Triclosan (2, 4, 4'-trichloro-2'-hydroxyl diphenyl ether) is a broad-spectrum antimicrobial agent present in a number of house hold consumables. Aerobic and anaerobic enrichment cultures tolerating triclosan were developed and 77 bacterial strains tolerating triclosan at different levels were isolated from different inoculum sources. Biodegradation of triclosan under aerobic, anoxic (denitrifying and sulphate reducing conditions), and anaerobic conditions was studied in batch cultures with isolated pure strains and enrichment consortium developed. Under aerobic conditions, the isolated strains tolerated triclosan up to 1 g/L and degraded the compound in inorganic-mineral-broth and agar media. At 10 mg/L level triclosan, 95 ± 1.2% was degraded in 5 days, producing phenol, catechol and 2, 4-dichlorophenol as the degradation products. The strains were able to metabolize triclosan and its degradation products in the presence of monooxygenase inhibitor 1-pentyne. Under anoxic/anaerobic conditions highest degradation (87%) was observed in methanogenic system with acetate as co-substrate and phenol, catechol, and 2, 4-dichlorophenol were among the products. Three of the isolated strains tolerating 1 g/L triclosan were identified as Pseudomonas sp. (BDC 1, 2, and 3).
An isolated Serratia marcescens strain exhibited growth-coupled perchlorate (ClO4 -) reduction under anoxic conditions. Perchlorate was reduced completely with stoichiometric chloride buildup and equimolar acetate consumption. Polymerase chain reaction confirmed the presence of pcrA and cld genes coding for key enzymes involved in the ClO4 - degradation pathway. The isolate degraded ClO4 - under high salt (up to 15% NaCl) and a wide range of pH (4.0-9.0), as well as simultaneously reduced nitrate and ClO4 -.
Kinetics of chlorite dismutase (CD), the terminal enzyme involved in the perchlorate (ClO4(-)) reduction pathway, in a ClO4(-)-degrading bioreactor are reported in this study. Enzyme activity was determined from dissolved oxygen released during disproportionation of chlorite (ClO2(-)). CD activity was in the range 29.8-36.4 U/mg dry weight sludge, and kinetic constants Vmax and K(m) of the enzyme were 37.83 U/mg dry weight and 0.28 mM, respectively. Among reactor operational conditions, enzyme activity was observed at pH 4.0-9.0, with an optimum at pH 6.0. Redox potential in the range -50 to +120mV and NaCl up to 3.5 g/L had no significant effect on CD activity. However, co-occurring pollutants such as ammonium at 10 ppm, nitrite at 50 ppm and EDTA at 100 microM reduced CD activity substantially. The present study highlights ideal bioreactor conditions to avoid ClO2(-) toxicity, while indicating the buffering potential of a mixed microbial system against inhibiting factors to maintain stable CD activity in bioreactors.
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