NADPH-dependent reductive ortho dehalogenation of 2,4-dichlorobenzoic acid in Corynebacterium sepedonicum KZ-4 and Coryneform bacterium strainNTB-1 via 2,4-dichlorobenzoyl coenzyme A

Journal of Bacteriology - Tập 178 Số 9 - Trang 2656-2661 - 1996
V. Romanov1, Robert P. Hausinger1
1Center for Microbial Ecology, Michigan State University, East Lansing 48824, USA

Tóm tắt

Corynebacterium sepedonicum KZ-4, described earlier as a strain capable of growth on 2,4-dichlorobenzoate (G.M. Zaitsev and Y.N. Karasevich, Mikrobiologiya 54:356-369, 1985), is known to metabolize this substrate via 4-hydroxybenzoate and protocatechuate, and evidence consistent with an initial reductive dechlorination step to form 4-chlorobenzoate was found in another coryneform bacterium, strain NTB-1 (W.J.J. van den Tweel, J.B. Kok, and J.A.M. de Bont, Appl. Environ. Microbiol. 53:810-815, 1987). 2-Chloro-4-fluorobenzoate was found to be converted stoichiometrically to 4-fluorobenzoate by resting cells of strain KZ-4, compatible with a reductive process. Experiments with cell extracts demonstrated that Mg - ATP and coenzyme A (CoA) were required to stimulate reductive dehalogenation, consistent with the intermediacy of 2-chloro-4-fluoro-benzoyl-CoA and 2,4-dichlorobenzoyl-CoA thioesters. 2,4-Dichlorobenzoyl-CoA was shown to be converted to 4-chlorobenzoyl-CoA in a novel NADPH-dependent reaction in extracts of both KZ-4 and NTB-1. In addition to the ligase and reductive dehalogenase activities, hydrolytic 4-chlorobenzoyl-CoA dehalogenase and thioesterase activities, 4-hydroxybenzoate 3-monooxygenase, and protocatechuate 3,4-dioxygenase activities were demonstrated to be present in the soluble fraction of KZ-4 extracts following ultracentrifugation. We propose that the pathway for 2,4-dichlorobenzoate catabolism in strains KZ-4 and NTB-1 involves formation of 2,4-dichlorobenzoyl-CoA, NADPH-dependent ortho dehalogenation yielding 4-chlorobenzoyl-CoA, hydrolytic removal of chlorine from the para position to generate 4-hydroxybenzoyl-CoA, hydrolysis to form 4-hydroxybenzoate, oxidation to yield protocatechuate, and oxidative ring cleavage.

Từ khóa


Tài liệu tham khảo

Bhat , G. B. , K. Iwase , B. C. Hummel , and P. G. Walfish . 1989 . Kinetic characteristics of a thioredoxin-activated rat hepatic and renal low-Km iodothyronine 5~-deiodinase . Biochem. J. 258 : 785 - 792 .

Bradford , M. 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 .

Chang , K. - H. , P.-H. Liang , W. Beck , J. D. Scholten , and D. Dunaway-Mariano . 1992 . Isolation and characterization of the three polypeptide components of 4-chlorobenzoate dehalogenase from Pseudomonas sp. strain CBS-3 . Biochemistry 31 : 5605 - 5610 .

Crooks , G. P. , and S. D. Copley . 1994 . Purification and characterization of 4-chlorobenzoyl CoA dehalogenase from Arthrobacter sp. strain 4-CB1 . Biochemistry 33 : 11645 - 11649 .

Dorn , E. , and H. J. Knackmuss . 1978 . Chemical structure and biodegradability of halogenated aromatic compounds. Two catechol 1,2-dioxygenases from a 3-chlorobenzoate-grown pseudomonad . Biochem. J. 174 : 73 - 84 .

Elder , D. J. E. , and D. J. Kelly . 1994 . The bacterial degradation of benzoic acid and benzenoid compounds under anaerobic conditions: unifying trends and new perspectives . FEMS Microbiol. Rev. 13 : 441 - 468 .

Entsch , B. , D. P. Ballou , and V. Massey . 1976 . Flavin-oxygen derivatives involved in hydroxylation by p-hydroxybenzoate hydroxylase . J. Biol. Chem. 251 : 2550 - 2563 .

Fetzner , S. , and F. Lingens . 1994 . Bacterial dehalogenases: biochemistry, genetics, and biotechnological applications . Microbiol. Rev. 58 : 641 - 685 .

Fujisawa , H. 1970 . Protocatechuate 3,4-dioxygenase . Methods Enzymol. 17A : 526 - 529 .

Furukawa , K. , and A. M. Chakrabarty . 1982 . Involvement of plasmids in total degradation of chlorinated biphenyls . Appl. Environ. Microbiol. 44 : 619 - 626 .

Goswami , A. , and I. N. Rosenberg . 1981 . Ferredoxin and ferredoxin reductase activities in bovine thyroid. Possible relationship to iodotyrosine deiodinase . J. Biol. Chem. 256 : 893 - 899 .

Groenewegen , P. E. J. , W. J. J. van den Tweel , and J. A. M. de Bont . 1992 . Anaerobic bioformation of 4-hydroxybenzoate from 4-chlorobenzoate by coryneform bacterium NTB-1 . Appl. Microbiol. Biotechnol. 36 : 541 - 547 .

Guirard B. M. and E. E. Snell. 1981. Biochemical factors in growth p. 79-111. In P. Gerhardt R. G. E. Murray R. N. Costilow E. W. Nester W. A. Wood N. R. Krieg and G. B. Phillips (ed.) Manual of methods for general bacteriology. American Society for Microbiology Washington D.C.

Habig , W. H. , M. J. Pabst , and W. B. Jacoby . 1974 . Glutathione S-transferases. The first enzymatic step in mercapturic acid formation . J. Biol. Chem. 249 : 7130 - 7139 .

Häggblom , M. M. 1992 . Microbial breakdown of halogenated aromatic pesticides and related compounds . FEMS Microbiol. Rev. 103 : 29 - 72 .

Hickey , W. J. , D. B. Searles , and D. D. Focht . 1993 . Enhanced mineralization of polychlorinated biphenyls in soil inoculated with chlorobenzoate-degrading bacteria . Appl. Environ. Microbiol. 59 : 1194 - 1200 .

Huang , Z. - H. , B.-J. Shyong , D. A. Gage , K. R. Noon , and J. Allison . 1994 . N-Alkylnicotinium halides: a class of cationic matrix additives for enhancing the sensitivity in negative ion fast-atom bombardment mass spectrometry of polyanionic analytes . J. Am. Soc. Mass Spectrom. 5 : 935 - 948 .

Janssen , D. B. , F. Pries , and J. R. van der Ploeg . 1994 . Genetics and biochemistry of dehalogenating enzymes. Annu . Rev. Microbiol. 48 : 163 - 191 .

Koch , J. , W. Eisenreich , A. Bacher , and G. Fuchs . 1993 . Products of enzymatic reduction of benzoyl-CoA, a key reaction in anaerobic aromatic metabolism . Eur. J. Biochem. 211 : 649 - 661 .

Li , S. , and L. P. Wackett . 1993 . Reductive dehalogenation by cytochrome P450CAM: substrate binding and catalysis . Biochemistry 32 : 9355 - 9361 .

Liu , R. - Q. , P.-H. Liang , J. Scholten , and D. Dunaway-Mariano . 1995 . Transient state kinetic analysis of the chemical intermediates formed in the enzymatic dehalogenation of 4-chlorobenzoyl coenzyme A . J. Am. Chem. Soc. 117 : 5003 - 5004 .

Lochmeyer , C. , J. Koch , and G. Fuchs . 1992 . Anaerobic degradation of 2-aminobenzoic acid (anthranilic acid) via benzoyl-coenzyme A (CoA) and cyclohex-1-enecarboxyl-CoA in a denitrifying bacterium . J. Bacteriol. 174 : 3621 - 3628 .

Löffler , F. , F. Lingens , and R. Müller . 1995 . Dehalogenation of 4-chlorobenzoate. Characterisation of 4-chlorobenzoyl-coenzyme A dehalogenase from Pseudomonas sp. CBS3 . Biodegradation 6 : 203 - 212 .

Löffler , F. , R. Müller , and F. Lingens . 1991 . Dehalogenation of 4-chlorobenzoate by 4-chlorobenzoate dehalogenase from Pseudomonas sp. CBS3: an ATP/coenzyme A dependent reaction . Biochem. Biophys. Res. Commun. 176 : 1106 - 1111 .

Löffler F. E. and J. M. Tiedje. 1995. Characterization of components involved in respiratory dechlorination of 3-chlorobenzoate abstr. Q-98 p. 416. In Abstracts of the 95th General Meeting of the American Society for Microbiology 1995. American Society for Microbiology Washington D.C.

Merkel , S. M. , A. E. Eberhard , J. Gibson , and C. S. Harwood . 1989 . Involvement of coenzyme A thioesters in anaerobic metabolism of 4-hydroxybenzoate by Rhodopseudomonas palustris . J. Bacteriol. 171 : 1 - 7 .

Mohn , W. W. , and J. M. Tiedje . 1992 . Microbial reductive dehalogenation . Microbiol. Rev. 56 : 482 - 507 .

Ni , S. , J. K. Frederickson , and L. Xun . 1995 . Purification and characterization of a novel 3-chlorobenzoate reductive dehalogenase from the cytoplasmic membrane of Desulfomonile tiedjei DCB-1 . J. Bacteriol. 177 : 5135 - 5139 .

Nozaki , M. 1970 . Metapyrocatechase (Pseudomonas) . Methods Enzymol. 17A : 522 - 525 .

Nozawa , T. , and Y. Maruyama . 1988 . Anaerobic metabolism of phthalate and other aromatic compounds by a denitrifying bacterium . J. Bacteriol. 170 : 5778 - 5784 .

Orser , C. S. , J. Dutton , C. Lange , P. Jablonski , L. Xun , and M. Hargis . 1993 . Characterization of a Flavobacterium glutathione S-transferase gene involved in reductive dechlorination . J. Bacteriol. 175 : 2640 - 2644 .

Romanov , V. , and R. P. Hausinger . 1994 . Pseudomonas aeruginosa 142 uses a three-component ortho-halobenzoate 1,2-dioxygenase for metabolism of 2,4-dichloro- and 2-chlorobenzoate . J. Bacteriol. 176 : 3368 - 3374 .

Romanov V. and R. P. Hausinger. 1995. Reductive dehalogenation of 2 4- dichlorobenzoate by Corynebacterium sepedonicum KZ-4 abstr. Q-99 p. 417. In Abstracts of the 95th General Meeting of the American Society for Microbiology 1995. American Society for Microbiology Washington D.C.

Schmitz , A. , K. -H. Garteman , J. Fiedler , E. Grund , and R. Eichenlaub . 1992 . Cloning, and sequence analysis of genes for dehalogenation of 4-chlorobenzoate from Arthrobacter sp. strain SU . Appl. Environ. Microbiol. 58 : 4068 - 4071 .

Scholten , J. D. , K. -H. Chang , P. C. Babbitt , H. Charest , M. Sylvestre , and D. Dunaway-Mariano . 1991 . Novel enzymic hydrolytic dehalogenation of a chlorinated aromatic . Science 253 : 182 - 185 .

Shimao , M. , S. Onishi , S. Mizumori , N. Kato , and C. Sakazawa . 1989 . Degradation of 4-chlorobenzoate by facultatively alkalophilic Arthrobacter sp. strain SB-8 . Appl. Environ. Microbiol. 55 : 478 - 482 .

Thiele , J. , R. Müller , and F. Lingens . 1987 . Initial characterization of 4-chlorobenzoate dehalogenase from Pseudomonas sp. CBS3 . FEMS Microbiol. Lett. 41 : 115 - 119 .

Tsoi T. and J. M. Tiedje. 1992. Personal communication.

van den Tweel , W. J. J. , J. B. Kok , and J. A. M. de Bont . 1987 . Reductive dechlorination of 2,4-dichlorobenzoate to 4-chlorobenzoate and hydrolytic dehalogenation of 4-chloro-, 4-bromo-, and 4-iodobenzoate by Alcaligenes denitrificans NTB-1 . Appl. Environ. Microbiol. 53 : 810 - 815 .

Xun , L. , E. Topp , and C. S. Orser . 1992 . Purification and characterization of a tetrachloro-p-hydroquinone reductive dehalogenase from a Flavobacterium sp . J. Bacteriol. 174 : 8003 - 8007 .

Yang , G. , P. -H. Liang , and D. Dunaway-Mariano . 1994 . Evidence for nucleophilic catalysis in the aromatic substitution reaction catalyzed by (4- chlorobenzoyl) coenzyme A dehalogenase . Biochemistry 33 : 8527 - 8531 .

Zaitsev , G. M. , and Y. N. Karasevich . 1985 . Preparatory metabolism of 4-chlorobenzoic and 2,4-dichlorobenzoic acids in Corynebacterium sepedonicum . Mikrobiologiya 54 : 356 - 359 . (In Russian.)