Jatinder K. Bewtra

Removal of phenolic compounds from synthetic wastewater using soybean peroxidase

Abstract Experiments were conducted to investigate the efficiency of using soybean peroxidase (SBP) to remove several different phenolic compounds from unbuffered synthetic wastewater. The phenol derivatives studied included parent phenol, chlorinated phenols, cresols, 2,4-dichlorophenol and 4,4′-isopropylidenediphenol (commonly known as bisphenol A). Optimum conditions to achieve at least 95% removal of these compounds were determined for the following parameters: pH, SBP dose in the absence and presence of polyethylene glycol (PEG), hydrogen peroxide to substrate ratio, and PEG dose. Experimental results showed that SBP efficiently removed aromatic compounds from synthetic wastewater in the presence of hydrogen peroxide. An increase in the hydrogen peroxide to substrate ratio beyond the optimum resulted in enzyme inactivation in all cases except for bisphenol A. The optimum pH for different phenolic compounds ranged from 5.5 to 8. For each substrate, the optimum enzyme dose in the presence of PEG varied significantly. The studies showed that PEG only slightly reduced the amount of SBP required for 95% removal of the substrate. For most of the substrates, an increase in PEG dose beyond the optimum dose did not significantly change the removal efficiency.

Inactivation of horseradish peroxidase by phenol and hydrogen peroxide: a kinetic investigation

Inactivation of horseradish peroxidase (HRP) was examined in the presence of hydrogen peroxide alone and in the presence of hydrogen peroxide plus phenol. HRP is inactivated upon exposure to hydrogen peroxide (H2O2) by the combination of two possible pathways, dependent upon hydrogen peroxide concentration. At low H2O2 concentrations (below 1.0 mM in the absence of phenol), inactivation is predominantly reversible, resulting from the formation and accumulation of catalytically inert intermediate compound III. As H2O2 concentrations increase, an irreversible mechanism-based inactivation process becomes predominant. The overall inactivation comprised of both processes exhibits a second-order inactivation rate constant (kapp) of 0.023 +/- 0.005 M-1 s-1 at pH 7.4 and 25 degrees C. In the presence of both hydrogen peroxide fixed at 0.5 mM and phenol, HRP was inactivated in an irreversible, time- and phenol concentration-dependent process, also mechanism-based, with a kapp of 0.019 +/- 0.004 M-1 s-1.

A model for the Protective Effect of Additives on the Activity of Horseradish Peroxidase in the Removal of Phenol

Horseradish peroxidase has been proven effective in removing phenolic compounds in wastewater and additives such as polyethylene glycol have been found very effective in reducing the minimum enzyme dose required. The effect of additives on horseradish peroxidase-catalyzed removal of phenol was investigated in this study. In the absence of additive, active enzyme is predominantly inactivated by the polymer product formed during the reaction. The specific activity of horseradish peroxidase is higher due to the presence of additive. Experiments suggest that additives combine with the polymerization products formed during the reaction, because additives have a higher partition affinity with the polymer products than peroxidases. Most of the polymer product is coupled with additive so that less enzyme interacts with the polymer product. Horseradish peroxidase still combines with polymer products and becomes inactivated but at a much slower rate when additives are present. Consequently, the enzyme activity is protected by the additives.

Phenol conversion and dimeric intermediates in horseradish peroxidase-catalyzed phenol removal from water.

published in Advance ACS Abstracts, September 1, 1994. 2180 Environ. Scl. Technol., Vol. 28, No. 12, 1994

Optimization of the reaction conditions for enzymatic removal of phenol from wastewater in the presence of polyethylene glycol

Abstract The effect of the additive, polyethylene glycol (PEG), on the horseradish peroxidase (HRP) catalysed removal of phenol from wastewater has been studied over the phenol concentration range of 1–10 mM (0.1–1.0 g/l). The optimum pH, HRP concentration, PEG concentration and the molar ratio of hydrogen peroxide and phenol have been investigated in the presence of PEG at room temperature in order to achieve the maximum phenol removal efficiency with the minimum cost. The effect of concentrations of HRP and PEG on reaction time was also investigated. Experimental results showed that the addition of PEG had significant protective effect on the activity of HRP. The amount of peroxidase required was reduced 40- and 75-fold less than that required without PEG for 1 and 10 mM phenol solutions, respectively. The higher the phenol concentration, the more effective was the addition of PEG. In the presence of PEG, the optimum pH is 8.0 and the optimum molar ratio of hydrogen peroxide and phenol is around 1.0. The minimum doses of HRP and PEG required for at least 95% removal were determined for several phenol concentrations and two empirical models are proposed to predict the minimum HRP and PEG doses required for 95% removal over the entire phenol concentration range of 1–10 mM. Under the optimum reaction conditions described above, the reaction times for at least 95% removal from 1 and 10 mM phenol solutions were 5 and 3 h, respectively. An increase in HRP concentration significantly reduced the reaction time; however, an increase in PEG concentration showed negligible influence.

REACTOR DEVELOPMENT FOR PEROXIDASE CATALYZED POLYMERIZATION AND PRECIPITATION OF PHENOLS FROM WASTEWATER

Abstract Horseradish peroxidase enzyme catalyzes the oxidation of toxic aromatic compounds, especially phenols and aromatic amines, in the presence of hydrogen peroxide. The reaction products polymerize to form high molecular weight materials which readily precipitate from solution; hence, providing a means for the treatment of wastewaters which contain aromatic compounds. The catalytic lifetime of horseradish peroxidase enzyme can be extended by optimizing treatment conditions such as pH and temperature and by maintaining a low instantaneous enzyme concentration in the reaction mixture. The enzyme catalyzed polymerization process was implemented in a continuous stirred tank reactor (CSTR) configuration because reactant and enzyme concentrations are lowered immediately upon entering the reactor causing a reduction in inactivation of HRP through free radical bonding and compound III formation. Catalytic turnovers achieved in single and multiple CSTRs in series were significantly higher than those observed in batch reactors when sufficient retention time was provided.

Comparison of additives in the removal of phenolic compounds by peroxidase-catalyzed polymerization

Several additives, polyethylene glycol, gelatin and some polyelectrolytes, were studied to compare their behaviour in the removal of phenolic compounds from aqueous solution by horseradish-peroxidase-catalyzed polymerization. The effects of additives on optimum pH range, horseradish-peroxidase saving, reaction stoichiometry and minimum additive requirements were investigated. The fate of additive after reaction was also studied. The experimental results showed that all tested additives significantly reduced the horseradish-peroxidase requirement. Comparison among them revealed that polyethylene glycol was the best choice from all perspectives. It saved more peroxidase and had no negative overdose effect shown by gelatin and polyelectrolytes. At minimum polyethylene glycol dose, there was little polyethylene glycol remaining in solution after completion of reaction. However, a considerable amount of gelatin remained in solution even at the minimum gelatin dose. Also, gelatin produced more precipitate than polyethylene glycol.

Sulfide Production by Sulfate Reducing Bacteria with Lactate as Feed in an Upflow Anaerobic Fixed Film Reactor

The sulfate reducing bacteria (SRB) have the capability of reducing sulfate (SO4-2) under anaerobic conditions into sulfide (S-2) which can precipitate metals as metal sulfides. The optimum conditions for sulfide production by SRB utilizing lactate, in an upflow anaerobic fixed film reactor (UAFFR) were not previously established. The main objective of this research was to investigate these conditions for the growth of SRB to ensure the highest sulfide production under consistent behaviour of the system. Substrate containing lactate as the organic carbon source along with sulfate, nitrogen and phosphorus as the required nutrients was used as a feed to the UAFER which was seeded with SRB. It was found that an optimum sulfide production occurred with an organic loading rate (OLR) of 6 kg d-1 m-3, while the theoretical oxygen demand to sulfate ratio (ThOD/SO4) ranged from 1.5 to 2.25. Also, the optimum total nitrogen and phosphorus demands were determined to be about 250 and 50 mg L-1 respectively. A total nitrogen concentration above 600 mg L-1 started showing toxicity and lowered the sulfide production. The optimum ThOD:N:P for sulfide production and growth of SRB in the UAFFR was 100:5:1 under optimum conditions.

Effect of temperature on oxygen transfer in water

Abstract The effect of temperature on oxygen transfer by diffused-air aeration is presented. The study was made in a full scale aeration tank using both saran tubes and spargers as the diffusion media at aeration rates from 7 to 32 ft 3 /min per foot of tank length. The oxygen transfer was measured in water through a temperature range 10–30°C in 2.5°C increments. The results are analyzed statistically and are summarized in the form of a mathematical expression.

Immobilized enzyme catalyzed removal of 4-chlorophenol from aqueous solution

Abstract The ability of horseradish peroxidase enzyme attached on three different reactor matrices: cellulose filter paper, nylon balls and nylon tubing, to remove 4-chlorophenol from aqueous solution is evaluated. The enzymatic reaction is extremely fast and the reaction products remained on the reactor matrix. Detachment or release of the reaction products from the reactor matrix is not observed. Results indicate that, over 80% removal efficiency can be obtained as long as enzyme activity is not limiting in the reactor. Systematic recycle batch reactor studies reveal that, initial reaction rates exhibit saturation with substrate concentration under conditions of excess peroxide. Inactivation of enzyme active sites by reaction intermediates is observed in the reactor studies. Immobilized peroxidase also catalyzes the oxidation of other chlorophenols and cresols.