Chloroform (CF) is largely produced by both anthropogenic and natural sources. It is detected in ground and surface water sources and it represents the most abundant halocarbon in the atmosphere. Microbial CF degradation occurs under both aerobic and anaerobic conditions. Apart from a few reports describing the utilization of CF as a terminal electron acceptor during growth, CF degradation was mainly reported as a cometabolic process. CF aerobic cometabolism is supported by growth on short-chain alkanes (i.e., methane, propane, butane, and hexane), aromatic hydrocarbons (i.e., toluene and phenol), and ammonia via the activity of monooxygenases (MOs) operatively divided into different families. The main factors affecting CF cometabolism are (1) the inhibition of CF degradation exerted by the growth substrate, (2) the need for reductant supply to maintain MO activity, and (3) the toxicity of CF degradation products. Under anaerobic conditions, CF degradation was mainly associated to the activity of methanogens, although some examples of CF-degrading sulfate-reducing, fermenting, and acetogenic bacteria are reported in the literature. Higher CF toxicity levels and lower degradation rates were shown by anaerobic systems in comparison to the aerobic ones. Applied physiological and genetic aspects of microbial cometabolism of CF will be presented along with bioremediation perspectives.
The ability of a Rhodococcus aetherovorans strain, BCP1, to grow on butane and to degrade chloroform in the 0-633 microM range (0-75.5 mg l(-1)) via aerobic cometabolism was investigated by means of resting-cell assays. BCP1 degraded chloroform with a complete mineralization of the organic Cl. The resulting butane and chloroform maximum specific degradation rates were equal to 118 and 22 micromol mg(protein)(-1)day(-1), respectively. Butane inhibition on chloroform degradation was satisfactorily interpreted by means of a model of competitive inhibition, with an inhibition constant equal to 38 % of the estimated butane half-saturation constant, whereas chloroform (at 11 microM) did not inhibit butane utilization. Acetylene (1,720 microM) induced an almost complete inactivation of the degradation of both butane and chloroform, indicating that the studied cometabolic process is mediated by a monooxygenase enzyme. BCP1 proved capable of degrading vinyl chloride and 1,1,2-trichloroethane, but not 1,2-trans-dichloroethylene. BCP1 could grow on the intermediates of the most common butane metabolic pathways and on the aliphatic hydrocarbons from ethane to n-heptane. After growth on n-hexane, it was able to deplete chloroform (13 microM) with a degradation rate higher than that obtained, at the same chloroform concentration, after growth on butane.
Rhodococcus sp. strain BCP1, known for its capacity to grow on short-chain n-alkanes (C 2 to C 7 ) and to cometabolize chlorinated solvents, was found to also utilize medium-and long-chain n-alkanes (C 12 to C 24 ) as energy and carbon sources. To examine this feature in detail, a chromosomal region which includes the alkB gene cluster encoding a non-heme di-iron monooxygenase (alkB), two rubredoxins, and one rubredoxin reductase was cloned from the BCP1 genome. Furthermore, the activity of the alkB gene promoter (P alkB ) was examined in the presence of gaseous, liquid, and solid n-alkanes along with intermediates of the putative n-alkane degradation pathway. A recombinant plasmid, pTP alkB LacZ, was constructed by inserting the lacZ gene downstream of P alkB , and it was used to transform Rhodococcus sp. strain BCP1. Measurements of -galactosidase activity showed that P alkB is induced by C 6 to C 22 n-alkanes. Conversely, C 2 to C 5 and >C 22 n-alkanes and alkenes, such as hexene, were not inducers of alkB expression. The effects on P alkB expression induced by alternative carbon sources along with putative products of n-hexane metabolism were also evaluated. This report highlights the great versatility of Rhodococcus sp. strain BCP1 and defines for the first time the alkB gene transcriptional start site and the alkB promoter-inducing capacities for substrates different from n-alkanes in a Rhodococcus strain.
The goals of this work were (i) to compare two anion ion exchange resins (IRA958 Cl and IRA67) and a nonionic resin (XAD16) in terms of phenolic compounds adsorption capacity from olive mill wastewater and (ii) to compare the adsorption capacity of the best resin on columns of different length. The ion exchange resins performed worse than nonionic XAD16 in terms of resin utilization efficiency (20% versus 43%) and phenolic compounds/COD enrichment factor (1.0 versus 2.5). The addition of volatile fatty acids did not hinder phenolic compounds adsorption on either resin, suggesting a noncompetitive adsorption mechanism. A pH increase from 4.9 to 7.2 did not affect the result of this comparison. For the best performing resin (XAD16), an increase in column length from 0.5 to 1.8 m determined an increase in resin utilization efficiency (from 12% to 43%), resin productivity (from 3.4 to 7.6 g sorbed phenolics /kg resin ), and phenolics/COD enrichment factor (from 1.2 to 2.5). An axial dispersion model with nonequilibrium adsorption accurately interpreted the phenolic compounds and COD experimental curves.
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