Isolation and characterization of indene bioconversion genes from Rhodococcus strain I24

Treadway, Sheri L.; Yanagimachi, Kurt S.; Lankenau, E.; Lessard, Philip A.; Stephanopoulos, Gregory; Sinskey, Anthony J.

doi:10.1007/s002530051463

Cited by 71 publications

(59 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…A few polypeptides, including the two subunits of Pdo2, were synthesized only in the presence of a PAH, indicating that they are subjected to a regulation mechanism different from that controlling the enzymes induced by benzoate. Finally, we observed that some polypeptides were detectable only in pyrene-induced cells and that other proteins, including a dihydrodiol dehydrogenase, were found exclusively or predomi- The two dioxygenase terminal components described herein were found to be related to enzymes found in four actinomycete species, including two Rhodococcus strains (27,38), Nocardioides strain KP7 (34), and Mycobacterium strain PYR-1 (24). In previous reports, phylogenetic analysis indicated that these dioxygenases formed a cluster of enzymes distinct from other dioxygenases (24,34).…”

Section: Discussionmentioning

confidence: 79%

Identification of Pyrene-Induced Proteins in Mycobacterium sp. Strain 6PY1: Evidence for Two Ring-Hydroxylating Dioxygenases

Krivobok

Kuony²,

Meyer³

et al. 2003

J Bacteriol

191

151

View full text Add to dashboard Cite

In this study, the enzymes involved in polycyclic aromatic hydrocarbon (PAH) degradation were investigated in the pyrene-degrading Mycobacterium sp. strain 6PY1. [14 C]pyrene mineralization experiments showed that bacteria grown with either pyrene or phenanthrene produced high levels of pyrene-catabolic activity but that acetate-grown cells had no activity. As a means of identifying specific catabolic enzymes, protein extracts from bacteria grown on pyrene or on other carbon sources were analyzed by two-dimensional gel electrophoresis. Pyrene-induced proteins were tentatively identified by peptide sequence analysis. Half of them resembled enzymes known to be involved in phenanthrene degradation, with closest similarity to the corresponding enzymes from Nocardioides sp. strain KP7. The genes encoding the terminal components of two distinct ring-hydroxylating dioxygenases were cloned. Sequence analysis revealed that the two enzymes, designated Pdo1 and Pdo2, belong to a subfamily of dioxygenases found exclusively in gram-positive bacteria. When overproduced in Escherichia coli, Pdo1 and Pdo2 showed distinctive selectivities towards PAH substrates, with the former enzyme catalyzing the dihydroxylation of both pyrene and phenanthrene and the latter preferentially oxidizing phenanthrene. The catalytic activity of the Pdo2 enzyme was dramatically enhanced when electron carrier proteins of the phenanthrene dioxygenase from strain KP7 were coexpressed in recombinant cells. The Pdo2 enzyme was purified as a brown protein consisting of two types of subunits with M r s of about 52,000 and 20,000. Immunoblot analysis of cell extracts from strain 6PY1 revealed that Pdo1 was present in cells grown on benzoate, phenanthrene, or pyrene and absent in acetate-grown cells. In contrast, Pdo2 could be detected only in PAH-grown cells. These results indicated that the two enzymes were differentially regulated depending on the carbon source used for growth.Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants in soils and sediments and are of environmental concern because of their mutagenic and/or carcinogenic effects. While low-molecular-weight PAHs (composed of two or three rings) are readily degraded by bacteria, PAHs consisting of four rings or more are recalcitrant to biodegradation and persist in the environment (6, 41). The biodegradation of lowmolecular-weight PAHs, especially naphthalene, has been extensively studied with pseudomonads, leading to a good understanding of the bacterial catabolic pathway (42). On the other hand, relatively little information is available on the metabolism of high-molecular-weight PAHs (20). A number of bacterial isolates capable of pyrene mineralization have been described. Most of them are actinomycetes and belong to the genus Mycobacterium (2, 7, 37), Rhodococcus (4, 40), or Gordonia (21). A few pyrene-degrading strains have been identified as gram-negative species, including Stenotrophomonas maltophilia, Pseudomonas fluorescens (3), Sphingomonas paucimobilis (22), and Burkholder...

show abstract

Section: Discussionmentioning

confidence: 79%

Identification of Pyrene-Induced Proteins in Mycobacterium sp. Strain 6PY1: Evidence for Two Ring-Hydroxylating Dioxygenases

Krivobok

Kuony²,

Meyer³

et al. 2003

J Bacteriol

191

151

View full text Add to dashboard Cite

show abstract

“…Sequence alignments indicated that PhtB showed considerable divergence from A. keyseri 12B PhtB (Eaton, 2001), yet was related to Terrabacter sp. DBF63 PhtB (Habe et al, 2003) and other dihydrodiol dehydrogenases (Larkin et al, 1999;Masai et al, 1995;Saito et al, 2000;Treadway et al, 1999). Phylogenetic analysis of PhtB suggests, however, that the protein is closely related to PhtB of both Terrabacter sp.…”

Section: Discussionmentioning

confidence: 99%

Novel organization of genes in a phthalate degradation operon of Mycobacterium vanbaalenii PYR-1

et al. 2004

View full text Add to dashboard Cite

Mycobacterium vanbaalenii PYR-1 is capable of degrading polycyclic aromatic hydrocarbons (PAHs) to ring cleavage metabolites. This study identified and characterized a putative phthalate degradation operon in the M. vanbaalenii PYR-1 genome. A putative regulatory protein ( phtR) was encoded divergently with five tandem genes: phthalate dioxygenase large subunit ( phtAa), small subunit ( phtAb), phthalate dihydrodiol dehydrogenase ( phtB), phthalate dioxygenase ferredoxin subunit ( phtAc) and phthalate dioxygenase ferredoxin reductase ( phtAd ). A 6?7 kb EcoRI fragment containing these genes was cloned into Escherichia coli and converted phthalate to 3,4-dihydroxyphthalate. Homologues to the operon region were detected in a number of PAH-degrading Mycobacterium spp. isolated from various geographical locations. The operon differs from those of other Gram-positive bacteria in both the placement and orientation of the regulatory gene. In addition, the M. vanbaalenii PYR-1 pht operon contains no decarboxylase gene and none was identified within a 37 kb region containing the operon. This study is the first report of a phthalate degradation operon in Mycobacterium spp.

show abstract

“…Phylogenetic analysis of these oxygenase components, NidA, NidA3 and PhtAa, showed that they clustered with a new group of α subunits found in Nocardioides, Rhodococcus, Terrabacter, Arthrobacter and other Mycobacterium spp. [20][21][22][23][24][25]. Interestingly, RHO genes found in this group of bacteria appear to have features in common; genes for ETC components are not always closely positioned with oxygenase genes (genetic discreteness) and limited numbers of ferredoxin and reductase components are shared by multiple oxygenases (numerical imbalance).…”

Section: Introductionmentioning

confidence: 97%

A New Classification System for Bacterial Rieske Non-Heme Iron Aromatic Ring-Hydroxylating Oxygenases

Kweon¹,

Kim²,

Baek³

et al. 2011

Recent Advances in Biochemistry

View full text Add to dashboard Cite

Background: Rieske non-heme iron aromatic ring-hydroxylating oxygenases (RHOs) are multi-component enzyme systems that are remarkably diverse in bacteria isolated from diverse habitats. Since the first classification in 1990, there has been a need to devise a new classification scheme for these enzymes because many RHOs have been discovered, which do not belong to any group in the previous classification. Here, we present a scheme for classification of RHOs reflecting new sequence information and interactions between RHO enzyme components.

show abstract

Isolation and characterization of indene bioconversion genes from Rhodococcus strain I24

Cited by 71 publications

References 27 publications

Identification of Pyrene-Induced Proteins in Mycobacterium sp. Strain 6PY1: Evidence for Two Ring-Hydroxylating Dioxygenases

Identification of Pyrene-Induced Proteins in Mycobacterium sp. Strain 6PY1: Evidence for Two Ring-Hydroxylating Dioxygenases

Novel organization of genes in a phthalate degradation operon of Mycobacterium vanbaalenii PYR-1

A New Classification System for Bacterial Rieske Non-Heme Iron Aromatic Ring-Hydroxylating Oxygenases

Contact Info

Product

Resources

About