoende_CH
dc.publisherSpringerde_CH
dc.relation.ispartofApplied Microbiology and Biotechnologyde_CH
dc.rightshttps://creativecommons.org/licenses/by/4.0/de_CH
dc.subjectSecretionde_CH
dc.subjectPichia pastorisde_CH
dc.subjectMethanol-freede_CH
dc.subjectCALBde_CH
dc.subject.ddc570: Biologiede_CH
dc.titleEffects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris : functional characterisation of a novel promoterde_CH
dc.typeBeitrag in wissenschaftlicher Zeitschriftde_CH
dcterms.typeTextde_CH
zhaw.departementLife Sciences und Facility Managementde_CH
zhaw.organisationalunitInstitut für Chemie und Biotechnologie (ICBT)de_CH
dc.identifier.doi10.21256/zhaw-4803-
dc.identifier.doi10.1007/s00253-017-8123-xde_CH
zhaw.funding.euNode_CH
zhaw.issue8de_CH
zhaw.originated.zhawYesde_CH
zhaw.pages.end3176de_CH
zhaw.pages.start3163de_CH
zhaw.publication.statuspublishedVersionde_CH
zhaw.volume101de_CH
zhaw.publication.reviewPeer review (Publikation)de_CH
zhaw.webfeedBiokatalysede_CH
Appears in collections:Publikationen Life Sciences und Facility Management

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Looser, V., Lüthy, D., Straumann, M., Hecht, K., Melzoch, K., & Kovar, K. (2017). Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris : functional characterisation of a novel promoter. Applied Microbiology and Biotechnology, 101(8), 3163–3176. https://doi.org/10.21256/zhaw-4803
Looser, V. et al. (2017) ‘Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris : functional characterisation of a novel promoter’, Applied Microbiology and Biotechnology, 101(8), pp. 3163–3176. Available at: https://doi.org/10.21256/zhaw-4803.
V. Looser, D. Lüthy, M. Straumann, K. Hecht, K. Melzoch, and K. Kovar, “Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris : functional characterisation of a novel promoter,” Applied Microbiology and Biotechnology, vol. 101, no. 8, pp. 3163–3176, Jan. 2017, doi: 10.21256/zhaw-4803.
LOOSER, Verena, Dominik LÜTHY, Marcel STRAUMANN, Katrin HECHT, Karel MELZOCH und Karin KOVAR, 2017. Effects of glycerol supply and specific growth rate on methanol-free production of CALB by P. pastoris : functional characterisation of a novel promoter. Applied Microbiology and Biotechnology. 27 Januar 2017. Bd. 101, Nr. 8, S. 3163–3176. DOI 10.21256/zhaw-4803
Looser, Verena, Dominik Lüthy, Marcel Straumann, Katrin Hecht, Karel Melzoch, and Karin Kovar. 2017. “Effects of Glycerol Supply and Specific Growth Rate on Methanol-Free Production of CALB by P. Pastoris : Functional Characterisation of a Novel Promoter.” Applied Microbiology and Biotechnology 101 (8): 3163–76. https://doi.org/10.21256/zhaw-4803.
Looser, Verena, et al. “Effects of Glycerol Supply and Specific Growth Rate on Methanol-Free Production of CALB by P. Pastoris : Functional Characterisation of a Novel Promoter.” Applied Microbiology and Biotechnology, vol. 101, no. 8, Jan. 2017, pp. 3163–76, https://doi.org/10.21256/zhaw-4803.


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