Enzymes Involved in Naproxen Degradation by Planococcus sp. S5

Abstract

A b s t r a c tNaproxen is a one of the most popular non-steroidal anti-inflammatory drugs (NSAIDs) entering the environment as a result of high consumption. For this reason, there is an emerging need to recognize mechanisms of its degradation and enzymes engaged in this process. Planococcus sp. S5 is a gram positive strain able to degrade naproxen in monosubstrate culture (27%). However, naproxen is not a sufficient growth substrate for this strain. In the presence of benzoate, 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid or vanillic acid as growth substrates, the degradation of 21.5%, 71.71%, 14.75% and 8.16% of naproxen was observed respectively. It was shown that the activity of monooxygenase, hydroxyquinol 1,2-dioxygenase, protocatechuate 3,4-dioxygenase and protocatechuate 4,5-dioxyegnase in strain S5 was induced after growth of the strain with naproxen and 4-hydroxybenzoate. Moreover, in the presence of naproxen activity of gentisate 1,2-dioxygenase, enzyme engaged in 4-hydroxybenzoate metabolism, was completely inhibited. The obtained results suggest that monooxygenase and hydroxyquinol 1,2-dioxygenase are the main enzymes in naproxen degradation by Planococcus sp. S5. 178acid produced by Prunus domestica, Melissa officinalis or Vitis vinifera or vanillic acid identified in Juglans regia or Chenopodium murales (Ghareib et al., 2010;Kakkar and Bais, 2014). The objective of our study is to examine the ability of Planococcus sp. S5 to degrade naproxen in either mono-or disubstrate cultures with plant aromatic compounds as growth substrates. Plano coccus sp. S5 was chosen for this study since it degrades salicylate (Łabużek et al., 2003), which is not only the intermediate of naphthalene degradation and mimetic of naproxen, but also belongs to non-steroidal antiinflammatory drugs. Additionally, the aim of this study is to identify enzymes engaged in naproxen degradation by Planococcus sp. S5 in cometabolic conditions. Experimental Materials and MethodsMedia and culture conditions. Planococcus sp. S5 was routinely cultivated in BBL nutrient broth at 30°C and 130 rpm for 24 hours. Then 6 mg/l naproxen was added to the culture. After 48 hours, cells were harvested by centrifugation (5.000 × g at 4°C for 15 min), washed with fresh sterile medium and used as inoculum. Degradation of naproxen in a monosubstrate, as well as cometabolic systems, was performed in 500 ml Erlenmeyer flasks containing 250 ml of the mineral salts medium (Greń et al., 2010) inoculated with cells to a final optical density of about 1.5 and 0.5 at λ = 600 nm (OD600) for the monosubstrate and cometabolic systems, respectively. For degradation experiments two control cultures were prepared. The uninoculated control (I) consisted of 250 ml of sterile mineral salts medium, while the heatkilled control (II) consisted of 250 ml of autoclaved culture prepared under conditions identical to those of the experimental cultures. Naproxen was added to each flask to obtain a final concentration of 6 mg/l, and all cultures were incubated with shakin...

Naproxen in the environment: its occurrence, toxicity to nontarget organisms and biodegradation

“…Hydroxyquinol 1,2dioxygenase is involved in the cleavage of naproxen in Stenotrophomonas maltophilia KB2 and Planococcus sp. S5 (Wojcieszyńska et al 2014;Wojcieszyńska et al 2016 proposed the transformation of naproxen by eliminating the methyl group. The O-desmethylnaproxen that is f o r m e d m a y c o n j u g a t e w i t h t y r o s i n e t o 6 , 7dihydroxynaphthalene-2-yl-tyrosine.…”

Immobilization of Planococcus sp. S5 Strain on the Loofah Sponge and Its Application in Naproxen Removal

“…Therefore, naproxen has been releasing into the natural environment in an unchanged form for over 40 years. Due to its continuous accumulation in the environment, naproxen is now one of the most frequently detected drug in surface and drinking water (concentration in the range 0.01-2.6 µg/L) [1][2][3].…”

Effect of Pseudomonas moorei KB4 Cells’ Immobilisation on Their Degradation Potential and Tolerance towards Paracetamol

“…The following initial optical density settings were tested: 0.2, 0.4, 0.6, 0.8, 1, 1.2 and 1.4). Flasks were incubated under shaking conditions (130 rpm) at different temperature settings (20,25,30,35 and 40 • C) and for different time intervals (24, 48 and 72 h) to find the optimal conditions. Parameters were optimised in the following order: time, pH, initial OD600 and temperature.…”

“…Pollutants such as NSAIDs (in parent form or as their metabolites, often more active and/or stable) are, however, incessantly being released into the environment for reasons such as the lack of optimal treatment techniques [6]. Bioremediation of contaminated groundwater or soil is considered to be a relatively environmentally friendly, [7][8][9][10][11][12][13][14][15][16][17][18][19][20]. P. moorei KB4 is one of the few described bacterial strains capable of degrading either paracetamol (4-hydroxyacetanilide, N-acetyl-p-aminophenol, acetaminophen), an analgesic and antipyretic drug often described as an NSAID, or diclofenac, an NSAID generally accepted as being dangerous for the environment [21].…”