Types Of Oxygenases Research Articles

Assembling a genome for novel nitrogen-fixing bacteria with capabilities for utilization of aromatic hydrocarbons

Metagenome from refinery wastewater treatment plant running under nitrogen stress was analyzed for mining of novel aromatic hydrocarbon-degrading bacteria. The sequence data were assembled using metaspade followed by binning using the Metabat tool to assemble genome; where coverage and depth were calculated using bowtie and samtools. The analysis picked a novel genome belonging to family Bradyrhizobiaceae, identified based on 16S rDNA gene which was supported by CheckM and Kraken analysis. Using RAST, the assembled genome showed the capabilities for nitrogen fixation with the utilization of multiple hydrocarbon substrates with 14 different types of oxygenases as mapped by Minpath. An additional genetic feature like genes for stress and resistance towards heavy metals and antibiotic suggested that the genome has gone through the rigorous process of adaptation. If such bacteria could be cultivated then it will open the broad window of bioremediation strategies under nitrogen stress environment.

A Comprehensive Review of Aliphatic Hydrocarbon Biodegradation by Bacteria.

Hydrocarbons are relatively recalcitrant compounds and are classified as high-priority pollutants. However, these compounds are slowly degraded by a large variety of microorganisms. Bacteria are able to degrade aliphatic saturated and unsaturated hydrocarbons via both aerobic and anaerobic pathways. Branched hydrocarbons and cyclic hydrocarbons are also degraded by bacteria. The aerobic bacteria use different types of oxygenases, including monooxygenase, cytochrome-dependent oxygenase and dioxygenase, to insert one or two atoms of oxygen into their targets. Anaerobic bacteria, on the other hand, employ a variety of simple organic and inorganic molecules, including sulphate, nitrate, carbonate and metals, for hydrocarbon oxidation.

Differential Expression of Cytochrome P450 Genes Regulate the Level of Adipose Arachidonic Acid in Sus Scrofa

We compared the fatty acid composition of adipose tissues prepared from Korean native and Yorkshire pigs that have different characteristics in growth and fat deposition. There was no significant difference in the content of most fatty acids between the two breeds, with the exception of arachidonic acid and cis-11,14,17-eicosatrienoic acid. We also investigated the transcriptional levels of genes encoding three different types of oxygenases, including cytochrome P450 (CYP), lipoxygenase and cyclooxygenase, which metabolize arachidonic acid. We found a significant difference in the expression of the CYP genes, CYP2A13, CYP2U1 and CYP3A4, but no differences for the latter two genes between the two breeds. Our results suggest that the difference in arachidonic acid content between the two breeds was caused by differential expression of the CYP genes. Eventually, different levels of EETs and HETEs produced from arachidonic acid by the activity of CYP might contribute partly to the difference of fatness between the two breeds. (Key Words : Arachidonic Acid, Cytochrome P450, Gene Expression, Adipose Tissue, Sus scrofa)

A medium‐throughput screening assay to determine catalytic activities of oxygen‐consuming enzymes: a new tool for functional characterization of cytochrome P450 and other oxygenases

A challenge of the post-genomic era is to determine the functions of a plethora of orphan genes. This is a more acute problem when dealing with large gene families, such as the superfamily encoding cytochrome P450 enzymes in higher plants. We propose here a new, simple, medium-throughput methodology to screen for potential substrates of orphan P450 mono-oxygenases. The same technique can also be applied to screening for inhibitors of the oxygenases involved in the biosynthesis of compounds essential for plant development, such as growth regulators. The method is based on a commercially available microplate system, which detects the oxygen consumed by the catalytic reaction via an oxygen-sensing fluorophore. It is optimized using as a model CYP73A1, the cinnamic acid hydroxylase from Helianthus tuberosus, expressed in yeast. We show that the procedure is suitable not only for the detection and real-time monitoring, but also for the quantitative evaluation of enzyme activity. This new method has broad application for the identification of candidate substrates and inhibitors in chemical libraries, to support determination of physiological substrates, development of plant growth regulators, investigations on herbicide and pollutant metabolism, synthesis of valuable compounds and drug design. It also provides a fast-assay platform for determination of catalytic and inhibition parameters. The method applies to plant P450 enzymes, but also to cytochromes P450 from other organisms, and all types of oxygenases. The critical steps, calculation of oxygen consumption from fluorescence signal, and limits of the methods are discussed.