Abstract:Data obtained from in situ Raman spectroscopy probes and high‐performance liquid chromatography (HPLC) analysis were applied together with chemometrics to build partial least squares models of metabolite concentrations for the industrially relevant organism Clostridium acetobutylicum. Models were built for predominant products (acetic acid, butyric acid, and butanol) of C. acetobutylicum cultures grown on glucose as a substrate. The partial least squares models were then applied to a 3‐day C. acetobutylicum cu… Show more
“…Clostridium sp. was initially propagated on CGM under anaerobic conditions described previously 28 , 40 . 500 µL of potato-glucose suspension (PGM) spore stock was heat-shocked at 80 °C for 10 min, then re-suspended in 150 mL CGM enriched with 0.5% of the respective carbohydrate (i.e., glucose, galacturonate, gluconate, or mixtures two or three of these substrates).…”
Clostridiumacetobutylicum ATCC 824 effectively utilizes a wide range of substrates to produce commodity chemicals. When grown on substrates of different oxidation states, the organism exhibits different recycling needs of reduced intracellular electron carrying co-factors. Ratios of substrates with different oxidation states were used to modulate the need to balance electron carriers and demonstrate fine-tuned control of metabolic output. Three different oxidized substrates were first fed singularly, then in different ratios to three different strains of Clostridium sp. Growth was most robust when fed glucose in exclusive fermentations. However, the use of the other two more oxidized substrates was strain-dependent in exclusive feeds. In glucose-galacturonate mixed fermentation, the main products (acetate and butyrate) were dependant on the ratios of the substrates. Exclusive fermentation on galacturonate was nearly homoacetic. Co-utilization of galacturonate and glucose was observed from the onset of fermentation in growth conditions using both substrates combined, with the proportion of galacturonate present dictating the amount of acetate produced. For all three strains, increasing galacturonate content (%) in a mixture of galacturonate and glucose from 0 to 50, and 100, resulted in a corresponding increase in the amount of acetate produced. For example, C.acetobutylicum increased from ~ 10 mM to ~ 17 mM, and then ~ 23 mM. No co-utilization was observed when galacturonate was replaced with gluconate in the two substrate co-feed.
“…Clostridium sp. was initially propagated on CGM under anaerobic conditions described previously 28 , 40 . 500 µL of potato-glucose suspension (PGM) spore stock was heat-shocked at 80 °C for 10 min, then re-suspended in 150 mL CGM enriched with 0.5% of the respective carbohydrate (i.e., glucose, galacturonate, gluconate, or mixtures two or three of these substrates).…”
Clostridiumacetobutylicum ATCC 824 effectively utilizes a wide range of substrates to produce commodity chemicals. When grown on substrates of different oxidation states, the organism exhibits different recycling needs of reduced intracellular electron carrying co-factors. Ratios of substrates with different oxidation states were used to modulate the need to balance electron carriers and demonstrate fine-tuned control of metabolic output. Three different oxidized substrates were first fed singularly, then in different ratios to three different strains of Clostridium sp. Growth was most robust when fed glucose in exclusive fermentations. However, the use of the other two more oxidized substrates was strain-dependent in exclusive feeds. In glucose-galacturonate mixed fermentation, the main products (acetate and butyrate) were dependant on the ratios of the substrates. Exclusive fermentation on galacturonate was nearly homoacetic. Co-utilization of galacturonate and glucose was observed from the onset of fermentation in growth conditions using both substrates combined, with the proportion of galacturonate present dictating the amount of acetate produced. For all three strains, increasing galacturonate content (%) in a mixture of galacturonate and glucose from 0 to 50, and 100, resulted in a corresponding increase in the amount of acetate produced. For example, C.acetobutylicum increased from ~ 10 mM to ~ 17 mM, and then ~ 23 mM. No co-utilization was observed when galacturonate was replaced with gluconate in the two substrate co-feed.
“…They used data obtained from in situ Raman spectroscopy probes and HPLC analysis, together with chemometrics, to build partial least square models of metabolite concentrations for the industrially relevant organism C. acetobutylicum . They concluded that predictive models based upon Raman spectral data are promising tools for characterization of synthetic organisms, guiding process control, and facilitating optimization of culture conditions …”
This annual review is an overview of advances in the field of Raman spectroscopy as found in papers published in the previous calendar year in the Journal of Raman Spectroscopy (JRS), as well as in trends over the past decade across journals that have published papers important to the field of Raman spectroscopy. This information is gleaned from statistical data on article counts obtained from the Clarivate Analytic's Web of Science Core Collection by year and by subfield of Raman spectroscopy. Additional information is gleaned from presentations at the XXVI International Conference on Raman Spectroscopy (ICORS 2018) in Jeju Island, Korea in August 2018, and contributions on Raman scattering at SCIX 2018, organized by the Federation of Analytical Chemistry and Spectroscopy Societies (FACSS) in Atlanta, Georgia, USA in October 2018. Coverage is also provided for topics from the European Conference on Nonlinear Optical Spetroscopy (ECONOS 2018) held in April in Milan, Italy and GeoRaman 2018 in Catania, Italy in June. Finally, papers published in JRS in 2017 are highlighted and arranged by topics at the frontier of Raman spectroscopy. On the basis of this survey information, it is clear that Raman spectroscopy continues as in recent years as a rapidly expanding field of research across a wide range of novel disciplines and applications that provide sensitive photonic information of matter at the molecular level.
“…Fermentation monitoring is a pivotal tool for bioprocess optimization and control. Assessing the state of the process via high‐resolution time‐course analysis allows detailed characterization during process development, as well as rapid detection and correction of any possible deviations from desired process specifications during product manufacturing, ensuring the quality of the end product (Svendsen et al, 2015; Zu et al, 2017).…”
Section: Introductionmentioning
confidence: 99%
“…However, these vibrational spectroscopy technologies have certain limitations, the main one being that the spectra that they generate are very convoluted with many overlapping signals. This results in the need to use chemometric mathematical models such as partial least squares (PLS) regression to break down the different signals contributed by the different compounds in the mixture (do Nascimento et al, 2017; Li et al, 2018; Marison et al, 2012; Rodrigues et al, 2018; Stuart, 2005; Zu et al, 2017). These models require significant time and resources to build and are usually not transferable, that is, they are only applicable to the configuration used to build them (bioreactor, medium composition, strain, temperature, pH, etc.)…”
The real-time monitoring of metabolites (RTMet) is instrumental for the industrial production of biobased fermentation products. This study shows the first application of untargeted on-line metabolomics for the monitoring of undiluted fermentation broth samples taken automatically from a 5 L bioreactor every 5 min via flow injection mass spectrometry. The travel time from the bioreactor to the mass spectrometer was 30 s. Using mass spectrometry allows, on the one hand, the direct monitoring of targeted key process compounds of interest and, on the other hand, provides information on hundreds of additional untargeted compounds without requiring previous calibration data. In this study, this technology was applied in an Escherichia coli succinate fermentation process and 886 different m/z signals were monitored, including key process compounds (glucose, succinate, and pyruvate), potential biomarkers of biomass formation such as (R)-2,3-dihydroxy-isovalerate and (R)-2,3-dihydroxy-3-methylpentanoate and compounds from the pentose phosphate pathway and nucleotide metabolism, among others. The main advantage of the RTMet technology is that it allows the monitoring of hundreds of signals without the requirement of developing partial least squares regression models, making it a perfect tool for bioprocess monitoring and for testing many different strains and process conditions for bioprocess development.
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