RESEARCH ARTICLE

A newly defined dioxygenase system from Mycobacterium vanbaalenii PYR-1 endowed with an enhanced activity of dihydroxylation of high-molecular-weight polyaromatic hydrocarbons

  • Yiquan Wu 1,2 ,
  • Ying Xu , 1,2 ,
  • Ningyi Zhou 1,2
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  • 1. State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China
  • 2. Joint International Research Laboratory of Metabolic & Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, China

Received date: 20 Aug 2019

Revised date: 18 Sep 2019

Accepted date: 22 Sep 2019

Published date: 15 Feb 2020

Copyright

2019 Higher Education Press and Springer-Verlag GmbH Germany, part of Springer Nature

Highlights

Mycobacterium vanbaalenii PYR-1 utilizes PAHs at different rates.

• Both NidA3B3 and FNidA3B3 catalyze high-molecular-weight PAHs dihydroxylation.

• NidA3B3 shows an enhanced activity with an endogenous electron transport chain.

• FNidA3B3 has an enhanced activity by 50% approximately compared with NidA3B3.

• FNidA3B3/NidA3B3 and NidAB in the same strain come from different ancestors.

Abstract

NidA3B3 is a terminal dioxygenase whose favorable substrates are high-molecular-weight polyaromatic hydrocarbons (PAHs) from Mycobacterium vanbaalenii PYR-1, a powerful PAHs degradation strain. NidA3B3 was reported to incorporate a dioxygen into the benzene ring of PAHs when equipped with an exogenous electron transport chain components PhdCD from Nocardioides sp. strain KP7 by biotransformation, but this enzyme system was not particularly efficient. In this study, strain PYR-1 was confirmed to utilize four different PAHs at different growth rates. When PhtAcAd, an endogenous electron transport chain of a phthalate dioxygenase system, was substituted for PhdCD to couple with NidA3B3, the specific activity to convert phenanthrene by strain BL21(DE3) [pNidA3B3-PhAcAd] was 0.15±0.03 U/mg, but the specific activity of strain BL21(DE3) [pNidA3B3-PhdCD] was only 0.025±0.006 U/mg. In addition, FNidA3, encoded by a newly defined ORF, has a prolonged 19-amino acid sequence at the N-terminal compared with NidA3. FNidA3B3 increased the activity by 50% approximately than NidA3B3 when using PhtAcAd. Components of the electron transport chain PhtAc and PhtAd were purified and characterized. The Km, kcat, kcat/Km values of the PhtAd were 123±26.9 M, 503±49.9 min1, 4.1 M1·min1, respectively. And the Km, kcat, kcat/Km values of the ferredoxin PhtAc were 52.5±9.7 M, 3.8±0.19 min1 and 0.07 M1·min1, respectively. Basing on the phylogenetic analysis, NidA3/FNidA3 were far from its isoenzyme NidA from the same strain. Combining their primary differences of transcriptional pattern in vivo, it indicated that the functionally similar Rieske dioxygenases NidA3B3/FNidA3B3 and NidAB might originate from different ancestors.

Cite this article

Yiquan Wu , Ying Xu , Ningyi Zhou . A newly defined dioxygenase system from Mycobacterium vanbaalenii PYR-1 endowed with an enhanced activity of dihydroxylation of high-molecular-weight polyaromatic hydrocarbons[J]. Frontiers of Environmental Science & Engineering, 2020 , 14(1) : 14 . DOI: 10.1007/s11783-019-1193-5

Acknowledgements

This work is supported by the National Key R&D Program of China (Grant No. 2018YFC0309800), National Natural Science Foundation of China (Grant No. 31570100) and Shanghai Science and Technology Commission Scientific Research Project (No. 17JC1403300).
1
Barry S M, Challis G L (2013). Mechanism and catalytic diversity of Rieske non-heme iron-dependent oxygenases. ACS Catalysis, 3(10): 2362–2370

DOI PMID

2
Boldrin B, Tiehm A, Fritzsche C (1993). Degradation of phenanthrene, fluorene, fluoranthene, and pyrene by a Mycobacterium sp. Applied and Environmental Microbiology, 59(6): 1927–1930

PMID

3
Boström C E, Gerde P, Hanberg A, Jernström B, Johansson C, Kyrklund T, Rannug A, Törnqvist M, Victorin K, Westerholm R (2002). Cancer risk assessment, indicators, and guidelines for polycyclic aromatic hydrocarbons in the ambient air. Environmental Health Perspectives, 110(Suppl 3): 451–488

PMID

4
Capyk J K, D’Angelo I, Strynadka N C, Eltis L D (2009). Characterization of 3-ketosteroid 9-hydroxylase, a Rieske oxygenase in the cholesterol degradation pathway of Mycobacterium tuberculosis. Journal of biological chemistry, 284(15): 9937–9946

DOI PMID

5
Cébron A, Norini M P, Beguiristain T, Leyval C (2008). Real-Time PCR quantification of PAH-ring hydroxylating dioxygenase (PAH-RHDalpha) genes from Gram positive and Gram negative bacteria in soil and sediment samples. Journal of Microbiological Methods, 73(2): 148–159

DOI PMID

6
Dean-Ross D, Cerniglia C E (1996). Degradation of pyrene by Mycobacterium flavescens. Applied Microbiology and Biotechnology, 46(3): 307–312

DOI PMID

7
DeBruyn J M, Mead T J, Sayler G S (2012). Horizontal transfer of PAH catabolism genes in Mycobacterium: evidence from comparative genomics and isolated pyrene-degrading bacteria. Environmental Science & Technology, 46(1): 99–106

DOI PMID

8
Ensley B D, Gibson D T (1983). Naphthalene dioxygenase: Purification and properties of a terminal oxygenase component. Journal of Bacteriology, 155(2): 505–511

PMID

9
Fischer F, Raimondi D, Aliverti A, Zanetti G (2002). Mycobacterium tuberculosis FprA, a novel bacterial NADPH-ferredoxin reductase. European Journal of Biochemistry, 269(12): 3005–3013

DOI PMID

10
Heitkamp M A, Franklin W, Cerniglia C E (1988). Microbial metabolism of polycyclic aromatic hydrocarbons: Isolation and characterization of a pyrene-degrading bacterium. Applied and Environmental Microbiology, 54(10): 2549–2555

PMID

11
Karlsson A, Parales J V, Parales R E, Gibson D T, Eklund H, Ramaswamy S (2003). Crystal structure of naphthalene dioxygenase: Side-on binding of dioxygen to iron. Science, 299(5609): 1039–1042

DOI PMID

12
Khan A A, Wang R F, Cao W W, Doerge D R, Wennerstrom D, Cerniglia C E (2001). Molecular cloning, nucleotide sequence, and expression of genes encoding a polycyclic aromatic ring dioxygenase from Mycobacterium sp. strain PYR-1. Applied and Environmental Microbiology, 67(8): 3577–3585

DOI PMID

13
Kim S J, Kweon O, Freeman J P, Jones R C, Adjei M D, Jhoo J W, Edmondson R D, Cerniglia C E (2006). Molecular cloning and expression of genes encoding a novel dioxygenase involved in low- and high-molecular-weight polycyclic aromatic hydrocarbon degradation in Mycobacterium vanbaalenii PYR-1. Applied and Environmental Microbiology, 72(2): 1045–1054

DOI PMID

14
Kim S J, Kweon O, Jones R C, Edmondson R D, Cerniglia C E (2008). Genomic analysis of polycyclic aromatic hydrocarbon degradation in Mycobacterium vanbaalenii PYR-1. Biodegradation, 19(6): 859–881

DOI PMID

15
Kim S J, Kweon O, Jones R C, Freeman J P, Edmondson R D, Cerniglia C E (2007). Complete and integrated pyrene degradation pathway in Mycobacterium vanbaalenii PYR-1 based on systems biology. Journal of Bacteriology, 189(2): 464–472

DOI PMID

16
Kweon O, Kim S J, Freeman J P, Song J, Baek S, Cerniglia C E (2010). Substrate specificity and structural characteristics of the novel Rieske nonheme iron aromatic ring-hydroxylating oxygenases NidAB and NidA3B3 from Mycobacterium vanbaalenii PYR-1. mBio, 1(2): e00135-10

DOI PMID

17
Kweon O, Kim S J, Holland R D, Chen H, Kim D W, Gao Y, Yu L R, Baek S, Baek D H, Ahn H, Cerniglia C E (2011). Polycyclic aromatic hydrocarbon metabolic network in Mycobacterium vanbaalenii PYR-1. Journal of Bacteriology, 193(17): 4326–4337

DOI PMID

18
Kweon O, Kim S J, Jones R C, Freeman J P, Adjei M D, Edmondson R D, Cerniglia C E (2007). A polyomic approach to elucidate the fluoranthene-degradative pathway in Mycobacterium vanbaalenii PYR-1. Journal of Bacteriology, 189(13): 4635–4647

DOI PMID

19
Kweon O, Kim S J, Kim D W, Kim J M, Kim H L, Ahn Y, Sutherland J B, Cerniglia C E (2014). Pleiotropic and epistatic behavior of a ring-hydroxylating oxygenase system in the polycyclic aromatic hydrocarbon metabolic network from Mycobacterium vanbaalenii PYR-1. Journal of Bacteriology, 196(19): 3503–3515

DOI PMID

20
Lama A, Pawaria S, Bidon-Chanal A, Anand A, Gelpí J L, Arya S, Martí M, Estrin D A, Luque F J, Dikshit K L (2009). Role of Pre-A motif in nitric oxide scavenging by truncated hemoglobin, HbN, of Mycobacterium tuberculosis. Journal of biological chemistry, 284(21): 14457–14468

DOI PMID

21
McLean K, Dunford A, Sabri M, Neeli R, Girvan H, Balding P, Leys D, Seward H, Marshall K, Munro A (2006). CYP121, CYP51 and Associated Redox Systems inMycobacterium tuberculosis: Towards Deconvoluting Enzymology of P450 Systems in a Human Pathogen. London: Portland Press Limited

22
Moody J D, Freeman J P, Doerge D R, Cerniglia C E (2001). Degradation of phenanthrene and anthracene by cell suspensions of Mycobacterium sp. strain PYR-1. Applied and Environmental Microbiology, 67(4): 1476–1483

DOI PMID

23
Moody J D, Freeman J P, Fu P P, Cerniglia C E (2004). Degradation of benzo[a]pyrene by Mycobacterium vanbaalenii PYR-1. Applied and Environmental Microbiology, 70(1): 340–345

DOI PMID

24
Saito A, Iwabuchi T, Harayama S (2000). A novel phenanthrene dioxygenase from Nocardioides sp. Strain KP7: Expression in Escherichia coli. Journal of Bacteriology, 182(8): 2134–2141

DOI PMID

25
Schneider J, Grosser R, Jayasimhulu K, Xue W, Warshawsky D (1996). Degradation of pyrene, benz[a]anthracene, and benzo[a]pyrene by Mycobacterium sp. strain RJGII-135, isolated from a former coal gasification site. Applied and Environmental Microbiology, 62(1): 13–19

PMID

26
Siebold C, Flükiger K, Beutler R, Erni B (2001). Carbohydrate transporters of the bacterial phosphoenolpyruvate: sugar phosphotransferase system (PTS). FEBS Letters, 504(3): 104–111

DOI PMID

27
Stingley R L, Brezna B, Khan A A, Cerniglia C E (2004). Novel organization of genes in a phthalate degradation operon of Mycobacterium vanbaalenii PYR-1. Microbiology, 150(11): 3749–3761

DOI PMID

28
Stone E A, Lough G C, Schauer J J, Praveen P, Corrigan C, Ramanathan V (2007). Understanding the origin of black carbon in the atmospheric brown cloud over the Indian Ocean. Journal of Geophysical Research: Atmospheres, 112(D22): D22S23, 1-10

DOI

29
Wang L, Gao Y Z, Zhao H, Xu Y, Zhou N Y (2019). Biodegradation of 2-bromonitrobenzene by Pseudomonas stutzeri ZWLR2–1. International Biodeterioration & Biodegradation, 138: 87–91

DOI

30
Wolfe M D, Parales J V, Gibson D T, Lipscomb J D (2001). Single turnover chemistry and regulation of O2 activation by the oxygenase component of naphthalene 1,2-dioxygenase. Journal of biological chemistry, 276(3): 1945–1953

DOI PMID

31
Xu Y, Zhou N Y (2017). Microbial remediation of aromatics-contaminated soil. Frontiers of Environmental Science & Engineering, 11(2): 1–9

DOI

32
Yu C L, Liu W, Ferraro D J, Brown E N, Parales J V, Ramaswamy S, Zylstra G J, Gibson D T, Parales R E (2007). Purification, characterization, and crystallization of the components of a biphenyl dioxygenase system from Sphingobium yanoikuyae B1. Journal of Industrial Microbiology & Biotechnology, 34(4): 311–324

DOI PMID

33
Zhang Y, Tao S, Shen H, Ma J (2009). Inhalation exposure to ambient polycyclic aromatic hydrocarbons and lung cancer risk of Chinese population. Proceedings of the National Academy of Sciences of the United States of America, 106(50): 21063–21067

DOI PMID

34
Zhou H W, Guo C L, Wong Y S, Tam N F Y (2006). Genetic diversity of dioxygenase genes in polycyclic aromatic hydrocarbon-degrading bacteria isolated from mangrove sediments. FEMS Microbiology Letters, 262(2): 148–157

DOI PMID

35
Zhou N Y, Al-Dulayymi J, Baird M S, Williams P A (2002). Salicylate 5-hydroxylase from Ralstonia sp. strain U2: A monooxygenase with close relationships to and shared electron transport proteins with naphthalene dioxygenase. Journal of Bacteriology, 184(6): 1547–1555

DOI PMID

36
Zink G, Lorber K E (1995). Mass spectral identification of metabolites formed by microbial degradation of polycyclic aromatic hydrocarbons (PAH). Chemosphere, 31(9): 4077–4084

DOI PMID

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