Bacterial dioxygenase-catalysed dihydroxylation and chemical resolution routes to enantiopure cis-dihydrodiols of chrysene

Boyd, Derek R.; Sharma, Narain D.; Agarwal, Rakhi; Resnick, Sol M.; Schocken, Mark J.; Gibson, David T.; Sayer, Jane M.; Yagi, Haruhiko; Jerina, Donald M.

doi:10.1039/a606686k

Cited by 28 publications

(31 citation statements)

References 15 publications

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“…Most of the information that does exist on PAH metabolites has been obtained in the context of identifying transient metabolites formed by isolates during growth on the parent compound (13,33,35,40) or metabolites formed by mutants of wild-type degraders (4,40). Some bacteria, however, are capable of transforming one or more PAH despite an inability to grow on or mineralize the PAH in question (1,20,31,47).…”

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confidence: 99%

Products from the Incomplete Metabolism of Pyrene by Polycyclic Aromatic Hydrocarbon-Degrading Bacteria

Kazunga

Aitken

2000

Appl Environ Microbiol

161

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Pyrene is a regulated pollutant at sites contaminated with polycyclic aromatic hydrocarbons (PAH). It is mineralized by some bacteria but is also transformed to nonmineral products by a variety of other PAHdegrading bacteria. We examined the formation of such products by four bacterial strains and identified and further characterized the most apparently significant of these metabolites. Pseudomonas stutzeri strain P16 and Bacillus cereus strain P21 transformed pyrene primarily to cis-4,5-dihydro-4,5-dihydroxypyrene (PYRdHD), the first intermediate in the known pathway for aerobic bacterial mineralization of pyrene. Sphingomonas yanoikuyae strain R1 transformed pyrene to PYRdHD and pyrene-4,5-dione (PYRQ). Both strain R1 and Pseudomonas saccharophila strain P15 transform PYRdHD to PYRQ nearly stoichiometrically, suggesting that PYRQ is formed by oxidation of PYRdHD to 4,5-dihydroxypyrene and subsequent autoxidation of this metabolite. A pyrene-mineralizing organism, Mycobacterium strain PYR-1, also transforms PYRdHD to PYRQ at high initial concentrations of PYRdHD. However, strain PYR-1 is able to use both PYRdHD and PYRQ as growth substrates. PYRdHD strongly inhibited phenanthrene degradation by strains P15 and R1 but had only a minor effect on strains P16 and P21. At their aqueous saturation concentrations, both PYRdHD and PYRQ severely inhibited benzo[a]pyrene mineralization by strains P15 and R1. Collectively, these findings suggest that products derived from pyrene transformation have the potential to accumulate in PAH-contaminated systems and that such products can significantly influence the removal of other PAH. However, these products may be susceptible to subsequent degradation by organisms able to metabolize pyrene more extensively if such organisms are present in the system. Polycyclic aromatic hydrocarbons (PAH) are known to be degradable by a variety of soil bacteria (40). Consequently, the bioremediation of PAH contamination with naturally occurring microorganisms has been attempted at a number of sites (43,45). Most of the interest in the biodegradation of PAH in the field has been in the removal of the parent compounds, while most research on pure cultures of PAH-degrading bacteria has focused on their ability to grow on or mineralize specific PAH substrates. Relatively little attention has been paid to the potential formation of products from the partial transformation of PAH.Most of the information that does exist on PAH metabolites has been obtained in the context of identifying transient metabolites formed by isolates during growth on the parent compound (13,33,35,40) or metabolites formed by mutants of wild-type degraders (4, 40). Some bacteria, however, are capable of transforming one or more PAH despite an inability to grow on or mineralize the PAH in question (1,20,31,47). It is important to evaluate the products of such incomplete PAH metabolism because of their potential effects on PAH-degrading microorganisms or on potentially exposed human populations (44). Identification of common pr...

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confidence: 99%

Products from the Incomplete Metabolism of Pyrene by Polycyclic Aromatic Hydrocarbon-Degrading Bacteria

Kazunga

Aitken

2000

Appl Environ Microbiol

161

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“…This enzyme has not been isolated yet, but its selectivity was deduced from analysis of a mutant defective in dihydrodiol dehydrogenase catalyzing the second step in PAH degradation (14). Studies with this mutant strain showed that the S. yanoikuyae dioxygenase used a range of PAH substrates very similar to that found for PhnI, including the four-ring PAHs benz[a]anthracene and chrysene, as well as biphenyl, naphthalene, phenanthrene, and anthracene (6,14,15). Interestingly, both enzymes generate three cis-dihydrodiols from benz[a]anthracene, which were previously identified as derivatives bearing hydroxyl groups at the 1,2 positions, the 8,9 positions, and the 10,11 positions (14,15).…”

Section: Discussionmentioning

confidence: 99%

“…strain UW1 (43) and a Stenotrophomonas maltophilia strain (4). A mutant of Sphingomonas yanoikuyae (B8/36) was found to oxidize chrysene to (ϩ)-cis-3,4-dihydroxy-3,4-dihydrochrysene, a reaction most likely catalyzed by a ring-hydroxylating dioxygenase (6).…”

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Identification and Functional Analysis of Two Aromatic-Ring-Hydroxylating Dioxygenases from a Sphingomonas Strain That Degrades Various Polycyclic Aromatic Hydrocarbons

Demanèche

Meyer

Micoud

et al. 2004

Appl Environ Microbiol

137

119

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In this study, the enzymes involved in polycyclic aromatic hydrocarbon (PAH) degradation in the chrysenedegrading organism Sphingomonas sp. strain CHY-1 were investigated. [14 C]chrysene mineralization experiments showed that PAH-grown bacteria produced high levels of chrysene-catabolic activity. One PAH-induced protein displayed similarity with a ring-hydroxylating dioxygenase beta subunit, and a second PAH-induced protein displayed similarity with an extradiol dioxygenase. The genes encoding these proteins were cloned, and sequence analysis revealed two distinct loci containing clustered catabolic genes with strong similarities to corresponding genes found in Novosphingobium aromaticivorans F199. In the first locus, two genes potentially encoding a terminal dioxygenase component, designated PhnI, were followed by a gene coding for an aryl alcohol dehydrogenase (phnB). The second locus contained five genes encoding an extradiol dioxygenase (phnC), a ferredoxin (phnA3), another oxygenase component (PhnII), and an isomerase (phnD). PhnI was found to be capable of converting several PAHs, including chrysene, to the corresponding dihydrodiols. The activity of PhnI was greatly enhanced upon coexpression of genes encoding a ferredoxin (phnA3) and a reductase (phnA4). Disruption of the phnA1 a gene encoding the PhnI alpha subunit resulted in a mutant strain that had lost the ability to grow on PAHs. The recombinant PhnII enzyme overproduced in Escherichia coli functioned as a salicylate 1-hydroxylase. PhnII also used methylsalicylates and anthranilate as substrates. Our results indicated that a single enzyme (PhnI) was responsible for the initial attack of a range of PAHs, including chrysene, in strain CHY-1. Furthermore, the conversion of salicylate to catechol was catalyzed by a three-component oxygenase unrelated to known salicylate hydroxylases.

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“…This reaction was established with S. yanoikuyae B8/36, a mutant lacking biphenyl cis-dihydrodiol dehydrogenase [22]. Strain B8/36 oxidizes several aromatic substrates to cis-dihydrodiols including the polycyclic aromatic hydrocarbons naphthalene [16], anthracene [1,34], phenanthrene [34], chrysene [7], benz[a]anthracene [23,35] and benzo [a] pyrene [23]. The major cis-dihydrodiols formed from these substrates are shown in Fig.…”

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Purification, characterization, and crystallization of the components of a biphenyl dioxygenase system from Sphingobium yanoikuyae B1

Liu

Ferraro

et al. 2007

J Ind Microbiol Biotechnol

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Sphingobium yanoikuyae B1 initiates the catabolism of biphenyl by adding dioxygen to the aromatic nucleus to form (+)-cis-(2R, 3S)-dihydroxy-1-phenylcyclohexa-4,6-diene. The present study focuses on the biphenyl 2,3-dioxygenase system, which catalyzes the dioxygenation reaction. This enzyme has been shown to have a broad substrate range, catalyzing the dioxygenation of not only biphenyl, but also three- and four-ring polycyclic aromatic hydrocarbons. Extracts prepared from biphenyl-grown B1 cells contained three protein components that were required for the oxidation of biphenyl. The genes encoding the three components (bphA4, bphA3 and bphA1f,A2f) were expressed in Escherichia coli. Biotransformations of biphenyl, naphthalene, phenanthrene, and benzo[a]pyrene as substrates using the recombinant E. coli strain resulted in the formation of the expected cis-dihydrodiol products previously shown to be produced by biphenyl-induced strain B1. The three protein components were purified to apparent homogeneity and characterized in detail. The reductase component (bphA4), designated reductase(BPH-B1), was a 43 kD monomer containing one mol FAD/mol reductase(BPH-B1). The ferredoxin component (bphA3), designated ferredoxin(BPH-B1), was a 12 kD monomer containing approximately 2 g-atoms each of iron and acid-labile sulfur. The oxygenase component (bphA1f,A2f), designated oxygenase(BPH-B1), was a 217 kD heterotrimer consisting of alpha and beta subunits (approximately 51 and 21 kD, respectively). The iron and acid-labile sulfur contents of oxygenase(BPH-B1) per alphabeta were 2.4 and 1.8 g-atom per mol, respectively. Reduced ferredoxin(BPH-B1) and oxygenase(BPH-B1) each gave EPR signals typical of Rieske [2Fe-2S] proteins. Crystals of reductase(BPH-B1), ferredoxin(BPH-B1) and oxygenase(BPH-B1 )diffracted to 2.5 A, 2.0 A and 1.75 A, respectively. The structures of the three proteins are currently being determined.

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Bacterial dioxygenase-catalysed dihydroxylation and chemical resolution routes to enantiopure cis-dihydrodiols of chrysene

Cited by 28 publications

References 15 publications

Products from the Incomplete Metabolism of Pyrene by Polycyclic Aromatic Hydrocarbon-Degrading Bacteria

Products from the Incomplete Metabolism of Pyrene by Polycyclic Aromatic Hydrocarbon-Degrading Bacteria

Identification and Functional Analysis of Two Aromatic-Ring-Hydroxylating Dioxygenases from a Sphingomonas Strain That Degrades Various Polycyclic Aromatic Hydrocarbons

Purification, characterization, and crystallization of the components of a biphenyl dioxygenase system from Sphingobium yanoikuyae B1

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