In situ multi-wavelength fluorescence spectroscopy as effective tool to simultaneously monitor spore germination, metabolic activity and quantitative protein production in recombinant Aspergillus niger fed-batch cultures.

2.50
Hdl Handle:
http://hdl.handle.net/10033/19673
Title:
In situ multi-wavelength fluorescence spectroscopy as effective tool to simultaneously monitor spore germination, metabolic activity and quantitative protein production in recombinant Aspergillus niger fed-batch cultures.
Authors:
Ganzlin, Markus; Marose, Stefan; Lu, Xin; Hitzmann, Bernd; Scheper, Thomas; Rinas, Ursula
Abstract:
The production of a mutant green fluorescent protein (S65TGFP), controlled by the maltose inducible glucoamylase promoter, was followed in situ in fed-batch cultures of recombinant Aspergillus niger using multi-wavelength fluorescence spectroscopy. Disturbance of quantitative product analysis by interfering fluorescence signals was resolved by using a set of defined combinations of excitation and emission wavelengths (lambda(ex)/lambda(em)). This technique resulted in excellent linearity between on-line signal and off-line determined S65TGFP concentrations. Spore germination was detectable in situ by monitoring the back scattered light intensity. Moreover, flavin-like fluorophores were identified as the dominating fungal host fluorophores. The time-dependent intensity of this fluorophore, potentially fungal flavin-containing oxidoreductase(s), did not correlate with the biomass concentration but correlated well with the fungal metabolic activity (e.g. respiratory activity). Other fluorophores commonly found in microbial cultures such NADH, pyridoxine and the aromatic amino acids, tryptophan, phenylalanine and tyrosine did not contribute significantly to the culture fluorescence of A. niger. Thus, multi-wavelength fluorescence spectroscopy has proven to be an effective tool for simultaneous on-line monitoring of the most relevant process variables in fungal cultures, e.g. spore germination, metabolic activity, and quantitative product formation.
Affiliation:
Helmholtz Centre for Infection Research (former German Research Centre for Biotechnology - GBF), Inhoffenstr. 7, 38124 Braunschweig, Germany.
Citation:
In situ multi-wavelength fluorescence spectroscopy as effective tool to simultaneously monitor spore germination, metabolic activity and quantitative protein production in recombinant Aspergillus niger fed-batch cultures. 2007, 132 (4):461-8 J. Biotechnol.
Journal:
Journal of biotechnology
Issue Date:
1-Dec-2007
URI:
http://hdl.handle.net/10033/19673
DOI:
10.1016/j.jbiotec.2007.08.032
PubMed ID:
17905460
Type:
Article
Language:
en
ISSN:
0168-1656
Appears in Collections:
Publications from Division of Molekulare Struktur Biologie (MOSB)

Full metadata record

DC FieldValue Language
dc.contributor.authorGanzlin, Markus-
dc.contributor.authorMarose, Stefan-
dc.contributor.authorLu, Xin-
dc.contributor.authorHitzmann, Bernd-
dc.contributor.authorScheper, Thomas-
dc.contributor.authorRinas, Ursula-
dc.date.accessioned2008-03-04T14:26:26Z-
dc.date.available2008-03-04T14:26:26Z-
dc.date.issued2007-12-01-
dc.identifier.citationIn situ multi-wavelength fluorescence spectroscopy as effective tool to simultaneously monitor spore germination, metabolic activity and quantitative protein production in recombinant Aspergillus niger fed-batch cultures. 2007, 132 (4):461-8 J. Biotechnol.en
dc.identifier.issn0168-1656-
dc.identifier.pmid17905460-
dc.identifier.doi10.1016/j.jbiotec.2007.08.032-
dc.identifier.urihttp://hdl.handle.net/10033/19673-
dc.description.abstractThe production of a mutant green fluorescent protein (S65TGFP), controlled by the maltose inducible glucoamylase promoter, was followed in situ in fed-batch cultures of recombinant Aspergillus niger using multi-wavelength fluorescence spectroscopy. Disturbance of quantitative product analysis by interfering fluorescence signals was resolved by using a set of defined combinations of excitation and emission wavelengths (lambda(ex)/lambda(em)). This technique resulted in excellent linearity between on-line signal and off-line determined S65TGFP concentrations. Spore germination was detectable in situ by monitoring the back scattered light intensity. Moreover, flavin-like fluorophores were identified as the dominating fungal host fluorophores. The time-dependent intensity of this fluorophore, potentially fungal flavin-containing oxidoreductase(s), did not correlate with the biomass concentration but correlated well with the fungal metabolic activity (e.g. respiratory activity). Other fluorophores commonly found in microbial cultures such NADH, pyridoxine and the aromatic amino acids, tryptophan, phenylalanine and tyrosine did not contribute significantly to the culture fluorescence of A. niger. Thus, multi-wavelength fluorescence spectroscopy has proven to be an effective tool for simultaneous on-line monitoring of the most relevant process variables in fungal cultures, e.g. spore germination, metabolic activity, and quantitative product formation.en
dc.language.isoenen
dc.subject.meshAspergillus nigeren
dc.subject.meshBioreactorsen
dc.subject.meshGerminationen
dc.subject.meshGreen Fluorescent Proteinsen
dc.subject.meshIndustrial Microbiologyen
dc.subject.meshSpectrometry, Fluorescenceen
dc.subject.meshSpores, Fungalen
dc.titleIn situ multi-wavelength fluorescence spectroscopy as effective tool to simultaneously monitor spore germination, metabolic activity and quantitative protein production in recombinant Aspergillus niger fed-batch cultures.en
dc.typeArticleen
dc.contributor.departmentHelmholtz Centre for Infection Research (former German Research Centre for Biotechnology - GBF), Inhoffenstr. 7, 38124 Braunschweig, Germany.en
dc.identifier.journalJournal of biotechnologyen
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