<div class="csl-bib-body">
<div class="csl-entry">Schwaighofer, A., Ablasser, S., Lux, L., Kopp, J., Herwig, C., Spadiut, O., Lendl, B., & Slouka, C. (2020). Production of Active Recombinant Hyaluronidase Inclusion Bodies from Apis mellifera in E. coli Bl21(DE3) and characterization by FT-IR Spectroscopy. <i>International Journal of Molecular Sciences</i>, <i>54</i>(44), 1–14. https://doi.org/10.3390/ijms21113881</div>
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The bacterium E. coli is one of the most important hosts for recombinant protein production. The benefits are high growth rates, inexpensive media, and high protein titers. However, complex proteins with high molecular weight and many disulfide bonds are expressed as inclusion bodies (IBs). In the last decade, the overall perception of these IBs being not functional proteins changed, as enzyme activity was found within IBs. Several applications for direct use of IBs are already reported in literature. While fluorescent proteins or protein tags are used for determination of IB activity to date, direct measurements of IB protein activity are scacre. The expression of recombinant hyaluronidase from Apis mellifera in E. coli BL21(DE3) was analyzed using a face centered design of experiment approach. Hyaluronidase is a hard to express protein and imposes a high metabolic burden to the host. Conditions giving a high specific IB titer were found at 25 °C at low specific substrate uptake rates and induction times of 2 to 4 h. The protein activity of hyaluronidase IBs was verified using (Fourier transform) FT-IR spectroscopy. Degradation of the substrate hyaluronan occurred at increased rates with higher IB concentrations. Active recombinant hyaluronidase IBs can be immediately used for direct degradation of hyaluronan without further down streaming steps. FT-IR spectroscopy was introduced as a method for tracking IB activity and showed differences in degradation behavior of hyaluronan dependent on the applied active IB concentration.
en
dc.description.sponsorship
Fonds zur Förderung der Wissenschaftlichen Forschung
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dc.description.sponsorship
Österreichische Forschungsförderungsgesellschaft
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dc.language
English
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dc.language.iso
en
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dc.publisher
MDPI
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dc.relation.ispartof
International Journal of Molecular Sciences
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dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
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dc.subject
Escherichia coli
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dc.subject
active inclusion body
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dc.subject
recombinant protein production
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dc.subject
upstream process development
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dc.subject
FT-IR spectroscopy
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dc.title
Production of Active Recombinant Hyaluronidase Inclusion Bodies from Apis mellifera in E. coli Bl21(DE3) and characterization by FT-IR Spectroscopy
en
dc.type
Article
en
dc.type
Artikel
de
dc.rights.license
Creative Commons Namensnennung 4.0 International
de
dc.rights.license
Creative Commons Attribution 4.0 International
en
dc.description.startpage
1
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dc.description.endpage
14
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dc.relation.grantno
P32644-N
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dc.relation.grantno
868615
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dc.rights.holder
The Author(s)
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dc.type.category
Original Research Article
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tuw.container.volume
54
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tuw.container.issue
44
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tuw.journal.peerreviewed
true
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tuw.peerreviewed
true
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tuw.version
vor
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dcterms.isPartOf.title
International Journal of Molecular Sciences
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tuw.publication.orgunit
E164 - Institut für Chemische Technologien und Analytik
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tuw.publication.orgunit
E166 - Institut für Verfahrenstechnik, Umwelttechnik und technische Biowissenschaften