EMS-Induced Mutagenesis of Clostridium carboxidivorans for Increased Atmospheric CO2 Reduction Efficiency and Solvent Production
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strain, Media, and Culture Conditions
2.2. Ethyl Methanesulfonate (EMS) Mutagenesis of the P7 Population
2.3. End Product Analysis
2.4. CO2 Reduction and Statistical Analysis
3. Results
3.1. Cell Growth and Product Formation of the Wild Type P7 Strain
3.2. Development of an EMS Mutagenized Population of C. carboxidivorans P7
3.3. P7-EMS Mutagenized Mutants Showed Significant Increased Levels of Alcohol Production
3.4. P7-EMS Mutants Showed Increased Levels of Atmospheric CO2 Reduction
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Availability of data and materials
Ethics and consent to participate
References
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Component | Formula | 1754-A (Original PETC Medium) | 1754-B (Modified PETC Medium) |
---|---|---|---|
Volume (0.5 L) | Volume (0.5 L) | ||
Resazurin | |||
Resazurin | C12H6NO4Na | 200 µL | 200 µL |
Minerals | |||
Ammonium chloride | NH4Cl | 500 mg | 500 mg |
Magnesium sulfate | MgSO4·7H2O | 100 mg | 100 mg |
Potassium chloride | KCl | 50 mg | 50 mg |
Potassium phosphate | KH2PO4 | 50 mg | 50 mg |
Sodium chloride | NaCl | 400 mg | 400 mg |
Calcium chloride | CaCl2·2H2O | 20 mg | Not added |
Buffers | |||
HEPES | C8H18N2O4S | Undefined | 1.8 g |
Carbon source | |||
Yeast Extract | 500 mg | Not added | |
Fructose | C6H12O6 | 2.5 g | Not added |
Sodium Bicarbonate | NaHCO3 | 1 g | 1 g |
Minerals and Vitamins | |||
ATCC Minerals | 10 mL | 1 mL | |
ATCC Vitamins | 10 mL | 1 mL | |
Cysteine/Sulfide | |||
L-Cysteine | C3H7NO2S | 2 g | 2 g |
Sodium sulfide | Na2S·9H2O | 2 g | 2 g |
End Product | P7 Strains | Average (g/L) | End Product Content Increase | Maximum Production (g/L) | End Product Converted on (g/L Abs) | CO2 Reduction (g/Abs) | CO2 Reduction Increase |
---|---|---|---|---|---|---|---|
Ethanol | III-J | 0.77 ± 0.20 * | 3.50X | 0.87 | 4.58 | 0.218 | 8.72 X |
III-R | 1.03 ± 0.40 * | 4.68X | 1.64 | 4.21 | 0.200 | 8 X | |
III-P | 0.91 ± 0.58 * | 4.13X | 1.79 | 3.38 | 0.161 | 6.44 X | |
P7-WT | 0.21 ± 0.01 | _ | 0.22 | 0.40 | 0.025 | _ | |
Butyrate | III-J | 0.54 ± 0.10** | 3.85X | 0.7 | 3.33 | 0.166 | 8.73 X |
III-R | 0.45 ± 0.07** | 3.21X | 0.49 | 2.88 | 0.143 | 7.52 X | |
III-P | 0.27 ± 0.01 * | 1.92X | 0.28 | 1.12 | 0.055 | 2.89 X | |
P7-WT | 0.13 ± 0.01 | _ | 0.14 | 0.50 | 0.019 | _ | |
Butanol | III-J | 0.23 ± 0.02 | x | 0.25 | 1.38 | 0.082 | x |
P7-WT | ND | _ | ND | NA | NA | _ |
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Lakhssassi, N.; Baharlouei, A.; Meksem, J.; Hamilton-Brehm, S.D.; Lightfoot, D.A.; Meksem, K.; Liang, Y. EMS-Induced Mutagenesis of Clostridium carboxidivorans for Increased Atmospheric CO2 Reduction Efficiency and Solvent Production. Microorganisms 2020, 8, 1239. https://doi.org/10.3390/microorganisms8081239
Lakhssassi N, Baharlouei A, Meksem J, Hamilton-Brehm SD, Lightfoot DA, Meksem K, Liang Y. EMS-Induced Mutagenesis of Clostridium carboxidivorans for Increased Atmospheric CO2 Reduction Efficiency and Solvent Production. Microorganisms. 2020; 8(8):1239. https://doi.org/10.3390/microorganisms8081239
Chicago/Turabian StyleLakhssassi, Naoufal, Azam Baharlouei, Jonas Meksem, Scott D. Hamilton-Brehm, David A. Lightfoot, Khalid Meksem, and Yanna Liang. 2020. "EMS-Induced Mutagenesis of Clostridium carboxidivorans for Increased Atmospheric CO2 Reduction Efficiency and Solvent Production" Microorganisms 8, no. 8: 1239. https://doi.org/10.3390/microorganisms8081239