This pathway of styrene degradation is of potential biotechnological relevance for the production of especially phenylacetic acids. Pseudomonas putida ca3 is capable of converting the aromatic hydrocarbon styrene, its metabolite phenylacetic acid, and glucose into polyhydroxyalkanoate pha when a limiting concentration of nitrogen as sodium ammonium phosphate is supplied to the growth medium. Styrene, which is widely used in manufacturing, is both acutely and chronically toxic to mice. This report describes the isolation, sequencing and analysis of a new chromosomal fragment deriving from. Construction of a recombinant biocatalyst for the production. Accumulation of polyhydroxyalkanoate from styrene and.
Furthermore, antibioticsonthe basis of penicillin can be obtained from 4hydroxyphenylacetic acid or nonsubstituted phenylacetic acid 9,11. Bruker dmx 400mhz data collected by madison metabolomics consortium. Although a phenylacetate metabolic gene cluster had been identified, the. Catalytical oxidation of styrene by molecularly imprinted. Phenylacetic acid benzoic acid no reaction substitution elimination oxidation 1b. The acid was then taken into 10% nahco 3 solution and was washed with ethyl acetate 3x30 ml, separated the organic layer, and the aqueous layer was acidified with dilute hcl, to yield the pure phenylacetic acid as solid. Figures uploaded by kevin edward o connor author content. Although a phenylacetate metabolic gene cluster had been identified. Research article open access regulation of phenylacetic acid. Finally, phenylacetaldehyde would be oxidized to become phenylacetic acid. Pseudomonas putida ca3 is capable of converting the aromatic hydrocarbon styrene, its metabolite phenylacetic acid, and glucose into polyhydroxyalkanoate.
The phenylacetic acid paa pathway is an important ring. Styrene lower catabolic pathway in pseudomonas fluorescens. Here, we focused on a simple enzymatic epoxidation of alkenes using lipase and phenylacetic acid. Ketoglutaric acid h mg b1 b2 b3 succinic acid h iron b2 malic acid h b3.
Transcriptional regulation of styrene degradation in. Phenylacetic acid is obtained by oxidation of phenylacetaldehyde with air or oxygen in. The cluster of styrene upper pathway catabolic genes sty genes has been previously localized on a chromosomal region. Apr 22, 2016 this pathway comprises a styrene monooxygenase smo, which oxidizes styrene to styrene oxide, a styrene oxide isomerase soi, which converts styrene oxide into phenylacetaldehyde, and a phenylacetaldehyde dehydrogenase pad. Davidson national high magnetic field laboratory, 1800 east paul dirac dr. Phenylacetic acid 4a and its esters are widely applied as flavors and fragrances. Pdf accumulation of polyhydroxyalkanoate from styrene and. In the illicit drug world, phenylacetic acid is often trafficked as a precursor for the bootleg manufacture of methamphetamine, and thus large purchases are monitored by drug enforcement agencies. Phenylacetic acid via the willgerodt reaction from styrene. This pathway comprises a styrene monooxygenase smo, which oxidizes styrene to styrene oxide, a styrene oxide isomerase soi, which converts styrene oxide into phenylacetaldehyde, and a phenylacetaldehyde dehydrogenase pad. Gc procedures have been described for the quantitative determination of urinary phenylglyoxylic and mandelic acids, which involve solvent extraction of the. The peroxy phenylacetic acid generated was then utilised directly for in situ oxidation of alkenes.
Pha accumulation occurs to a low level when the nitrogen concentration drops below 26. Styrene, the starting material for polystyrene synthesis, is a. Aug 10, 2010 aromatic compounds constitute the second most abundant class of organic substrates and environmental pollutants, a substantial part of which e. Us3919305a process for the preparation of phenylacetic acid. This method can be improved for pipelining continuous production, styrene.
Pha accumulation occurs to a low level when the nitrogen concentration drops. Styrene oxide is then isomerised into phenylacetaldehyde. Research article open access regulation of phenylacetic. As a result it is used as a perfumery and flavoring ingredient phenylacetic acid abr. This molecule is then able to be cleaved into acetyl coa which can enter the tca cycle. By the way, the reaction between phenylacetic acid and acetone is claimed to result in phenyl2propanone not bmk, my mistake, with the acetone being regarded as a precursor to acetic acid. Synthesizing phenylacetic acid from ethylbenzene, benzyl chloride, benzyl cyanide, mandelic acid, or acetophenone and styrene can be completed in the laboratory or at the industrial level. We previously made a link in burkholderia cenocepacia between pa degradation and. Cometabolic formation of substituted phenylacetic acids. It is reported here that expression of the regulatory genes stys and styr is essential for transcription of the upper pathway, but not for. The free phenylacetic acid is precipitated by the addition of conc hcl 3ml, in 92% yield, mp 7676. Microbial metabolism of styrene oligomers, 602 styrene is an important industrial chemical and it is also produced naturally, mainly by the decarboxylation of cin namic acid. As a commercial chemical, because it can be used in the illicit production of phenylacetone used in the. How to prepare phenylacetic acid from 2phenylethanol is to be stated.
Mandelic acid is an aromatic alpha hydroxy acid with the molecular formula c 6 h 5 chohco 2 h. In the lower pathway, phenylacetic acid will be joined to coa to make phenylacetylcoa by a ligase. Synthesizing phenylacetic acid from ethylbenzene, benzyl chloride, benzyl cyanide, mandelic acid, or acetophenone and styrene can be completed in the. Research article open access regulation of phenylacetic acid uptake is s54 dependent in pseudomonas putida ca3 niall d o leary1,2, mark m o mahony2 and alan dw dobson1,2 abstract background.
Bacterial phenylalanine and phenylacetate catabolic. I tried this once and had a yield of 7% actually, i did not even bother characterizing it, but it was a carboxylic acid and had the same typical smell as reagent grade phch2cooh. Process for the preparation of phenylacetic acid rhone. It is a metabolite of phenylalanine and is excreted in the urine of patients suffering from diseases like phenylketonuria. Methylglutaric acid h coq10 hmgcoa reductase inhibition. Citric acid cycle intermediates cisaconitic acid h isocitric acid h b3 mg mn. Pseudomonas fluorescens st is a styrene degrading microorganism that, by the sequential oxidation of the vinyl side chain, converts styrene to phenylacetic acid. The maximum yields for pha from styrene, phenylacetic acid, and glucose are 0. The latter enzyme enables the oxidation of the aldehyde to the central metabolite phenylacetic acid 1. Evidences of aromatic degradation dominantly via the. The purpose of these studies was to characterize the importance of this. It is a white crystalline solid that is soluble in water and polar organic solvents. Bacterial phenylalanine and phenylacetate catabolic pathway.
Because the structure of transition state is similar to that of complex of phenylacetic acid and hemin, we applied phenylacetic acid as the template to form the complex with hemin molecule which is comonomer and provides activity center for synthesized polymers. Styrene is metabolized by cytochromes p450 to the toxic metabolite styrene oxide, which is detoxified via hydrolysis with microsomal epoxide hydrolase meh playing a major role. Mar 22, 2018 this way efficiently transforms styrene into phenylacetic acid as central metabolite. In case of products from hydroxy acetophenones, the products were extracted into ethyl acetate. Phenylacetic acid is an important metabolite for several pathways and undergoes degradation during a subsequent catabolic route olivera et al. However, styrene is not a true xenobiotic and microbial pathways for its aerobic assimilation, via an intermediate, phenylacetic acid, have been identified in a diverse range of environmental isolates. Styrene degradation via phenylacetic acid was shown for the strains described. As a commercial chemical, because it can be used in. A number of soil bacteria is able to metabolize styrene via initial sidechain oxygenation into the central metabolite phenylacetic acid and harbors corresponding genes. Phenylacetic acid is used in some perfumes, possessing a honeylike odour in low concentrations, and is also used in penicillin g production. Jun 11, 2016 a number of soil bacteria is able to metabolize styrene via initial sidechain oxygenation into the central metabolite phenylacetic acid and harbors corresponding genes. The styrene degradative pathway in pseudmonas putida ca3 has previously been shown to be divided into an upper pathway involving the conversion of styrene to phenylacetic acid and a lower pathway for the subsequent degradation of phenylacetic acid. The potential for microbial bioremediation of styrene waste has received considerable research attention over the last number of years. Phenylacetic acid may arise from exposure to styrene plastic or through the consumption of fruits and vegetables.
Ppbrxryqalvlmvuhfffaoysan styrene chemical compound. Hydrolysis of chlorinated styrene a new route for the synthesis of phenylacetic acid and aethoxyphenylacetic acid. Styrene, sulfur, liquid ammonia and water into the autoclave, under 165. Because of the versatile applications of phenylacetic acids, different chemical strategies for their synthesis have been. Endogeneously, it is a catabolite of phenylalanine. Phenylacetic acid paa conjugate base phenylacetate, also known by various synonyms, is an organic compound containing a phenyl functional group and a carboxylic acid functional group.
Styrene degradation via phenylacetic acid was s hown for the strains described. It has a role as a human metabolite and a mouse metabolite. Pseudomonas putidas role in the bioremediation of plastic. Chemoenzymatic epoxidation of alkenes and reusability study.
Surprisingly, the bacterial catabolism of phenylalanine and phenylacetate remained an unsolved problem. The peroxy phenylacetic acid generated was then utilised directly for in situ oxidation of. Pdf accumulation of polyhydroxyalkanoate from styrene. Styrene r1, r2 h is transformed to phenylacetic acid through enzymes of sidechain oxygenation by the following steps.
Catabolism of phenylacetic acid in escherichia coli. Ohbutyric acid h balance of fat and cho metabolism. Escherichia coli living in the animal gut encounters aromatic compounds such as phenylacetic acid pa, 1 phenylpropionic acid, and their hydroxylated derivatives, as a result of the action of intestinal microflora on plant constituents, the amino acids phenylalanine and tyrosine, fatty acids with a terminal phenyl substituent, and some of their metabolites. The phenylacetic acid pa degradative pathway is the central pathway by which various aromatic compounds e. Chemoenzymatic epoxidation of alkenes and reusability. Sciencemadness discussion board p2p from recycled styrene.
Upper pathways for different aromatic compounds converge at common intermediate phenylacetylcoa pacoa, which is then metabolized to succinylcoa and acetylcoa. Cometabolic transformation of substituted styrenes was. Cometabolic formation of substituted phenylacetic acids by styrene. The cooled reaction mixture was taken up in chloroform some sulfur remained undissolved and the chloroform solution was washed successively with an equal volume of water.
Cro 3 in acidic solutions oxidize 1 0 alcohols directly into acids and 2 0 alcohols to ketones. How would you prepare the following compounds from 2. Jul 22, 2004 pseudomonas putida ca3 is capable of converting the aromatic hydrocarbon styrene, its metabolite phenylacetic acid, and glucose into polyhydroxyalkanoate pha when a limiting concentration of nitrogen as sodium ammonium phosphate is supplied to the growth medium. The willgerodt reaction can also be used to transform acetophenone or styrene to phenylacetic acid. The product 2phenyloxirane is also known as styrene oxide and can be converted by a styrene oxide isomerase soi to obtain phenylacetaldehyde, which can be transformed into the keyintermediate phenylacetic acid by a phenylacetaldehyde dehydrogenase pad. This way efficiently transforms styrene into phenylacetic acid as central metabolite. The immobilised candida antarctica lipase b, novozym 435 was used to catalyse the formation of peroxy acid instantly from hydrogen peroxide h 2 o 2 and phenylacetic acid. The paa cluster of escherichia coli w involved in the aerobic catabolism of phenylacetic acid pa has been cloned and sequenced.
Predicted data is generated using the us environmental protection agencys episuite. Accumulation of polyhydroxyalkanoate from styrene and phenylacetic acid by pseudomonas putida ca3. Cometabolic transformation of substituted styre nes was shown. Styrene lower catabolic pathway in pseudomonas fluorescens st. Cometabolic formation of substituted phenylacetic acids by. Microbial metabolism and biotransformations of styrene.
1180 300 918 254 361 345 182 1288 53 687 46 1494 1137 96 1322 1294 1284 457 923 1195 624 780 158 909 1215 1033 1051 576 1228 1254 763 845 1039 803 645 1446 148 979 160 615 1227