Towards a sustainable bioprocess for the remediation of acid mine drainage
- Authors: Mambo, Mutsa Prudence
- Date: 2011
- Subjects: Acid mine drainage , Algae culture , Reduction (Chemistry) , Hydrolysis , ASPAM model (Acid mine drainage) , Water -- Purification
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:5955 , http://hdl.handle.net/10962/d1006167 , Acid mine drainage , Algae culture , Reduction (Chemistry) , Hydrolysis , ASPAM model (Acid mine drainage) , Water -- Purification
- Description: Acid mine drainage is of growing concern for both developing and developed economies. Thus there is increasing pressure to develop alternative remediation strategies. Biological sulphidogenic mechanisms have long since been studied but, very few have been implemented on a large scale. Limitations are due to the inability to acquire a suitable, low cost, environmentally friendly, renewable carbon source. The present study investigated the use of an algae biomass generated by the HRAOP of an IAPS as a carbon source for the EBRU 00AB/06 SRB consortium. The algae biomass and consortium were utilized together to remediate simulated AMD. Remediation involved decreasing the sulphate and metal concentrations in solution and decreasing the acidity of a simulated AMD. Experiments were carried out to investigate the capability of the EBRU 00AB/06 SRB consortium for sulphate reduction and sulphide generation. The consortium produced colonies when grown under anaerobic conditions in Petri dishes containing modified lactate SRB medium. The SRB consortium reduced the sulphate concentration of modified Postgates medium B and generated sulphide. Further analysis of the EBRU 00AB/06 SRB consortium revealed that the consortium was minimally impacted at pH 5 and by sulphate and iron at 3 g.L-1 and 0.5 g.L-1 respectively. The EBRU 00AB/06 SRB consortium was exposed to Actinomycin D and Ethidium Bromide to determine whether transcription and translation of proteins was required for sulphate reduction. Results indicated that sulphide generation and sulphate reduction were inducible. Analysis of the algae biomass used in this study revealed the empirical formula C1.0H1.91N0.084S0.003O0.36 indicating a carbon source rich in the nutrients required to sustain microbial development. Light microscopy revealed that algae cell walls and in particular those of Pediastrum were susceptible to acid hydrolysis. Dinitrosalicylic acid, Nile red, Bradford and Ninhydrin assays were used to determine the reducing sugar, lipid, protein and amino acid content respectively, of the mixed algae biomass. Results showed that upon exposure of the biomass to simulated AMD at pH 1 and pH 3, the concentration of reducing sugars and amino acids in solution increased. Whereas levels of lipids remained unchanged while the protein concentration decreased, indicating that, upon exposure of algae biomass to AMD, simulated or otherwise, cells ruptured, proteins were hydrolyzed and polysaccharides were broken down to sugars which are immediately available for SRB utilization. Exposure of biomass to simulated AMD revealed further that the presence of algae biomass increased the pH of simulated AMD (pH 3) to pH 7.67 after 4 d. Likewise, the pH of simulated AMD at 1 increased to 1.77 after 2 d while pH of the neutral control increased to 8.1 after 4 d. A direct comparison between lactate and algae biomass revealed 94 % sulphate removal after 23 d in the presence of algae biomass while 82 % sulphate removal was measured in the presence of lactate. Thus the EBRU 00AB/06 SRB consortium successfully utilized algae biomass for sulphate reduction and sulphide generation. In another experiment to establish if the consortium could remediate simulated AMD (pH 5) containing 0.5 g.L-1 iron and 3 g.L-1 sulphate while utilizing an algae biomass as the carbon source no residual iron was detected after 14 d and by day 23, an 89.07 % reduction in sulphate was measured. The results of this investigation are discussed in terms of utilizing a readily available and renewable biomass in the form of microalgae produced in HRAOPs as an effective carbon source in the SRB catalysed remediation of AMD.
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- Date Issued: 2011
The bioaccumulation of platinum (IV) from aqueous solution using sulphate reducing bacteria: role of a hydrogenase enzyme
- Authors: Rashamuse, Konanani Justice
- Date: 2003
- Subjects: Sulfur bacteria , Bioremediation , Enzymes -- Metabolism , Platinum , Platinum compounds , Reduction (Chemistry) , Hydrogenation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4003 , http://hdl.handle.net/10962/d1004063 , Sulfur bacteria , Bioremediation , Enzymes -- Metabolism , Platinum , Platinum compounds , Reduction (Chemistry) , Hydrogenation
- Description: The enzymatic reduction of a high-valence form of metals to a low-valence reduced form has been proposed as a strategy to treat water contaminated with a range of metals and radionuclides. Metal reduction by sulphate reducing bacteria (SRB) is carried out either chemically (involving reduction by hydrogen sulphide) or enzymatically (involving redox enzymes such as the hydrogenases). While reduction of metal ions by hydrogen sulphide is well known, the enzymatic mechanism for metal reduction is poorly understood. The aims of this study were to investigate the role of SRB in facilitating platinum removal, and to investigate the role of a hydrogenase in platinum reduction in vitro. In order to avoid precipitation of platinum as platinum sulphide, a resting (non-growing) mixed SRB culture was used. The maximum initial concentration of platinum (IV), which SRB can effectively remove from solution was shown to be 50 mg.l⁻¹. Electron donor studies showed high platinum (IV) uptake in the presence of hydrogen, suggesting that platinum (IV) uptake from solution by SRB requires careful optimization with respect to the correct electron donor. Transmission electron microscopy (TEM) and energy dispersive X-ray (EDX) analysis indicated that platinum was being precipitated in the periplasm, a major area of hydrogenase activity in SRB. Purification of the hydrogenase by ammonium sulphate precipitation (65%), Toyopearl-Super Q 650S ion exchange and Sephacry 1 S-100 size exclusion chromatography revealed that the hydrogenase was monomeric with a molecular weight of 58 KDa, when analyzed by 12% SDS-PAGE. The purified hydrogenase showed optimal temperature and pH at 35°C and 7.5 respectively, and a poor thermal stability. In vitro investigation of platinum reduction by purified hydrogenase from mixed SRB culture showed that hydrogenase reduces platinum only in the presence of hydrogen. Major platinum (IV) reduction was observed when hydrogenase was incubated with cytochrome C₃ (physiological electron carrier in vivo) under hydrogen. The same observations were also noted with industrial effluent. Collectively these findings suggest that in vitro platinum reduction is mediated by hydrogenase with a concerted action of cytochrome C₃ required to shuttle the electron from hydrogenase.
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- Date Issued: 2003