Microbial Conversion of Cheese Whey to Polyhydroxybutyrate (PHB) via Statistically Optimized Cultures
Abstract
:1. Introduction
2. Materials and Methods
2.1. Fermentative PHB Production from Cheese Whey
2.2. Design of Experiments (DoE)
2.3. Analytical Methods
3. Results and Discussion
3.1. Evaluation of Cheese Whey Conversion to PHB
Sugar/ Substrate | OD 1 (@ 600 nm) | DCW 2 (g/L) | CPHB 3 (g/L) | %PHB 4 (% w/w) | CN 5 (g/L) | CS 6 (g/L) | Mw 7 (kDa) |
---|---|---|---|---|---|---|---|
Saccharose (S) | 22.3 ± 2.7 | 6.52 ± 0.7 | 4.11 ± 0.4 | 63.1 ± 5.3 | 0.12 ± 0.04 | 2.57 ± 0.6 | 852 ± 84 |
Glucose (G) | 25 ± 2.6 | 6.25 ± 0.7 | 4.06 ± 0.5 | 65 ± 4.9 | 0.04 ± 0.02 | 1.76 ± 0.4 | 803 ± 81 |
Lactose (L) | 15.4 ± 1.9 | 2.95 ± 0.6 | 1.09 ± 0.3 | 36.9 ± 4.4 | 0.37 ± 0.02 | 4.32 ± 0.7 | 826 ± 66 |
Xylose (X) | 16.4 ± 1.7 | 3.15 ± 0.5 | 1.32 ± 0.4 | 41.9 ± 3.9 | 0.26 ± 0.01 | 5.34 ± 0.5 | 867 ± 74 |
Fructose (F) | 19.1 ± 1.5 | 4.75 ± 0.6 | 2.42 ± 0.6 | 50.9 ± 5.1 | 0.13 ± 0.02 | 2.73 ± 0.5 | 795 ± 53 |
Galactose (GL) | 20.3 ± 2.1 | 5.43 ± 0.8 | 2.41 ± 0.7 | 54.4 ± 4.7 | 0.11 ± 0.01 | 2.48 ± 0.6 | 812 ± 65 |
Glucose + Galactose (GG) | 22.4 ± 2.0 | 6.11 ± 0.7 | 3.46 ± 0.6 | 56.6 ± 3.6 | 0.09 ± 0.03 | 1.54 ± 0.4 | 811 ± 58 |
Cheese Whey (CW) | 18.3 ± 2.2 | 3.24 ± 0.5 | 1.33 ± 0.5 | 41 ± 4.7 | 0.44 ± 0.08 | 7.43 ± 0.3 | – |
Enzymatically Hydrolyzed CW (CWE) | 21.6 ± 2.3 | 6.02 ± 0.6 | 3.33 ± 0.5 | 55.3 ± 6.2 | 0.14 ± 0.06 | 1.78 ± 0.6 | 801 ± 59 |
Acid-Hydrolyzed CW (CWA) | 20.1 ± 2.4 | 5.88 ± 0.6 | 3.25 ± 0.3 | 55.3 ± 4.1 | 0.17 ± 0.03 | 1.89 ± 0.3 | 789 ± 66 |
3.2. DoE, Optimization, and Analysis of Variance
Run # | C/N 1 (w/w) | C/P 2 (w/w) | DO 3 (%) | RT 4 (h) | %PHB 5 (% w/w) | Mw 6 (kDa) |
---|---|---|---|---|---|---|
1 | 6.6 | 0.8 | 5 | 0 | 48.2 ± 4.3 | 668 ± 69 |
2 | 6.6 | 1.9 | 10 | 1 | 54.3 ± 5.3 | 742 ± 79 |
3 | 6.6 | 3 | 20 | 3 | 53.3 ± 5.4 | 717 ± 81 |
4 | 6.6 | 5 | 30 | 9 | 46.7 ± 4.9 | 701 ± 82 |
5 | 8 | 0.8 | 10 | 3 | 52.4 ± 4.7 | 702 ± 74 |
6 | 8 | 1.9 | 5 | 9 | 50.7 ± 5.2 | 669 ± 73 |
7 | 8 | 3 | 30 | 0 | 55.6 ± 6.1 | 756 ± 82 |
8 | 8 | 5 | 20 | 1 | 56.4 ± 5.9 | 782 ± 76 |
9 | 10 | 0.8 | 20 | 9 | 55.8 ± 4.9 | 704 ± 68 |
10 | 10 | 1.9 | 30 | 3 | 56.9 ± 5.1 | 732 ± 73 |
11 | 10 | 3 | 5 | 1 | 55.2 ± 5.2 | 821 ± 82 |
12 | 10 | 5 | 10 | 0 | 54.2 ± 5.7 | 851 ± 85 |
13 | 13.3 | 0.8 | 30 | 1 | 51.2 ± 5.5 | 830 ± 78 |
14 | 13.3 | 1.9 | 20 | 0 | 56.6 ± 5.1 | 892 ± 85 |
15 | 13.3 | 3 | 10 | 9 | 50.2 ± 4.9 | 789 ± 68 |
16 | 13.3 | 5 | 5 | 3 | 46.6 ± 4.7 | 802 ± 73 |
- Carbon-to-nitrogen mass ratio (C/N): 10 w/w for %PHB and 13.3 w/w for Mw.
- Carbon-to-phosphorous mass ratio (C/P): 1.9 w/w for %PHB and 5 w/w for Mw.
- Dissolved oxygen concentration (DO): 20% for both %PHB and Mw.
- Residence time in stationary phase (RT): 1 h for both %PHB and Mw.
- For C/N = 10 w/w, C/P = 1.9 w/w, DO = 20%, and RT = 1 h, the maximum estimated biopolymer content value is %PHB = 61.66% w/w; this value was verified with minimal deviation (2%) by an additional experiment.
- For C/N = 13.3 w/w, C/P = 5 w/w, DO = 20%, and RT = 1 h, the maximum estimated weight average molecular weight value is Mw = 900 kDa; this value was also verified with a small deviation (5%) by a second additional experiment.
3.3. Regression Modeling and Sensitivity Analysis
4. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Scenario | Parameter | C/N 1 | C/P 2 | DO 3 | RT 4 |
---|---|---|---|---|---|
Maximize %PHB 5 | Optimal Level | 10 w/w | 1.9 w/w | 20% | 1 h |
%PHB Range of Variation (Delta, % w/w) | 4.9% | 3.65% | 5.35% | 3.42% | |
Significance Ranking | 2 | 3 | 1 | 4 | |
Contribution to Variance (%) | 28% | 21% | 31% | 20% | |
Maximize Mw 6 | Optimal Level | 13.3 w/w | 5 w/w | 20% | 1 h |
Mw Range of Variation (Delta, kDa) | 121.3 | 58 | 33.8 | 78 | |
Significance Ranking | 1 | 3 | 4 | 2 | |
Contribution to Variance (%) | 42% | 20% | 12% | 26% |
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Penloglou, G.; Pavlou, A.; Kiparissides, C. Microbial Conversion of Cheese Whey to Polyhydroxybutyrate (PHB) via Statistically Optimized Cultures. Fermentation 2023, 9, 624. https://doi.org/10.3390/fermentation9070624
Penloglou G, Pavlou A, Kiparissides C. Microbial Conversion of Cheese Whey to Polyhydroxybutyrate (PHB) via Statistically Optimized Cultures. Fermentation. 2023; 9(7):624. https://doi.org/10.3390/fermentation9070624
Chicago/Turabian StylePenloglou, Giannis, Alexandros Pavlou, and Costas Kiparissides. 2023. "Microbial Conversion of Cheese Whey to Polyhydroxybutyrate (PHB) via Statistically Optimized Cultures" Fermentation 9, no. 7: 624. https://doi.org/10.3390/fermentation9070624