Investigation of Acetoin Biosynthesis by Bacillus subtilis ACA-DC 1176 Growing on Crude Glycerol in Flask and Bioreactor Trials
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microorganism and Media
2.2. Cultures of B. subtilis
2.2.1. Preculture
2.2.2. Shake-Flask and Duran Bottle Cultures
2.2.3. Bioreactor Culture
2.3. Analytical Methods
2.3.1. Determination of Physicochemical Characteristics of Raw Materials
2.3.2. Growth Determination
2.3.3. Substrate and Metabolites Quantitative Determination
2.3.4. Estimation of kLa Value
2.4. Data Analysis
3. Results
3.1. Initial Trials on Acetoin Production Feasibility
3.2. Investigation of Acetoin Production by B. subtilis ACA-DC 1176 in Shake-Flask Cultures
3.2.1. Culture Medium and Temperature
3.2.2. Feeding Strategy
3.3. Scale-Up in Bioreactor Culture
4. Concluding Remarks
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Oxygen Regime | Carbon Source | B. subtilis ACA-DC 1176 | B. subtilis ACA-DC 1225 | ||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
Acetoin (g/L) | BDO (g/L) | DCW (g/L) | SubC (%) | Time (h) | Acetoin (g/L) | BDO (g/L) | DCW (g/L) | SubC (%) | Time (h) | ||
Aerobiosis | |||||||||||
Glucose | 3.8 | 3.3 | 3.4 | 100 | 45 | 4.9 | 2.5 | 3.8 | 100 | 48 | |
Glycerol | 3.5 | 1.2 | 5.7 | 100 | 44 | 2.5 | 1.3 | 5.8 | 100 | 55 | |
Crude glycerol | 2.6 | 1.3 | 5.0 | 100 | 45 | 2.4 | 0.8 | 4.8 | 100 | 70 | |
Anaerobiosis | |||||||||||
Glycerol | - | - | 1.1 | 12 | - | Neg | - | 3.8 | 50 | 70 | |
Crude glycerol | Neg | - | 1.1 | 15 | 70 | - | - | Neg | - | - |
No. | Gly Type | Nitrogen | Temp | Gly0 | Acetoin | BDO | DCW | YACTN/Gly | ACTN/BDO | ConRate | Time | ||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Total | ComOrg | CSL | (°C) | (g/L) | (g/L) | (g/L) | (g/L) | (g/g) | (g/g) | (g/L/h) | (h) | ||
1 | Pu. | 100% | 100% | - | 30 | 43.3 | 9.0 | 4.4 | 12.7 | 0.21 | 9.0 | 0.50 | 83 |
2 | 67.0 | 17.5 | 6.1 | 14.3 | 0.27 | 6.3 | 0.54 | 116 | |||||
3 | Pu. | 100% | 50% | 50% | 30 | 40.6 | 11.2 | 4.5 | 6.9 | 0.28 | 7.8 | 0.57 | 93 |
4 | 61.9 | 5.6 | 13.8 | 5.1 | 0.12 | 0.4 | 0.49 | 135 | |||||
5 | Pu. | 50% | 50% | - | 30 | 41.0 | 2.0 | 2.2 | 5.2 | Neg | 1.1 | 0.27 | 157 |
6 | 62.5 | 1.9 | 0.9 | 4.2 | Neg | 1.5 | 0.37 | 184 | |||||
7 | Cr. | 100% | 100% | - | 30 | 38.3 | 8.5 | 4.2 | 10.3 | 0.22 | 8.0 | 0.51 | 81 |
8 | 60.1 | 10.1 | 6.9 | 11.0 | 0.17 | 7.1 | 0.56 | 128 | |||||
9 | Cr. | 100% | 50% | 50% | 30 | 42.8 | 8.0 | 3.9 | 13.1 | 0.19 | 8.0 | 0.54 | 85 |
10 | 62.4 | 5.8 | 6.7 | 14.0 | 0.11 | 0.9 | 0.48 | 136 | |||||
11 | Cr. | 100% | 50% | 50% | 37 | 44.0 | 11.2 | 6.5 | 7.4 | 0.25 | 1.0 | 0.49 | 90 |
12 | 63.9 | 12.5 | 7.5 | 9.6 | 0.20 | 1.5 | 0.49 | 120 | |||||
13 | Cr. | 50% | 50% | - | 37 | 41.0 | 1.7 | 3.3 | 4.4 | Neg | 0.9 | 0.31 | 145 |
14 | 62.5 | 2.0 | 3.7 | 5.6 | Neg | 0.5 | 0.42 | 150 |
Time Periods (h) | kLa (1/h) | ConRate (g/L/h) | ACTN (g/L) | BDO (g/L) | DCW (g/L) | ACTN/BDO (g/g) | ||||
---|---|---|---|---|---|---|---|---|---|---|
Δt | t | Δ | Total | Δ | Total | |||||
1 | 0–70.5 | 64.7 | 54.1 | 0.80 | +7.4 | 7.4 | +7.8 | 7.8 | 10.4 | 0.94 |
70.5 | 70.5 | 1.20 | +5.9 | 13.3 | −4.7 | 3.1 | 10.6 | 4.29 | ||
2 | 70.5–108.7 | 94.5 | 87.3 | 1.00 | +9.8 | 23.1 | +4.9 | 8.0 | 10.4 | 2.22 |
108.7 | 103.3 | 1.30 | +1.4 | 24.5 | +6.2 | 14.2 | 10.5 | 1.73 | ||
3 | 108.7–137 | 130.2 | 103.3 | 0.90 | +3.4 | 27.9 | +3.5 | 17.7 | 10.0 | 1.58 |
137 | 87.3 | 1.00 | +4.2 | 32.1 | +5.2 | 22.9 | 10.4 | 1.40 | ||
4 | 137–183.7 | 167.7 | 87.3 | 0.80 | +2.2 | 34.3 | +4 | 26.9 | 10.9 | 1.28 |
183.7 | 0.20 | +1.7 | 36.0 | −1.4 | 25.5 | 10.5 | 1.41 | |||
Sum | 183.7 | 0.88 |
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Karayannis, D.; Moraiti, E.; Kalantzi, O.; Papanikolaou, S. Investigation of Acetoin Biosynthesis by Bacillus subtilis ACA-DC 1176 Growing on Crude Glycerol in Flask and Bioreactor Trials. Reactions 2024, 5, 664-679. https://doi.org/10.3390/reactions5040034
Karayannis D, Moraiti E, Kalantzi O, Papanikolaou S. Investigation of Acetoin Biosynthesis by Bacillus subtilis ACA-DC 1176 Growing on Crude Glycerol in Flask and Bioreactor Trials. Reactions. 2024; 5(4):664-679. https://doi.org/10.3390/reactions5040034
Chicago/Turabian StyleKarayannis, Dimitris, Eleni Moraiti, Ourania Kalantzi, and Seraphim Papanikolaou. 2024. "Investigation of Acetoin Biosynthesis by Bacillus subtilis ACA-DC 1176 Growing on Crude Glycerol in Flask and Bioreactor Trials" Reactions 5, no. 4: 664-679. https://doi.org/10.3390/reactions5040034