Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate
A gene cluster containing a gene for maleylacetate reductase (EC 1.3.1.32) was cloned from Ralstonia eutropha 335T (DSM 531T), which is able to utilize 4-fluorobenzoate as sole carbon source. Sequencing of this gene cluster showed that the R. eutropha 335T maleylacetate reductase gene, macA, is part of a novel gene cluster, which is not related to the known maleylacetate-reductase-encoding gene clusters. It otherwise comprises a gene for a hypothetical membrane transport protein, macB, possibly co-transcribed with macA, and a presumed regulatory gene, macR, which is divergently transcribed from macBA. MacA was found to be most closely related to TftE, the maleylacetate reductase from Burkholderia cepacia AC1100 (62 % identical positions) and to a presumed maleylacetate reductase from a dinitrotoluene catabolic gene cluster from B. cepacia R34 (61 % identical positions). By expressing macA in Escherichia coli, it was confirmed that macA encodes a functional maleylacetate reductase. Purification of maleylacetate reductase from 4-fluorobenzoate-grown R. eutropha 335T cells allowed determination of the N-terminal sequence of the purified protein, which was shown to be identical to that predicted from the cloned macA gene, thus proving that the gene is, in fact, recruited for growth of R. eutropha 335T with this substrate.
AltschulS. F.,
MaddenT. L.,
SchafferA. A.,
ZhangJ.,
ZhangZ.,
MillerW.,
LipmanD. J.
1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402[CrossRef]
ArmengaudJ.,
TimmisK. N.,
WittichR.-M.
1999; A functional 4-hydroxysalicylate/hydroxyquinol degradative pathway gene cluster is linked to the initial dibenzo- p -dioxin pathway genes in Sphingomonas sp. strain RW1. J Bacteriol 181:3452–3461
BaitschD.,
SanduC.,
BrandischR.,
IgloiG. L.
2001; Gene cluster on pAO1 of Arthrobacter nicotinovorans involved in degradation of the plant alkaloid nicotine: cloning, purification, and characterization of 2,6-dihydroxypyridine 3-hydroxylase. J Bacteriol 183:5262–5267[CrossRef]
BradfordM. M.
1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254[CrossRef]
ChapmanP. J.,
RibbonsD. W.
1976; Metabolism of resorcinylic compounds by bacteria: alternative pathways for resorcinol catabolism in Pseudomonas putida
. J Bacteriol 125:985–998
DaubarasD. L.,
HershbergerC. D.,
KitanoK.,
ChakrabartyA. M.
1995; Sequence analysis of a gene cluster involved in metabolism of 2,4,5-trichlorophenoxyacetic acid by Burkholderia cepacia AC1100. Appl Environ Microbiol 61:1279–1289
DaubarasD. L.,
SaidoK.,
ChakrabartyA. M.
1996; Purification of hydroxyquinol 1,2-dioxygenase and maleylacetate reductase: the lower pathway of 2,4,5-trichlorophenoxyacetic acid metabolism by Burkholderia cepacia AC1100. Appl Environ Microbiol 62:4276–4279
DonR. H.,
PembertonJ. M.
1981; Properties of six pesticide degradation plasmids isolated from Alcaligenes paradoxus and Alcaligenes eutrophus
. J Bacteriol 145:681–686
DonR. H.,
WeightmanA. J.,
KnackmussH.-J.,
TimmisK. N.
1985; Transposon mutagenesis and cloning analysis of the pathways for degradation of 2,4-dichlorophenoxyacetate acid and 3-chlorobenzoate in Alcaligenes eutrophus JMP134 (pJP4). J Bacteriol 161:85–90
DuxburyJ. M.,
TiedjeJ. M.,
AlexanderM.,
DawsonJ. E.
1970; 2,4-D metabolism: enzymatic conversion of chloromaleylacetic acid to succinic acid. J Agric Food Chem 18:199–201[CrossRef]
EulbergD.,
KourbatovaE. M.,
GolovlevaL. A., SchlömannM. 1998; Evolutionary relationship between chlorocatechol catabolic enzymes from Rhodococcus opacus 1CP and their counterparts in Proteobacteria: sequence divergence and functional convergence. J Bacteriol 180:1082–1094
FeigelB. J.,
KnackmussH.-J.
1993; Syntrophic interactions during degradation of 4-aminobenzenesulfonic acid by a two species bacterial culture. Arch Microbiol 159:124–130[CrossRef]
FrantzB.,
ChakrabartyA. M.
1987; Organization and nucleotide sequence determination of a gene cluster involved in 3-chlorocatechol degradation. Proc Natl Acad Sci U S A 84:4460–4464[CrossRef]
FritscheK.
1998Molekularbiologische Untersuchungen zum Chlorphenolabbau durch Stamm S1, ein Proteobakterium der α-2-Untergruppe PhD thesis Martin-Luther-University Halle; Halle/Saale, Germany:
HenikoffS.,
HaughnG. W.,
CalvoJ. M.,
WallaceJ. C.
1988; A large family of bacterial activator proteins. Proc Natl Acad Sci U S A 85:6602–6606[CrossRef]
HinnerI.-S.
1998Biochemische und molekularbiologische Untersuchungen zu Lacton-Hydrolasen des bakteriellen Aromaten- und Halogenaromaten-Abbaus PhD thesis University of Stuttgart; Stuttgart, Germany:
JainR. K.,
DreisbachJ. H.,
SpainJ. C.
1994; Biodegradation of p -nitrophenol via 1,2,4-benzenetriol by an Arthrobacter sp. Appl Environ Microbiol 60:3030–3032
JonesK. H.,
TrudgillP. W.,
HopperD. J.
1995; Evidence of two pathways for the metabolism of phenol by Aspergillus fumigatus
. Arch Microbiol 163:176–181[CrossRef]
KanekoT.,
NakamuraY.,
SatoS.14 other authors2002; Complete genomic sequence of nitrogen-fixing symbiotic bacterium Bradyrhizobium japonicum USDA110. DNA Res 9:189–197[CrossRef]
KarasevichYu. N.,
IvoilovV. S.
1977; Preparatory metabolism of para -hydroxybenzoic acid in the yeast Candida tropicalis
. Microbiology (English translation of Mikrobiologiya) 46:687–695
KasbergT.,
DaubarasD. L.,
ChakrabartyA. M.,
ReinekeW.
1995; Evidence that operons tcb , tfd , and clc encode maleylacetate reductase, the fourth enzyme of the modified ortho pathway. J Bacteriol 177:3885–3889
KasbergT.,
SeibertV.,
ReinekeW, SchlömannM.1997; Cloning, characterization, and sequence analysis of the clcE gene encoding the maleylacetate reductase of Pseudomonas sp strain B13. J Bacteriol 179:3801–3803
LaemmliC. M.,
LeveauJ. H.,
ZehnderA. J., van der MeerJ. R. 2000; Characterization of a second tfd gene cluster for chlorophenol and chlorocatechol metabolism on plasmid pJP4 in Ralstonia eutropha
. J Bacteriol 182:4165–4172[CrossRef]
LatusM.,
SeitzH.-J.,
LingensF, EberspächerJ.1995; Purification and characterization of hydroxyquinol 1,2-dioxygenase from Azotobacter sp strain GP1. Appl Environ Microbiol 61:2453–2460
LiuS.,
OgawaN.,
MiyashitaK.
2001; The chlorocatechol degradative genes, tfdT-CDEF , of Burkholderia sp. strain NK8 are involved in chlorobenzoate degradation and induced by chlorobenzoates and chlorocatechols. Gene 268:207–214[CrossRef]
MiyauchiK.,
AdachiY.,
NagataY.,
TakagiM.
1999; Cloning and sequencing of a novel meta -cleavage dioxygenase gene whose product is involved in degradation of γ -hexachlorocyclohexane in Sphingomonas paucimobilis
. J Bacteriol 181:6712–6719
MoiseevaO. V.,
SolyanikovaI. P.,
KaschabekS. R.,
ThielM.,
GolovlevaL. A, GröningJ, SchlömannM. 2002; A new modified ortho cleavage pathway of 3-chlorocatechol degradation by Rhodococcus opacus 1CP: genetic and biochemical evidence. J Bacteriol 184:5282–5292[CrossRef]
PerkinsE. J.,
GordonM. P.,
CaceresO.,
LurquinP. F.
1990; Organization and sequence analysis of the 2,4-dichlorophenol hydroxylase and dichlorocatechol oxidative operons of plasmid pJP4. J Bacteriol 172:2351–2359
PlumeierI.,
HeimS.,
PieperD. H, Pérez-PantojaD.,
GonzálezB.2002; Importance of different tfd genes for degradation of chloroaromatics by Ralstonia eutropha JMP134. J Bacteriol 184:4054–4064[CrossRef]
SchlömannM. 1994; Evolution of chlorocatechol catabolic pathways. Conclusions to be drawn from comparisons of lactone hydrolases. Biodegradation 5:301–321[CrossRef]
SchlömannM.,
SchmidtE.,
KnackmussH.-J.
1990a; Different types of dienelactone hydrolase in 4-fluorobenzoate-utilizing bacteria. J Bacteriol 172:5112–5118
SchlömannM.,
FischerP.,
SchmidtE.,
KnackmussH.-J.
1990b; Enzymatic formation, stability, and spontaneous reactions of 4-fluoromuconolactone, a metabolite of the bacterial degradation of 4-fluorobenzoate. J Bacteriol 172:5119–5129
SeibertV., SchlömannM. 1996Cloning, sequencing and purification of maleylacetate reductase from Alcaligenes eutrophus 335.In Biospektrum (special issue) p 123 abstract PE143.;
SeibertV.,
KourbatovaE. M.,
GolovlevaL. M., SchlömannM. 1998; Characterization of the maleylacetate reductase MacA of Rhodococcus opacus 1CP and evidence for the presence of an isofunctional enzyme. J Bacteriol 180:3503–3508
StudierF. W.,
RosenbergA. H.,
DunnJ. J.,
DubendorffJ. W.
1990; Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol 185:60–89
ThompsonJ. D.,
GibsonT. J.,
PlewniakF.,
JeanmouginF.,
HigginsD. G.
1997; The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882[CrossRef]
van der MeerJ. R.,
EggenR. I. L.,
ZehnderA. J. B., de VosW. M. 1991; Sequence analysis of the Pseudomonas sp. strain P51 tcb gene cluster, which encodes metabolism of chlorinated catechols: evidence for specialization of catechol 1,2-dioxygenases for chlorinated substrates. J Bacteriol 173:2425–2434
van der MeerJ. R.,
de VosW. M.,
HarayamaS.,
ZehnderA. J. B.
1992; Molecular mechanisms of genetic adaptation to xenobiotic compounds. Microbiol Rev 56:677–694
WangY. Z.,
ZhouY.,
ZylstraG. J.
1995; Molecular analysis of isophthalate and terephthalate degradation by Comamonas testosteroni YZW-D. Environ Health Perspect 103:9–12[CrossRef]
WoodD. W.,
SetubalJ. C.,
KaulR.48 other authors2001; The genome of the natural genetic engineer Agrobacterium tumefaciens C58. Science 294:2317–2323[CrossRef]
ZaborinaO.,
LatusM.,
GolovlevaL. A.,
LingensF, EberspächerJ.1995; Purification and characterization of 6-chlorohydroxyquinol 1,2-dioxygenase from Streptomyces rochei 303: comparison with an analogous enzyme from Azotobacter sp. strain GP1. J Bacteriol 177:229–234
Characterization of a gene cluster encoding the maleylacetate reductase from Ralstonia eutropha 335T, an enzyme recruited for growth with 4-fluorobenzoate