Affinity purification and characterization of 2,4-dichlorophenol hydroxylase from Pseudomonas cepacia

Radjendirane, V.; Bhat, Manzoor A.; Vaidyanathan, C.S.

doi:10.1016/0003-9861(91)90180-q

Cited by 37 publications

(31 citation statements)

References 28 publications

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“…In addition, there are no significant loss of enzyme activity when TfdB-JLU was stored for 50 days at 4°C. Under these conditions, the relative activity was above 90%, similar to that of the TfdB from Pseudomonas cepacia (Radjendirane et al 1991), which is stable and can be stored at 4°C for about 30 days without any appreciable loss of activity.…”

Section: Effects Of Temperature On Tfdb-jlu Activity and Stabilitymentioning

confidence: 60%

“…Although many chlorophenol hydroxylases have been isolated from different genera, only a few TfdB enzymes have been purified and characterised (Beadle and Smith 1982;Liu and Chapman 1984;Radjendirane et al 1991;Makdessi and Lechner 1997;Ledger et al 2006;Huong et al 2007). Their substrate specificities are similar, and most of them display high activity toward 2,4-DCP and 4-chloro-2-methylphenol.…”

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confidence: 99%

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Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil

Zhou

et al. 2011

Biotechnol Lett

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A novel 2,4-dichlorophenol hydroxylase (TfdB, EC 1.14.13.20) gene, designated as tfdB-JLU, was identified from a metagenome constructed from polychlorinated biphenyl-contaminated soil by functional screening and heterologously expressed in Escherichia coli. The deduced amino acid sequence of tfdB-JLU exhibited less than 48% homology with other known TfdBs. The enzyme exhibited a wider substrate spectrum than the previously reported TfdBs and higher relative activity towards ortho-substituted dichlorophenols, 2-chlorophenol, and 3-chlorophenol than towards 2,4-dichlorophenol, the preferred substrate of other known TfdBs. The enzyme had a K ( m ) of 5 μM for 2,4-dichlorophenol and 6 μM for NADPH. The optimal temperature and pH of the enzyme were 25°C and 7.5, respectively. Activity of the purified TfdB-JLU was slightly enhanced by Ca(2+), Mn(2+), Co(2+), and Fe(2+), and completely inhibited by Cu(2+), Hg(2+), and Zn(2+). This study is the first report to identify a novel TfdB from a metagenome.

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Section: Effects Of Temperature On Tfdb-jlu Activity and Stabilitymentioning

confidence: 60%

mentioning

confidence: 99%

Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil

Zhou

et al. 2011

Biotechnol Lett

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“…This is an indication for the relatedness of dehalogenating and nondehalogenating monooxygenases. This group of flavoproteins which all depend on the same cosubstrates, namely, flavin, O 2 , and NAD(P)H, have also the subunit size of about 60 kDa in common (3,11,19,29,34). They catalyze the hydroxylation of an aromatic substrate either ortho or para relative to the existing hydroxyl group.…”

Section: Discussionmentioning

confidence: 99%

Purification and characterization of 2,4,6-trichlorophenol-4-monooxygenase, a dehalogenating enzyme from Azotobacter sp. strain GP1

et al. 1997

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The enzyme which catalyzes the dehalogenation of 2,4,6-trichlorophenol (TCP) was purified to apparent homogeneity from an extract of TCP-induced cells of Azotobacter sp. strain GP1. The initial step of TCP degradation in this bacterium is inducible by TCP; no activity was found in succinate-grown cells or in phenol-induced cells. NADH, flavin adenine dinucleotide, and O 2 are required as cofactors. As reaction products, 2,6-dichlorohydroquinone and Cl ؊ ions were identified. Studies of the stoichiometry revealed the consumption of 2 mol of NADH plus 1 mol of O 2 per mol of TCP and the formation of 1 mol of Cl ؊ ions. No evidence for membrane association or for a multicomponent system was obtained. Molecular masses of 240 kDa for the native enzyme and 60 kDa for the subunit were determined, indicating a homotetrameric structure. Cross-linking studies with dimethylsuberimidate were consistent with this finding. TCP was the best substrate for 2,4,6-trichlorophenol-4-monooxygenase (TCP-4-monooxygenase). The majority of other chlorophenols converted by the enzyme bear a chloro substituent in the 4-position. 2,6-Dichlorophenol, also accepted as a substrate, was hydroxylated in the 4-position to 2,6-dichlorohydroquinone in a nondehalogenating reaction. NADH and O 2 were consumed by the pure enzyme also in the absence of TCP with simultaneous production of H 2 O 2 . The NH 2 -terminal amino acid sequence of TCP-4-monooxygenase from Azotobacter sp. strain GP1 revealed complete identity with the nucleotide-derived sequence from the analogous enzyme from Pseudomonas pickettii and a high degree of homology with two nondehalogenating monooxygenases. The similarity in enzyme properties and the possible evolutionary relatedness of dehalogenating and nondehalogenating monooxygenases are discussed.Polychlorinated phenols are widespread environmental pollutants, and their degradation has been the subject of numerous investigations (for reviews, see references 5, 9, 10, and 21). While many different microorganisms with the capacity to degrade these compounds have been isolated, relatively few reports have dealt with the initial step of their degradation. Moreover, the majority of these investigations were performed with whole cells, crude extracts, or partially purified enzymes, and so far only two polychlorophenol-dehalogenating enzymes from Burkholderia (formerly Pseudomonas) cepacia AC1100 (33) and from a Flavobacterium sp. (34) have been purified to homogeneity. Other polychlorophenol dehalogenases were partially purified from an Arthrobacter sp. (23) or found to be membrane-associated enzymes in Rhodococcus chlorophenolicus (26,27) and Mycobacterium fortuitum (28).The dehalogenation of 2,4,6-trichlorophenol (TCP) was detected in crude extracts of Streptomyces rochei 303 (7) and Pseudomonas pickettii (12). The conversion of TCP to 2,6-dichlorohydroquinone was also observed with whole cells from Pseudomonas cepacia AC1100 (25).In this paper, we describe the purification and characterization of a TCP-dehalogenating enzyme from t...

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“…Increasing Hg concentration to 0.4 mg/L leads to decrease in caffeine degradation by 27%, which can be classified as detrimental. Radjendirane et al, (1991) reported that Hg could also inhibit hydroxylases activity like aryl 2,4-dichlorophenol hydroxylase and 3-hydroxybenzoate-6-hydroxylase. Analysis of variance showed that there was no significant difference in caffeine degradation between 0.4, 0.5 0.6 and 0.7 mg/L (p˃0.05) but the remaining concentration shows there was a significant difference between them (p˂0.05).…”

Section: Bajopas Volume 8 Number 2 December 2015mentioning

confidence: 99%

Studies of action of heavy metals on caffeine degradation by immobilised <i>Leifsonia sp.</i> strain SIU

Ibrahim¹,

Muhammad²,

Tanko³

et al. 2016

Bayero J. Pure App. Sci.

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Caffeine is an important naturally occurring compound that can be degraded by bacteria. Excessive caffeine consumption is known to have some adverse problems. Previously, Leifsonia sp. strain SIU capable of degrading caffeine was isolated from agricultural soil. The bacterium was tested for its ability to degrade caffeine as the sole carbon and nitrogen source. The isolate was encapsulated in gellan gum and its ability to degrade caffeine in the presence of heavy metals was determined. Out of the nine heavy metals tested, Copper (Cu), Mercury (Hg), and Silver (Ag) had significant effects on caffeine degradation at 1mg/L. Therefore, the concentration of these heavy metals was varied from 0 -1 mg/L to see at what concentration each metal it has effect. Ag and Hg showed effect at 0.1 mg/L with caffeine degradation of 64.05 and 52.17% respectively, while Cu showed effect at 0.8 mg/L with caffeine degradation of 64.74%. These bacterium features make it an ultimate means for caffeine bioremediation. This is the first report of effect of heavy metals on caffeine degradation by immobilised Leifsonia sp. strain SIU.

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Affinity purification and characterization of 2,4-dichlorophenol hydroxylase from Pseudomonas cepacia

Cited by 37 publications

References 28 publications

Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil

Cloning and characterisation of a novel 2,4-dichlorophenol hydroxylase from a metagenomic library derived from polychlorinated biphenyl-contaminated soil

Purification and characterization of 2,4,6-trichlorophenol-4-monooxygenase, a dehalogenating enzyme from Azotobacter sp. strain GP1

Studies of action of heavy metals on caffeine degradation by immobilised <i>Leifsonia sp.</i> strain SIU

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