Quantitative Raman Spectroscopy Analysis of Polyhydroxyalkanoates Produced by Cupriavidus necator H16
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microorganisms and Their Cultivation
2.2. Bacterial Sample Preparation and Estimated Time Frame for Analysis
- (a)
- One milliliter of bacterial culture was transferred into a 1.5 mL tube, and the cells were centrifuged (10,000 rpm, 2 min), washed with 50% ethanol, and centrifuged again (total 5 min).
- (b)
- In the next step, the supernatant was aspirated, and approx. 20 µL of cell pellets formed via centrifugation was pipetted onto a CaF (Raman grade) microscope slide (1 min).
- (c)
- The cell suspensions were air-dried (approx. 5 min) at laboratory temperature and analyzed using a Raman instrument (1 min).
2.3 Raman Spectroscopy
2.4. Data Analysisis
3. Results and Discussion
4. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Cultivation Conditions | PHB % | Intensity of PHB Line at 1736 cm−1 (Normalized on Amide I at 1662 cm–1) | Intensity of PHB Line at 1736 cm–1 (Normalized on DNA at 785 cm–1) |
---|---|---|---|
Cupriavidus necator H16 | 12.1 | 0.17 | 0.39 |
MS medium, 5 g/L fructose, 1 g/L (NH4)2SO4, from Petri dish, 72 h of cultivation | |||
Cupriavidus necator H16 | 26.1 | 0.55 | 2.0 |
MS medium, 5 g/L fructose, 3 g/L (NH4)2SO4, 72 h of cultivation | |||
Cupriavidus necator H16 | 30.6 | 0.65 | 2.7 |
NB medium, 24 h of cultivation | |||
Cupriavidus necator H16 | 59.1 | 1.8 | 5.8 |
MS medium, 20 g/L fructose, 1 g/L (NH4)2SO4, 24 h of cultivation | |||
Cupriavidus necator H16 | 65.4 | 2.0 | 7.0 |
MS medium, 20 g/L fructose, 3 g/L (NH4)2SO4, 24 h of cultivation | |||
Cupriavidus necator H16 | 67.7 | 2.2 | 7.2 |
MS medium, 20 g/L fructose, 3 g/l (NH4)2SO4, 24 h of cultivation | |||
Cupriavidus necator H16 | 69.2 | 2.2 | 7.3 |
MS medium, 20 g/L fructose, 3 g/L (NH4)2SO4, 72 h of cultivation | |||
Cupriavidus necator H16 | 90.0 | 3.2 | 9.4 |
MS medium, 20 g/L fructose, 3 g/L (NH4)2SO4, 72 h of cultivation |
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Samek, O.; Obruča, S.; Šiler, M.; Sedláček, P.; Benešová, P.; Kučera, D.; Márova, I.; Ježek, J.; Bernatová, S.; Zemánek, P. Quantitative Raman Spectroscopy Analysis of Polyhydroxyalkanoates Produced by Cupriavidus necator H16. Sensors 2016, 16, 1808. https://doi.org/10.3390/s16111808
Samek O, Obruča S, Šiler M, Sedláček P, Benešová P, Kučera D, Márova I, Ježek J, Bernatová S, Zemánek P. Quantitative Raman Spectroscopy Analysis of Polyhydroxyalkanoates Produced by Cupriavidus necator H16. Sensors. 2016; 16(11):1808. https://doi.org/10.3390/s16111808
Chicago/Turabian StyleSamek, Ota, Stanislav Obruča, Martin Šiler, Petr Sedláček, Pavla Benešová, Dan Kučera, Ivana Márova, Jan Ježek, Silva Bernatová, and Pavel Zemánek. 2016. "Quantitative Raman Spectroscopy Analysis of Polyhydroxyalkanoates Produced by Cupriavidus necator H16" Sensors 16, no. 11: 1808. https://doi.org/10.3390/s16111808
APA StyleSamek, O., Obruča, S., Šiler, M., Sedláček, P., Benešová, P., Kučera, D., Márova, I., Ježek, J., Bernatová, S., & Zemánek, P. (2016). Quantitative Raman Spectroscopy Analysis of Polyhydroxyalkanoates Produced by Cupriavidus necator H16. Sensors, 16(11), 1808. https://doi.org/10.3390/s16111808