Cathodic reduction characteristics of 2-chloro-4-nitrophenol in microbial electrolysis cell

Microbial electrolysis cell (MEC) has been constructed to study the degradation characters of 2-chloro-4-nitrophenol (2C4NP) in waste water. The effects of applied voltage, initial concentration of substrate and co-matrix species on the reduction and degradation of 2C4NP were studied. Qualitative and quantitative analysis of 2C4NP residues and degradation intermediate by using UV-Vis, HPLC, HPLC/MS/MS, IC and other analytical testing techniques. The degradation mechanism of 2C4NP in MEC cathode was proposed. The results showed that electron and electroactive microorganisms would produce coupling effect and accelerate the degradation of 2C4NP under adding 0.5 V DC; Under the condition of satisfying the C/N ratio of electroactive anaerobic microorganism, the addition of organic substances such as glucose and sodium acetate which were easily degraded by microorganisms would hinder the degradation of 2C4NP in the cathode compartment. 2C4NP can be effectively degraded by adding appropriate amount of glucose as carbon source with the low C/N. 2C4NP undergoes reduction, dechlorination, denitrification and assimilation in the cathode compartment to form 2-chloro-4-aminophenol, 4-aminophenol, 2-chlorophenol, 2-chloro-4-hydroxyphenol, nitrophenol, hydroquinone, 4-hydroxyhexadienoic acid semialdehyde, valeric acid, oxalic acid and many other intermediate products. According to the degradation intermediates, the degradation mechanism of 2C4NP in the cathode compartment was presumed.

Expression of a Pseudomonas putidaAminotransferase Involved in Lysine Catabolism Is Induced in the Rhizosphere

ABSTRACT Using a transposon carrying a promoterless luxoperon to generate transcriptional fusions by insertional mutagenesis, we have identified a Pseudomonas putida gene with increased expression in the presence of corn root exudates. Expression of the transcriptional fusion, induced by the amino acid lysine, was detected in P. putida in the rhizosphere of plants as well as in response to seed exudates. The mutant was unable to grow on lysine or δ-aminovalerate as carbon sources, which indicates that the affected function is involved in the pathway for lysine catabolism. However, the mutant strain grew with glutaric acid, the product of δ-aminovalerate metabolism via glutaric acid semialdehyde, as a C source. The translated sequence of the interrupted gene showed high levels of similarity with aminotransferases. These sets of data suggest that the product of this gene has δ-aminovalerate aminotransferase activity. This is the first direct genetic evidence correlating a DNA sequence with such activity inPseudomonadaceae.

Degradation of 1,3-Dichloropropene by Pseudomonas cichorii 170

ABSTRACT The gram-negative bacterium Pseudomonas cichorii 170, isolated from soil that was repeatedly treated with the nematocide 1,3-dichloropropene, could utilize low concentrations of 1,3-dichloropropene as a sole carbon and energy source. Strain 170 was also able to grow on 3-chloroallyl alcohol, 3-chloroacrylic acid, and several 1-halo-n-alkanes. This organism produced at least three different dehalogenases: a hydrolytic haloalkane dehalogenase specific for haloalkanes and two 3-chloroacrylic acid dehalogenases, one specific for cis-3-chloroacrylic acid and the other specific for trans-3-chloroacrylic acid. The haloalkane dehalogenase and thetrans-3-chloroacrylic acid dehalogenase were expressed constitutively, whereas the cis-3-chloroacrylic acid dehalogenase was inducible. The presence of these enzymes indicates that 1,3-dichloropropene is hydrolyzed to 3-chloroallyl alcohol, which is oxidized in two steps to 3-chloroacrylic acid. The latter compound is then dehalogenated, probably forming malonic acid semialdehyde. The haloalkane dehalogenase gene, which is involved in the conversion of 1,3-dichloropropene to 3-chloroallyl alcohol, was cloned and sequenced, and this gene turned out to be identical to the previously studieddhaA gene of the gram-positive bacterium Rhodococcus rhodochrous NCIMB13064. Mutants resistant to the suicide substrate 1,2-dibromoethane lacked haloalkane dehalogenase activity and therefore could not utilize haloalkanes for growth. PCR analysis showed that these mutants had lost at least part of the dhaA gene.

Oxygenation and spontaneous deamination of 2-aminobenzenesulphonic acid in Alcaligenes sp. strain O-1 with subsequent meta ring cleavage and spontaneous desulphonation to 2-hydroxymuconic acid

2-Aminobenzenesulphonic acid (2AS) is degraded by Alcaligenes sp. strain O-1 via a previously detected but unidentified intermediate. A mutant of strain O-1 was found to excrete this intermediate, which was isolated and identified by m.s., 1H- and 13C-n.m.r. as 3-sulphocatechol (3SC). Proteins from cell extracts of strain O-1 were separated by anion-exchange chromatography. A multicomponent oxygenase was observed to convert 1 mol each of NADH, O2 and 2AS into 1 mol each of 3SC, NH3 and NAD+. The enzyme presumably catalysed formation of the ring of a 2-amino-2,3-diol moiety, and elimination in the amino group led to a rearomatization. 3SC was further degraded via meta ring cleavage, which could be prevented by inactivation of the 3-sulphocatechol-2,3-dioxygenase (3SC23O) with 3-chlorocatechol. In Tris buffer, the separated 3SC23O catalysed the reaction of 1 mol each of 3SC and O2 involving a transient yellow intermediate, and release of 1 mol of sulphite and two organic products. The major product was identified by n.m.r. and by g.c./m.s. as 5-carboxypenta-2,4-dien-5-olide (CPDO), an indicator of formation of 2-hydroxymuconic acid (2HM). The second product was identified as the Z,E isomer of 2HM by comparison with authentic material. When the CPDO in the product mixture was chemically hydrolysed to (Z,E)-2HM, 1 mol of (Z,E)-2HM/mol of 3SC was observed. If oxygenation of 3SC by 3SC23O was carried out in phosphate buffer, only a single product was detected, a keto form of 2HM. This dioate was also formed from authentic (Z,E)-2HM in phosphate buffer. Formation of the natural product (Z,E)-2HM from the xenobiotic, 3SC, seems to involve oxygenation to the unstable 2-hydroxy-6-sulphonomuconic acid semialdehyde, which hydrolyses spontaneously to 2HM. There would appear to be at least one spontaneous reaction per enzyme reaction in this pathway.

Oxidative Ring-Cleavage of Catechol in meta-Position by Superoxide

Superoxide reacts with catechol in dimethylsulfoxide to form a yellow product. The structure of this product was established by different methods. UV-spectra of the yellow compound corresponded under all circumstances to those of enzymatically produced 2-hydroxymuconic acid semialdehyde. In the presence of ammonia the product was converted to picolinic acid. The UV - spectra, the retention times in GC and HPLC and the mass spectra after methylation corresponded to those of authentic picolinic acid. Thus, the yellow reaction product was unequivocally identified as 2-hydroxymuconic acid semialdehyde. The formation of the me/a-cleavage product was only observed at low catechol concentrations (<2 mM) , at higher concentrations the product was not formed. The reaction was complete within 10 min at room temperature. Spectroscopic data suggest a first intermediate in the reaction with λmax = 804 nm, 772 nm and 660 nm.