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Physiology

X. azovorans are Gram-negative bacteria with cells 0. 5 to 1 μm in width and 1 to 3 μm in length. The organism is know as strain KF46F^T and was grown on nutrient agar for three days at 30°C.The carbon and energy source used for cultivation was carboxy-Orange II. Under the direction of a light microscope, the organism was found to give rise to circular, yellow-pigmented colonies. After cultivation, X. azovorans were determined to be aerobic, motile, and non-spore forming. X. azovorans grows at an optimal temperature of 30°C It is also important to note that strain KF46F^T is able to grow on various media like nutrient broth (30°C) and Luria-Burtani, but is usually not able to degrade carboxy-Orange II when grown on these media. Strain KF46F^T consists of predominant polar lipids such as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, and has an unknown aminophospholipid.

Genomics

The complete genome of X. azovorans DSM 13620^T has been sequenced by the DOE Joint Genome Institute (JGI). The bacteria has 6349 genes and 6280 protein coding genes.

The 16s ribosomal RNA gene of X. azovorans KF46F^T has been amplified using the polymerase chain reaction (PCR) and has been sequenced. The gene has a sequence length of 1484 base pairs. Researchers performed pulse field gel electrophoresis, a similar method described by Barton et al., and determined that the strain contains two plasmids of sizes 100 and 350 kb. Per high performance liquid chromatography (HPLC) methods described by Mesbah and Whitman, GC content of X. azovorans KF46F^T was determined to be approximately 70 percent.

Metabolism

X. azovorans is a chemoorganoheterotroph that carries out oxidative phosphorylation and uses oxygen as a terminal electron acceptor. The organism also has a gene predicted for nitrate reduction. The major quinone isolated was ubiquinone Q-8.This isolation was performed by HPLC methods as described by B.J. Tindall.

Based off of research performed by Blumel et al., the organism was characterized by growth on different carbon sources and sugar fermentation. The characterization methods were taken from Kampfer et al. The organism is able to use a number of amino acids, sugars, and carboxylic acids as a carbon and energy source.A few examples include utilization of D-Fructose and D-Mannitol. Based off of pathways shown on KEGG, 10.51% of X.azovoran's genome is comprised of genes contribute to amino acid metabolism. As far as carbohydrate metabolism is understood, the organism also has a complete TCA cycle and glycolysis pathway on KEGG. Around 6.79% of the organism's genes contribute to Xenobiotic biodegradation and metabolism. Specifically, the organism has genes predicted for aminobenzoate and benzoate degradation.

The organism tests positive for oxidase and catalase, but cannot produce urease, unlike its closely related neighbor Xenophilus aerolatus.

Ecology

X. azovorans was cultivated from the oral microbiota of domestic dogs. Researchers identified the bacterium by using comparative 16s rRNA sequencing. Specifically, a small percentage of cultivable X. azovorans was found in the dental plaque of the dogs.

X. azovorans has also been found in a compost-packed biofilter. The biofilter was treated with benzene-contaminated air. The bacterium was identified by using microbial population fingerprinting methods (differentiates microorganisms based on different characteristics) and the subsequent sequencing of fragments in the population by PCR. As the amount of benzene on the filter increased, the amount of cultivable bacteria increased as well. This was determined by cell plate counting and ribosomal intergenic spacer analysis (RISA).

Medical Relevance

Aerobic azoreductases make a significant contribution to the aerobic treatment of wastewaters which are colored by azo dyes. Azo dyes have been determined to be xenobiotic compounds that have characteristics that defer biodegradation. Due to this significant use, the azoreductase gene from X. azovorans strain KF46F^T was purified using affinity chromatography methods and cloned using PCR. Specifically, the gene has been determined to have high activity with the following azo dyes: Acid Orange 7, 1-(2-Pyridylazo)-2-naphthol, Solvent Orange 7, and Acid Red 88.Untreated wastewater can be harmful to human populations due to the role they play in mutagenic activity.Research was performed at an azo dye processing plant which is near a large river and a drinking-water treatment plant. It was found that 3% of waste from the azo dye processing plant ended up in the river that provides water to thousands of people. This is a very dangerous situation because it has been suggested that CYP450 enzymes in the human intestine activate azo dyes. Nevertheless, it has been determined that the intestine would suffer greatly due to this mutagenic activation as well as DNA in colon cells. Other studies, such as the one performed by Myslak et al., determined that painters exposed to azo dyes for a long period of time developed bladder cancer. All in all, it is important that more research be done on the X. azovorans azoreducatase gene due to its ability to break down chemicals in wastewater and to potentially prevent many humans from developing intestinal diseases.