Analysis of Two Gene Clusters Involved in the Degradation of 4-Fluorophenol by
            <i>Arthrobacter</i>
            sp. Strain IF1

Ferreira, Maria Isabel M.; Iida, Toshiya; Hasan, Syed A.; Nakamura, Kaoru; Fraaije, Marco W.; Janssen, Dick B.; Kudo, Toshiaki

doi:10.1128/aem.00171-09

Cited by 27 publications

(26 citation statements)

References 58 publications

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“…To investigate the possible role of such a monooxygenase in the conversion of 2-FP, we performed experiments with cell-free extracts prepared from cells grown on 2-FP, with addition of purified FAD reductase (FpdB) from the 4-fluorophenol-degrading bacterium Arthrobacter sp. strain IF1 (Ferreira et al 2009). Conversion was followed by HPLC analysis, which showed formation of 3-FC from 2-FP.…”

Section: Identification and Isolation Of A Monooxygenase Involved In mentioning

confidence: 98%

“…Bacterial conversion of several chlorophenols and 4-fluorophenol can proceed by two-component monooxygenases that use NADPH as electron donor (Louie et al 2002;Nordin et al 2005;Ferreira et al 2008Ferreira et al , 2009). The reduced nicotinamide cofactor is used by the reductase component Fig.…”

Section: Identification and Isolation Of A Monooxygenase Involved In mentioning

confidence: 99%

“…Monooxygenase activity was measured in 50 mM Tris-Cl buffer (pH 7.5) in reaction mixtures containing 1 mM ascorbate, 5 μM FAD, 3 μg of flavin reductase (FpdB, purified as described by Ferreira et al (2009)), 180 U ml −1 catalase (from bovine liver, Fluka), 1.2 mM NADH, and 600 μM of 2-FP. Incubations were carried out at 25°C.…”

Section: Enzyme Assaysmentioning

confidence: 99%

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Isolation and characterization of a Rhodococcus strain able to degrade 2-fluorophenol

Duque

Hasan

Bessa

et al. 2011

Appl Microbiol Biotechnol

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A pure bacterial culture able to utilize 2-fluorophenol (2-FP) as sole carbon and energy source was isolated by selective enrichment from sediments collected from a contaminated site in Northern Portugal. 16S rRNA gene analysis showed that the organism (strain FP1) belongs to the genus Rhodococcus. When grown aerobically on 2-FP, growth kinetics of strain FP1 followed the Luong model. An inhibitory effect of increasing 2-FP concentrations was observed with no growth occurring at 2-FP levels higher than ca. 4 mM. Rhodococcus strain FP1 was able to degrade a range of other organofluorine compounds, including 2-fluorobenzoate, 3-fluorobenzoate, 4-fluorobenzoate, 3-fluorophenol, 4-fluorophenol, 3-fluorocatechol, and 4-fluorocatechol, as well as chlorinated compounds such as 2-chlorophenol and 4-chlorophenol. Experiments with cell-free extracts and partially purified enzymes indicated that the first step of 2-fluorophenol metabolism was conversion to 3-fluorocatechol, suggesting an unusual pathway for fluoroaromatic metabolism. To our knowledge, this is the first time that utilization of 2-FP as a growth substrate by a pure bacterial culture is reported.

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Section: Identification and Isolation Of A Monooxygenase Involved In mentioning

confidence: 98%

Section: Identification and Isolation Of A Monooxygenase Involved In mentioning

confidence: 99%

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Isolation and characterization of a Rhodococcus strain able to degrade 2-fluorophenol

Duque

Hasan

Bessa

et al. 2011

Appl Microbiol Biotechnol

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confidence: 99%

“…Most studies on the bacterial degradation of fluorinated organics describe fluorobenzoic acids, which under aerobic conditions can be converted into the corresponding fluorocatechols (3, 17, 33). Papers about the degradation of fluorophenols have also appeared (11,25,51).4-Fluorocinnamic acid (4-FCA) is used in industry for the synthesis of flavors and pharmaceuticals (8), and polymers of 4-FCA are applied in electronics (14). It was proposed that under aerobic conditions in nonacclimated industrial activated sludge, 4-FCA could be converted into 4-fluorobenzoic acid (4-FBA) via the formation of 4-fluoroacetophenone (4-FAP) (8, 32).…”

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confidence: 99%

Complete Biodegradation of 4-Fluorocinnamic Acid by a Consortium Comprising Arthrobacter sp. Strain G1 and Ralstonia sp. Strain H1

Hasan

Ferreira

Koetsier

et al. 2011

Appl Environ Microbiol

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Fluorinated organic compounds are of growing industrial importance, with applications such as agrochemicals, pharmaceuticals, and performance materials (23,24,28,41). The safe use of such compounds, as well as appropriate disposal and treatment of wastes, will benefit from knowledge about their biodegradation. However, little information is available about the microbial metabolism of fluorinated organic compounds compared to other halogenated chemicals. Most studies on the bacterial degradation of fluorinated organics describe fluorobenzoic acids, which under aerobic conditions can be converted into the corresponding fluorocatechols (3, 17, 33). Papers about the degradation of fluorophenols have also appeared (11,25,51).4-Fluorocinnamic acid (4-FCA) is used in industry for the synthesis of flavors and pharmaceuticals (8), and polymers of 4-FCA are applied in electronics (14). It was proposed that under aerobic conditions in nonacclimated industrial activated sludge, 4-FCA could be converted into 4-fluorobenzoic acid (4-FBA) via the formation of 4-fluoroacetophenone (4-FAP) (8, 32). In another study, using activated sludge from a wastewater treatment plant, 4-FCA was suggested to be transformed into an epoxide that is converted to 4-FAP. This compound would then be converted into 4-FBA, but no products were detected from the breakdown of 4-FBA (5).The conversion of nonhalogenated cinnamic acids to benzoic acids, such as the transformation of ferulic acid to vanillic acid, has been described previously (2,4,20,31,39). Cinnamic acid, coumaric acid, and ferulic acid are transformed by Streptomyces setonii (47) and Rhodopseudomonas palustris to benzoic acid or the corresponding derivatives (19). Alcanivorax borkumensis MBIC 4326 (9) and Papillibacter cinnamivorans (7) transformed cinnamic acid into benzoic acid. The metabolism of these compounds thus proceeds with side chain degradation prior to ring cleavage. Side chain degradation is carried out either by ␤-oxidation or by direct deacetylation mechanisms, which leads to elimination of two carbon units from the unsaturated side chain in bacteria, yeasts, and fungi (40).Since no clear information on the degradation route of 4-FCA is available, we have isolated two pure bacterial strains to study the complete microbial metabolism of the compound, and in this paper, we propose a degradation pathway. MATERIALS AND METHODSGrowth conditions. Cells of strains G1 and H1 were grown aerobically at 30°C under rotary shaking or in a fermentor. Growth medium (MMY) contained (per liter) 5.37 g of Na 2 HPO 4 ⅐ 12H 2 O, 1.36 g of KH 2 PO 4 , 0.5 g of (NH 4 ) 2 SO 4 , and 0. Enrichment and isolation of 4-FCA-and 4-FBA-degrading organisms. Soil samples collected from a site in the Netherlands contaminated mainly with chlorobenzene and halogenated aliphatic compounds were used as the initial inocula for the 4-FCA and 4-FBA enrichment cultures. Flasks contained 40 ml MMY and 5 mM 4-FCA or 5 mM 4-FBA as the sole source of carbon and energy. The cultures were incubated at room temperatur...

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Identification of a Hotspot Residue for Improving the Thermostability of a Flavin‐Dependent Monooxygenase

et al. 2019

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HadA is a flavin‐dependent monooxygenase that can catalyze the denitration and dehalogenation of a wide variety of toxicants such as pesticides. Although these enzymatic reactions are useful for bioremediation or biocatalysis, the application of HadA for these purposes is not yet possible because of its low thermostability. In this work we have engineered HadA to be more thermostable through the use of structural, in silico, and rational approaches. The X‐ray structure of HadA was solved to obtain a reliable three‐dimensional protein model for further prediction of thermostable variants. In silico analysis by using two bioinformatic tools—FireProt and Disulfide by Design—suggested 102 variants that we then further refined by applying rational criteria including the location of a particular residue and its nearby interactions, as well as other biophysical parameters to narrow down the list to six candidates. The G513Y variant was found to be an optimal engineered candidate because it has significantly improved stability relative to the wild‐type enzyme and equivalent activity. G513Y has an activity half‐life 72 (50 °C) and 160 times (45 °C) longer than that of the wild‐type enzyme. Coupled together with thermostable reactions of reduced flavin and NADH‐regenerating systems, the G513Y variant can be used to catalyze denitration of 4nitrophenol at 45 °C. Structure/sequence alignments of HadA and its homologues indicate that several flavin‐dependent monooxygenases also contain amino acid residues homologous to the G513 of HadA, hence opening up the possibility of applying this engineering approach to improving their thermostabilities as well. Molecular dynamics (MD) simulations confirmed that the improved thermostability of the G513Y variant was due to aromatic hydrocarbon interactions between Y513 and N359, L347, G348, and F349.

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Analysis of Two Gene Clusters Involved in the Degradation of 4-Fluorophenol by Arthrobacter sp. Strain IF1

Cited by 27 publications

References 58 publications

Isolation and characterization of a Rhodococcus strain able to degrade 2-fluorophenol

Isolation and characterization of a Rhodococcus strain able to degrade 2-fluorophenol

Complete Biodegradation of 4-Fluorocinnamic Acid by a Consortium Comprising Arthrobacter sp. Strain G1 and Ralstonia sp. Strain H1

Identification of a Hotspot Residue for Improving the Thermostability of a Flavin‐Dependent Monooxygenase

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