Raman spectroscopy online monitoring of biomass production, intracellular metabolites and carbon substrates during submerged fermentation of oleaginous and carotenogenic microorganisms

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dc.contributor.authorDzurendová, Simonacs
dc.contributor.authorOlsen, Pernille Margrethecs
dc.contributor.authorByrtusová, Danacs
dc.contributor.authorTafintseva, Valeriacs
dc.contributor.authorShapaval, Volhacs
dc.contributor.authorHorn, Svein Jarlecs
dc.contributor.authorKohler, Achimcs
dc.contributor.authorSzotkowski, Martincs
dc.contributor.authorMárová, Ivanacs
dc.contributor.authorZimmermann, Boriscs
dc.coverage.issue261cs
dc.coverage.volume22cs
dc.date.accessioned2024-02-23T13:45:33Z
dc.date.available2024-02-23T13:45:33Z
dc.date.issued2023-12-18cs
dc.description.abstractBackground: Monitoring and control of both growth media and microbial biomass is extremely important for the development of economical bioprocesses. Unfortunately, process monitoring is still dependent on a limited number of standard parameters (pH, temperature, gasses etc.), while the critical process parameters, such as biomass, product and substrate concentrations, are rarely assessable in-line. Bioprocess optimization and monitoring will greatly benefit from advanced spectroscopy-based sensors that enable real-time monitoring and control. Here, Fourier transform (FT) Raman spectroscopy measurement via flow cell in a recirculatory loop, in combination with predictive data modeling, was assessed as a fast, low-cost, and highly sensitive process analytical technology (PAT) system for online monitoring of critical process parameters. To show the general applicability of the method, submerged fermentation was monitored using two different oleaginous and carotenogenic microorganisms grown on two different carbon substrates: glucose fermentation by yeast Rhodotorula toruloides and glycerol fermentation by marine thraustochytrid Schizochytrium sp. Additionally, the online FT-Raman spectroscopy approach was compared with two at-line spectroscopic methods, namely FT-Raman and FT-infrared spectroscopies in high throughput screening (HTS) setups. Results: The system can provide real-time concentration data on carbon substrate (glucose and glycerol) utilization, and production of biomass, carotenoid pigments, and lipids (triglycerides and free fatty acids). Robust multivariate regression models were developed and showed high level of correlation between the online FT-Raman spectral data and reference measurements, with coefficients of determination (R-2) in the 0.94-0.99 and 0.89-0.99 range for all concentration parameters of Rhodotorula and Schizochytrium fermentation, respectively. The online FT-Raman spectroscopy approach was superior to the at-line methods since the obtained information was more comprehensive, timely and provided more precise concentration profiles. Conclusions: The FT-Raman spectroscopy system with a flow measurement cell in a recirculatory loop, in combination with prediction models, can simultaneously provide real-time concentration data on carbon substrate utilization, and production of biomass, carotenoid pigments, and lipids. This data enables monitoring of dynamic behaviour of oleaginous and carotenogenic microorganisms, and thus can provide critical process parameters for process optimization and control. Overall, this study demonstrated the feasibility of using FT-Raman spectroscopy for online monitoring of fermentation processes.en
dc.formattextcs
dc.format.extent14cs
dc.format.mimetypeapplication/pdfcs
dc.identifier.citationMicrobial Cell Factories. 2023, vol. 22, issue 261, 14 p.en
dc.identifier.doi10.1186/s12934-023-02268-ycs
dc.identifier.issn1475-2859cs
dc.identifier.orcid0000-0001-8286-2349cs
dc.identifier.orcid0000-0002-2095-1837cs
dc.identifier.other187228cs
dc.identifier.researcheridA-6897-2016cs
dc.identifier.scopus6603465110cs
dc.identifier.urihttps://hdl.handle.net/11012/245209
dc.language.isoencs
dc.publisherBMCcs
dc.relation.ispartofMicrobial Cell Factoriescs
dc.relation.urihttps://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-023-02268-ycs
dc.rightsCreative Commons Attribution 4.0 Internationalcs
dc.rights.accessopenAccesscs
dc.rights.sherpahttp://www.sherpa.ac.uk/romeo/issn/1475-2859/cs
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/cs
dc.subjectProcess analytical technologyen
dc.subjectRaman spectroscopyen
dc.subjectInfrared spectroscopyen
dc.subjectReal-time monitoringen
dc.subjectPartial least squares (PLS) regressionen
dc.subjectLipidsen
dc.subjectCarotenoidsen
dc.subjectRhodotorulaen
dc.subjectSchizochytriumen
dc.titleRaman spectroscopy online monitoring of biomass production, intracellular metabolites and carbon substrates during submerged fermentation of oleaginous and carotenogenic microorganismsen
dc.type.driverarticleen
dc.type.statusPeer-revieweden
dc.type.versionpublishedVersionen
sync.item.dbidVAV-187228en
sync.item.dbtypeVAVen
sync.item.insts2024.02.23 14:45:33en
sync.item.modts2024.02.23 14:13:17en
thesis.grantorVysoké učení technické v Brně. Fakulta chemická. Ústav chemie potravin a biotechnologiícs
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