Optimization of Methane Feed and N:C Ratio for Biomass and Polyhydroxybutyrate Production by the Alphaproteobacterial Methanotroph Methylocystis sp. Rockwell
Abstract
:1. Introduction
2. Materials and Methods
2.1. Media Preparation and Bacterial Cultivation
2.2. Composition of Headspace Gases
2.3. Experimental Design and Statistical Analysis
2.4. PHB Quantification
3. Results
3.1. One-Variable-at-a-Time (OVAT) Analysis of Biomass and PHB Production
3.2. Analysis of Response Surface Methodology (RSM) for Biomass and PHB Production
3.3. Validation of Multi-Objective Optimal Conditions (MOOC)
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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KNO3 (mM) | NH4Cl (mM) | CH4 (mmol) | Final OD540 | Dry wt. (mg/L) | PHB (mg/L) | PHB Cell Content (%DW) |
---|---|---|---|---|---|---|
10 | - | 0 | 0.009 ± 0.001 | 5.33 ± 2.31 | 0.00 | 0.00 |
10 | - | 2 | 0.266 ± 0.028 | 157.33 ± 8.33 | 0.00 | 0.00 |
10 | - | 4 | 0.417 ± 0.051 | 270.67 ± 22.03 | 9.16 ± 3.33 | 3.47 ± 1.54 |
10 | - | 6 | 0.596 ± 0.012 | 378.67 ± 43.88 | 41.70 ± 16.51 | 11.20 ± 4.89 |
10 | - | 8 | 0.558 ± 0.098 | 340 ± 24 | 56.75 ± 6.81 | 16.81 ± 2.98 |
10 | - | 10 | 0.550 ± 0.045 | 334.67 ± 32.33 | 35.28 ± 10.21 | 10.45 ± 2.49 |
- | 10 | 0 | 0.014 ± 0.001 | 5.33 ± 2.31 | 0.00 | 0.00 |
- | 10 | 2 | 0.393 ± 0.015 | 232 ± 10.58 | 19.83 ± 1.08 | 8.55 ± 0.16 |
- | 10 | 4 | 0.596 ± 0.003 | 320 ± 20.78 | 50.69 ± 13.72 | 15.90 ± 4.33 |
- | 10 | 6 | 0.679 ± 0.004 | 461.33 ± 26.63 | 12.85 ± 1.31 | 2.86 ± 0.41 |
- | 10 | 8 | 0.626 ± 0.004 | 397.33 ± 8.33 | 12.09 ± 2.34 | 3.04 ± 0.54 |
- | 10 | 10 | 0.601 ± 0.013 | 325.33 ± 33.31 | 11.95 ± 0.92 | 3.72 ± 0.70 |
CH4 (mmol) | NH4Cl (mM) | KNO3 (mM) | Final OD540 | Dry wt. (mg/L) | PHB (mg/L) | PHB Cell Content (%DW) |
---|---|---|---|---|---|---|
6 | - | 0 | 0.153 ± 0.016 | 81.33 ± 14.05 | 24.19 ± 6.60 | 30 ± 7.94 |
6 | - | 0.5 | 0.470 ± 0.013 | 274 ± 8.49 | 96.69 ± 2.67 | 35.32 ± 2.07 |
6 | - | 1 | 0.612 ± 0.025 | 408 ± 6.93 | 65.06 ± 29.82 | 16.03 ± 7.60 |
6 | - | 2 | 0.635 ± 0.015 | 416 ± 33.94 | 18.33 ± 2.82 | 4.39 ± 0.32 |
6 | - | 4 | 0.685 ± 0.045 | 466 ± 59.40 | 7.34 ± 3.24 | 1.63 ± 0.90 |
6 | - | 8 | 0.731 ± 0.012 | 469.33 ± 8.33 | 10.83 ± 1.17 | 2.31 ± 0.29 |
6 | 0 | - | 0.153 ± 0.016 | 81.33 ± 14.05 | 24.19 ± 6.60 | 30 ± 7.94 |
6 | 0.5 | - | 0.424 ± 0.015 | 276 ±10.58 | 113.31 ± 9.78 | 41.15 ± 4.63 |
6 | 1 | - | 0.621 ± 0.050 | 404 ± 31.24 | 196.12 ± 29.75 | 48.54 ± 6.55 |
6 | 2 | - | 0.668 ± 0.015 | 457.33 ± 25.40 | 133.62 ± 26.24 | 29.20 ± 5.24 |
6 | 4 | - | 0.720 ± 0.031 | 462.67 ± 25.72 | 97.15 ± 12.67 | 21 ± 2.37 |
6 | 8 | - | 0.742 ± 0.003 | 465.33 ± 9.24 | 93.33 ± 5.83 | 20.05 ± 1.20 |
Optimization | N-Source | N:C Ratio | Methane (mmol) | Biomass Yield (mg/L) | PHB Yield (mg/L) | PHB Content (% Cell DW) | |||
---|---|---|---|---|---|---|---|---|---|
Projection | Experimental | Projection | Experimental | Projection | Experimental | ||||
1- %PHB cell content, PHB conc. | NMS | 0.016 | 4.88 | 352.46 | 372 ± 20 | 124.44 | 173.65 ± 13.10 | 36.08 | 46.79 ± 4.74 |
AMS | 0.016 | 6.28 | 364.84 | 372 ± 38.15 | 158 | 196.93 ± 12.80 | 44.50 | 53.11 ± 3.03 | |
2- Biomass, %PHB cell content, PHB conc. | NMS | 0.017 | 6.07 | 414.42 | 431.67 ± 34.03 | 130.56 | 102.04 ± 2.88 | 32.58 | 23.73 ± 1.83 |
AMS | 0.019 | 6.88 | 408.66 | 488.33 ± 7.64 | 162.88 | 164.44 ± 3.21 | 41.72 | 33.68 ± 0.79 |
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Sharma, H.K.; Sauvageau, D.; Stein, L.Y. Optimization of Methane Feed and N:C Ratio for Biomass and Polyhydroxybutyrate Production by the Alphaproteobacterial Methanotroph Methylocystis sp. Rockwell. Methane 2022, 1, 355-364. https://doi.org/10.3390/methane1040026
Sharma HK, Sauvageau D, Stein LY. Optimization of Methane Feed and N:C Ratio for Biomass and Polyhydroxybutyrate Production by the Alphaproteobacterial Methanotroph Methylocystis sp. Rockwell. Methane. 2022; 1(4):355-364. https://doi.org/10.3390/methane1040026
Chicago/Turabian StyleSharma, Hem K., Dominic Sauvageau, and Lisa Y. Stein. 2022. "Optimization of Methane Feed and N:C Ratio for Biomass and Polyhydroxybutyrate Production by the Alphaproteobacterial Methanotroph Methylocystis sp. Rockwell" Methane 1, no. 4: 355-364. https://doi.org/10.3390/methane1040026