Isolation, Identification and Characterization of Bioflocculant-Producing Bacteria from Activated Sludge of Vulindlela Wastewater Treatment Plant
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolation of Bioflocculant-Producing Microorganisms
2.2. Screening of Bacteria for Bioflocculant Production
2.3. Determination of Bioflocculant Activity
2.4. Identification of a Bioflocculant-Producing Bacterium
2.4.1. Morphological Identification of the Bioflocculant-Producing Bacterium
2.4.2. Molecular Identification of Bioflocculant–Producing Bacterium
2.5. Activation of the Isolate for Fermentation
2.6. Optimisation of Bioflocculant Production Conditions
2.6.1. Determination of the Inoculation Volume
2.6.2. Effect of Carbon and Nitrogen Sources on Bioflocculating Activity
2.6.3. Effect of Metal Ions on Bioflocculating Activity
2.6.4. Effect of Initial pH of the Production Media
2.6.5. Effect of Shaking Speed on Bioflocculant Production
2.6.6. Effect of Cultivation Temperature on Bioflocculating Activity
2.6.7. Time Course Assay
2.7. Extraction and Purification of Bioflocculant
2.8. Chemical Analysis of the Purified Bioflocculant
2.8.1. Composition Analysis of the Purified Bioflocculant
2.8.2. FTIR Analysis
2.8.3. Thermo-Gravimetric Analysis (TGA) of a Bioflocculant
2.8.4. Elementary Analysis
2.9. Flocculation Characteristics of a Purified Bioflocculant
2.9.1. Effect of Dosage Concentration on Flocculating Activity
2.9.2. Effect of pH on the Flocculating Activity of Purified Bioflocculant
2.9.3. Effect of Metal Ions on Flocculating Activity
2.9.4. Effect of Heat on Flocculating Activity
2.10. Statistical Analysis
3. Results and Discussion
3.1. Isolation, Screening and Identification of Bioflocculant-Producing Bacterium
3.2. Optimisation of Culture Conditions
3.2.1. Effect of Inoculation Volume on Bioflocculant Production
3.2.2. Effect of Carbon Source on Biofloccculant Production
3.2.3. Effect of Nitrogen Sources on Bioflocculant Production
3.2.4. Effect of Metal Ions on Bioflocculant Production
3.2.5. Effect of pH on Bioflocculant Production
3.2.6. Effect of Shaking Speed on Bioflocculant Production
3.2.7. Effect of Temperature on Bioflocculant Production
3.2.8. Time Course on Bioflocculant Production
3.3. Extraction and Purification of a Bioflocculant
3.4. Chemical composition of the Purified Bioflocculant
3.5. Fourier Transform Infrared Spectroscopy (FTIR) Analysis
3.6. Thermogravimetric TGA Analysis of the Purified Bioflocculant
3.7. Elemental Analysis of the Purified Bioflocculant
3.8. SEM Analysis of the Purified Bioflocculant
3.9. Conditions for Flocculating Activity of the Purified Bioflocculant
3.9.1. Effect of Dosage Concentration on the Purified Bioflocculant
3.9.2. Effect of pH on the Purified Bioflocculant
3.9.3. The Effective Use of Metal Ions on the Purified Bioflocculant
3.9.4. Effect of Heat on the Purified Bioflocculant
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Ingredients | Amount (g) |
---|---|
Glucose | 20.0 |
K2HPO4 | 5.0 |
K2PO4 | 2.0 |
Urea | 0.5 |
Yeast extract | 0.5 |
MgSO4·7H2O | 0.2 |
(NH4)2SO4 | 0.2 |
NaCl | 0.1 |
Activated sludge water (filtered) | 1 Litre |
Sample | Un-Diluted (CFU/mL) | 10−1 (CFU/mL) | 10−2 (CFU/mL) |
---|---|---|---|
Wastewater | TN | TN | 8 × 103 |
Inoculation Volume (%) | FA (%) ± SD | Carbon Sources | FA (%) ± SD | Nitrogen Source | FA (%) ± SD | Cations | FA (%) ± SD |
---|---|---|---|---|---|---|---|
1 | 80.5 ± 0.41 a | Lactose | 52.7 ± 0.92 a | Casein | 52.4 ± 2.48 a | Control | 46.2 ± 0.13 a |
2 | 89.2 ± 0.14 b | Galactose | 56.1 ± 0.98 a | Ammonium sulphate | 64.3 ± 1.32 b | Li+ | 65.7 ± 1.15 b |
3 | 90.2 ± 0.14 c | Xylose | 63.9 ± 1.34 b | Ammonium acetate | 66.9 ± 0.17 b,c | K+ | 66.7 ± 0.05 b |
4 | 63.3 ± 0.15 d | Maltose | 71.7 ± 4.45 c | Ammonium chloride | 77.2 ± 0.57 d | Na+ | 86.1 ± 1.86 c |
5 | 49.4 ± 0.04 e | Fructose | 83.4 ± 2.59 d | Urea | 90.1 ± 2.39 e | Ba2+ | 90.4 ± 0.08 d |
- | - | Starch | 83.8 ± 0.99 d,e | Yeast extract | 91.4 ± 0.41 e,f | Ca2+ | 90.8 ± 0.02 d,e |
- | - | Glucose | 89.3 ± 1.68 d,f | Peptone | 92.1 ± 0.11 e,g | Mg2+ | 93.1 ± 0.02 e,f |
- | - | Sucrose | 94.7 ± 3.10 e | Mixture of nitrogen | 63.1 ± 0.44 b,h | Mn2+ | 95.1 ± 0.25 f,g |
- | - | Mixture (CHO) | 68.9 ± 1. 3 b,c,h | - | - | Fe3+ | 21.1 ± 1.01 h |
Sample | Concentration (%) |
---|---|
Total sugar | 64.25% |
Total protein | 10.42% |
Uronic acid | 23.51% |
Dosage (mg/mL) | FA (%) ± SD | pH | FA (%) ± SD | FA (%) ± SD | Metal Ions |
---|---|---|---|---|---|
0.2 | 57.1 ± 5.78 a | 3 | 81.1 ± 0.07 a | Na+ | 63.3 ± 0.15 a |
0.4 | 81.9 ±1.96 b | 4 | 86.4 ± 1.35 b | Li+ | 65.3 ± 0.13 a |
0.6 | 80.8 ± 6.31 b,c | 5 | 88.9 ± 0.48 c | K+ | 64.0 ± 0.21 a |
0.8 | 65.5 ± 1.75 a | 6 | 97.4 ± 0.05 d | Ca2+ | 70.2 ± 0.11 b |
1.0 | 49.4 ± 1.62 a,d | 7 | 93.3 ± 0.32 e | Ba2+ | 77.2 ± 0.42 c |
- | - | 8 | 80.2 ± 0.23 a | Mn2+ | 98.4 ± 0.13 d |
- | - | 9 | 78.2 ± 0.14 a,f | Mg2+ | 98.3 ± 0.14 d |
- | - | 10 | 65.6 ± 1.86 g | Fe3+ | 90.2 ± 0.16 e |
- | - | 11 | 60.4 ± 0.58 h | Control | 43.3 ± 0.10 f |
- | - | 12 | 63.7 ± 0.45 g,i | - | - |
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Nkosi, N.C.; Basson, A.K.; Ntombela, Z.G.; Maliehe, T.S.; Pullabhotla, R.V.S.R. Isolation, Identification and Characterization of Bioflocculant-Producing Bacteria from Activated Sludge of Vulindlela Wastewater Treatment Plant. Appl. Microbiol. 2021, 1, 586-606. https://doi.org/10.3390/applmicrobiol1030038
Nkosi NC, Basson AK, Ntombela ZG, Maliehe TS, Pullabhotla RVSR. Isolation, Identification and Characterization of Bioflocculant-Producing Bacteria from Activated Sludge of Vulindlela Wastewater Treatment Plant. Applied Microbiology. 2021; 1(3):586-606. https://doi.org/10.3390/applmicrobiol1030038
Chicago/Turabian StyleNkosi, Nkanyiso Celukuthula, Albertus K. Basson, Zuzingcebo G. Ntombela, Tsolanku S. Maliehe, and Rajasekhar V. S. R. Pullabhotla. 2021. "Isolation, Identification and Characterization of Bioflocculant-Producing Bacteria from Activated Sludge of Vulindlela Wastewater Treatment Plant" Applied Microbiology 1, no. 3: 586-606. https://doi.org/10.3390/applmicrobiol1030038