Simultaneous Lipid and Carotenoid Production via Rhodotorula paludigena CM33 Using Crude Glycerol as the Main Substrate: Pilot-Scale Experiments
Abstract
:1. Introduction
2. Results and Discussion
2.1. Characterization of Crude Glycerol
2.2. Media Optimization for Carotenoid and Lipid Production in R. paludigena CM33
2.3. Effects of Different Fermentations on Lipid and Carotenoid Productions Using a 500 L Fermenter
2.4. Carotenoids Purification with High-Pressure Homogenization (HPH) and Preparative HPLC and the Analysis by HPLC, LC-MS, and NMR
3. Materials and Methods
3.1. Microorganism and Culture Conditions
3.2. Characteristics of Chemical Composition and Elemental Analysis of Crude Glycerol
3.3. Response Surface Methodology (RSM)—Box–Behnken Designs
3.4. Cultivation of R. paludigena CM33 Using a 500 L Fermenter
3.5. Carotenoids Purification with High-Pressure Homogenizer and Preparative High-Performance Liquid Chromatography
3.6. Analytical Methods
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Parameter | Crude Glycerol | Unit |
---|---|---|
Free glycerol | 58 | % |
pH | 2.38 | - |
C | 25.909 | % |
H | 7.440 | % |
N | <1 | % |
S | 4.139 | % |
Na | 11,769 ± 0.10 | ppm |
Mg | 31.29 ± 0.12 | ppm |
Al | ND | ppm |
P | 38.7 ± 0.50 | ppm |
K | 118.8 ± 0.13 | ppm |
Ca | 2050.080 ± 0.14 | ppm |
Mn | 3.22 ± 0.42 | ppm |
Fe | 31.6 ± 0.34 | ppm |
Co | ND | ppm |
Cu | 4.960 ± 0.14 | ppm |
Zn | 25.79 ± 0.31 | ppm |
Source | Sum of Squares | Degree of Freedom | Mean Square | F-Value | p-Value | |
---|---|---|---|---|---|---|
Carotenoid concentration (ug/L) | ||||||
Model | 257,800 | 9 | 28,639.59 | 216.78 | <0.0001 | significant |
A | 79,600.50 | 1 | 79,600.50 | 602.51 | <0.0001 | significant |
B | 798.47 | 1 | 798.47 | 6.04 | 0.0573 | not significant |
C | 21.90 | 1 | 21.90 | 0.1657 | 0.7008 | not significant |
Residual | 660.58 | 5 | 132.12 | – | – | |
Lack of Fit | 454.58 | 3 | 151.53 | 1.47 | 0.4291 | not significant |
Pure Error | 206.00 | 2 | – | – | ||
Cor Total | 258400 | 14 | – | – | – | |
R2 = 0.9974; R2adj = 0.9928 | ||||||
Lipid content (%) | ||||||
Model | 4570.69 | 9 | 507.85 | 104.44 | <0.0001 | significant |
A | 1891.13 | 1 | 1891.13 | 388.89 | <0.0001 | significant |
B | 50.79 | 1 | 50.79 | 10.44 | 0.0232 | significant |
C | 1.11 | 1 | 1.11 | 0.2277 | 0.6534 | not significant |
Residual | 24.31 | 5 | 4.86 | – | – | |
Lack of Fit | 20.54 | 3 | 6.85 | 3.63 | 0.2234 | not significant |
Pure Error | 3.77 | 2 | – | – | ||
Cor Total | 4595.00 | 14 | – | – | – | |
R2 = 0.9947; R2adj = 0.9852 |
Processes | Volume | OD (600 nm) | Biomass (g/L) | Lipid Content (%) | Carotenoids (mg/g CDW) | Carotenoids (mg/L) |
---|---|---|---|---|---|---|
Batch | 350 L | 15.94 | 9.27 | 40.99 | 1.27 | 11.80 |
Repeated-batch | 350 L | |||||
Batch 1 | 19.35 | 9.40 | 40.95 | 1.89 | 17.77 | |
Batch 2 | 27.92 | 16.81 | 36.04 | 5.69 | 95.65 | |
Batch 3 | 63.72 | 32.33 | 32.55 | 12.99 | 419.97 | |
Batch 4 | 71.76 | 38.25 | 38.22 | 14.64 | 559.98 | |
Total 1330 L | = 24.20 | = 36.94 | = 8.80 | = 273.34 | ||
Fed-batch | 350 L | |||||
1st pulse | 16.54 | 9.98 | 12.08 | 3.36 | 33.53 | |
2nd pulse | 25.54 | 15.38 | 44.48 | 5.20 | 79.98 | |
3rd pulse | 62.12 | 37.37 | 26.73 | 12.67 | 473.48 | |
4th pulse | 75.45 | 45.38 | 38.78 | 15.39 | 698.40 |
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Sriphuttha, C.; Boontawan, P.; Boonyanan, P.; Ketudat-Cairns, M.; Boontawan, A. Simultaneous Lipid and Carotenoid Production via Rhodotorula paludigena CM33 Using Crude Glycerol as the Main Substrate: Pilot-Scale Experiments. Int. J. Mol. Sci. 2023, 24, 17192. https://doi.org/10.3390/ijms242417192
Sriphuttha C, Boontawan P, Boonyanan P, Ketudat-Cairns M, Boontawan A. Simultaneous Lipid and Carotenoid Production via Rhodotorula paludigena CM33 Using Crude Glycerol as the Main Substrate: Pilot-Scale Experiments. International Journal of Molecular Sciences. 2023; 24(24):17192. https://doi.org/10.3390/ijms242417192
Chicago/Turabian StyleSriphuttha, Cheeranan, Pailin Boontawan, Pasama Boonyanan, Mariena Ketudat-Cairns, and Apichat Boontawan. 2023. "Simultaneous Lipid and Carotenoid Production via Rhodotorula paludigena CM33 Using Crude Glycerol as the Main Substrate: Pilot-Scale Experiments" International Journal of Molecular Sciences 24, no. 24: 17192. https://doi.org/10.3390/ijms242417192