Growth of bacteria on 3-nitropropionic acid as a sole source of carbon, nitrogen, and energy.
3-Nitropropionic acid (3NPA) is a widespread nitroaliphatic toxin found in a variety of legumes and fungi. Several enzymes have been reported that can transform the compound, but none led to the mineralization of 3NPA. We report here the isolation of bacteria that grow on 3NPA and its anion, propionate-3-nitronate (P3N), as the sole source of carbon, nitrogen, and energy. Experiments with resting cells, cell extracts, and purified enzymes indicate that the pathway involves conversion of 3NPA to P3N, which upon denitration yields malonic semialdehyde, nitrate, nitrite, and traces of H(2)O(2). Malonic semialdehyde is decarboxylated to acetyl coenzyme A. The gene that encodes the enzyme responsible for the denitration of P3N was cloned and expressed, and the enzyme was purified. Stoichiometry of the reaction indicates that the enzyme is a monooxygenase. The gene sequence is related to a large group of genes annotated as 2-nitropropane dioxygenases, but the P3N monooxygenase and closely related enzymes form a cluster within COG2070 that differs from previously characterized 2-nitropropane dioxygenases by their substrate specificities and reaction products. The results suggest that the P3N monooxygenases enable bacteria to exploit 3NPA in natural habitats as a growth substrate.
Nishino
Shirley F
SF
School of Civil and Environmental Engineering, Georgia Institute of Technology, 311 Ferst Drive, Atlanta, GA 30332-0512, USA.
Shin
Kwanghee A
KA
Payne
Rayford B
RB
Spain
Jim C
JC
D001419:Bacteria / Q000302:isolation & purification / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics; D002244:Carbon / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004734:Energy Metabolism / Q000235:genetics*; D015870:Gene Expression; D053858:Metabolic Networks and Pathways / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000235:genetics; D008969:Molecular Sequence Data; D009574:Nitro Compounds / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D010802:Phylogeny; D011422:Propionates / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D009574:Nitro Compounds; D011422:Propionates; D002244:Carbon; D006899:Mixed Function Oxygenases; D009584:Nitrogen; C015392:3-nitropropionic acid
Appl Environ Microbiol. 1998 Mar;64(3):1029-33
9501443
Nature. 2004 Jul 1;430(6995):98-101
15229604
Appl Environ Microbiol. 1993 Sep;59(9):3056-61
8215375
J Biol Chem. 2006 Jul 7;281(27):18660-7
16682407
J Bacteriol. 2002 May;184(9):2404-10
11948153
Appl Environ Microbiol. 1987 Jan;53(1):208-10
16347259
J Biol Chem. 1976 Nov 25;251(22):6994-7000
11214
Arch Biochem Biophys. 2008 May 1;473(1):61-8
18329375
Phytochemistry. 2001 Nov;58(5):739-42
11672738
Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2702-7
11867731
J Bacteriol. 1978 Jan;133(1):53-8
22538
Biochemistry. 2006 Nov 21;45(46):13889-98
17105207
Biochem J. 1999 Jun 1;340 ( Pt 2):491-5
10333494
Eur J Biochem. 2002 Dec;269(24):6302-7
12473127
Nitric Oxide. 2001 Feb;5(1):62-71
11178938
J Bacteriol. 1979 Feb;137(2):916-24
33965
Proc Natl Acad Sci U S A. 2005 Apr 12;102(15):5547-51
15802471
J Gen Microbiol. 1991 Aug;137(8):2025-32
1720168
J Biol Chem. 1987 Oct 25;262(30):14428-34
3667582
J Biol Chem. 2003 Dec 5;278(49):48674-83
14506256
Arch Biochem Biophys. 2010 Jan 1;493(1):53-61
19577534
Appl Environ Microbiol. 2007 Jun;73(12):3850-8
17449687
J Biol Chem. 1994 Sep 9;269(36):22459-62
8077188
Proc Natl Acad Sci U S A. 1977 Sep;74(9):3767-71
269430
Nat Chem Biol. 2009 Aug;5(8):521-5
19620987
Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
9254694
Biochem J. 1965 Sep;96(3):766-70
4285894
J Biol Chem. 2005 Feb 18;280(7):5195-204
15582992
Genome Res. 1999 Sep;9(9):868-77
10508846
J Biol Chem. 1964 May;239:1509-14
14189885
Cell Mol Life Sci. 2006 Mar;63(5):517-23
16465439
J Biol Chem. 1961 Mar;236:781-90
13712439
J Biol Chem. 1960 Mar;235:589-94
13846369
J Biol Chem. 1977 Jun 25;252(12):4361-70
16930