|Gene:||fadH||Accession Numbers: G36 (EcoCyc), b3081, ECK3071|
2,4-Dienoyl-CoA reductase functions in the reductive removal of double bonds extending from even-numbered carbon atoms in unsaturated and polyunsaturated fatty acids.
The product of the reaction catalyzed by E. coli FadH is 2-trans-enoyl-CoA, in contrast with the bovine enzyme, which produces 3-trans-enoyl-CoA. FadH is able to catalyze the reduction of 2-trans,4-cis and 2-trans,4-trans isomers with almost equal efficiency [Dommes84]. This is not due to a cis-trans isomerase activity of the enzyme [Liang00]. Kinetic data first suggested that the reaction proceeds via a ping-pong mechanism [He97]. A crystal structure of the enzyme has been solved at 2.2 Å resolution. A possible electron transfer pathway from the electron source, NADPH, via FAD, the 4Fe-4S cluster, and FMN was proposed, and the authors conclude that a ping-pong kinetic mechanism is unlikely [Hubbard03]. Site-directed mutagenesis was used to identify residues involved in catalysis and to support the proposed reaction mechanism [Tu08].
Expression is induced by growth on oleate and is subject to catabolite repression [Dommes84, You89]. Regulation of fadH expression was later shown to be complex and involves three independent regulators, FadR, ArcA, and cAMP-CRP [Feng10, Feng12] as well as HipB [Lin13].
|Map Position: [3,229,687 -> 3,231,705] (69.61 centisomes, 251°)||Length: 2019 bp / 672 aa|
Molecular Weight of Polypeptide: 72.678 kD (from nucleotide sequence), 73.0 kD (experimental) [Dommes84 ]
pI: 6.55 [He97]
Unification Links: ASAP:ABE-0010124 , DIP:DIP-9562N , EchoBASE:EB2582 , EcoGene:EG12723 , EcoliWiki:b3081 , ModBase:P42593 , OU-Microarray:b3081 , PortEco:fadH , PR:PRO_000022570 , Pride:P42593 , Protein Model Portal:P42593 , RefSeq:NP_417552 , RegulonDB:G36 , SMR:P42593 , String:511145.b3081 , UniProt:P42593
Relationship Links: InterPro:IN-FAMILY:IPR001155 , InterPro:IN-FAMILY:IPR013027 , InterPro:IN-FAMILY:IPR013785 , InterPro:IN-FAMILY:IPR023753 , PDB:Structure:1PS9 , Pfam:IN-FAMILY:PF00724 , Pfam:IN-FAMILY:PF07992 , Prints:IN-FAMILY:PR00368
|Biological Process:||GO:0033543 - fatty acid beta-oxidation, unsaturated, even number, reductase/isomerase pathway
GO:0055114 - oxidation-reduction process [UniProtGOA11a, GOA01a]
|Molecular Function:||GO:0008670 - 2,4-dienoyl-CoA reductase (NADPH) activity
[GOA01, You89, Dommes84]
GO:0010181 - FMN binding [GOA01a, Liang00]
GO:0051539 - 4 iron, 4 sulfur cluster binding [UniProtGOA11a, Liang00]
GO:0071949 - FAD binding [Dommes84]
GO:0003824 - catalytic activity [GOA01a]
GO:0016491 - oxidoreductase activity [UniProtGOA11a, GOA01a]
GO:0046872 - metal ion binding [UniProtGOA11a]
GO:0051536 - iron-sulfur cluster binding [UniProtGOA11a]
|Cellular Component:||GO:0005829 - cytosol [DiazMejia09]|
|MultiFun Terms:||metabolism → biosynthesis of building blocks → fatty acids and phosphatidic acid|
|metabolism → carbon utilization → fatty acids|
|Growth Medium||Growth?||T (°C)||O2||pH||Osm/L||Growth Observations|
|LB enriched||Yes||37||Aerobic||6.95||Yes [Gerdes03, Comment 1]|
|LB Lennox||Yes||37||Aerobic||7||Yes [Baba06, Comment 2]|
|M9 medium with 1% glycerol||Yes||37||Aerobic||7.2||0.35||Yes [Joyce06, Comment 3]|
|MOPS medium with 0.4% glucose||Yes||37||Aerobic||7.2||0.22||Yes [Baba06, Comment 2]|
Enzymatic reaction of: 2,4-dienoyl-CoA reductase
Synonyms: 2,4-dienoyl-CoA reductase (NADPH), 4-enoyl-CoA reductase (NADPH), trans-2,3-didehydroacyl-CoA:NADP+ 4-oxidoreductase, DCR
EC Number: 126.96.36.199
The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.
The reaction is physiologically favored in the direction shown.
The enzyme is inhibited by both substrate and NADPH at higher concentrations [Dommes84].
Primary Physiological Regulators of Enzyme Activity: NADH
|Chain||2 -> 672|
4/2/1999 (pkarp) Merged genes G7600/b3081 and G36/fadH
Markus Krummenacker on Tue Oct 14, 1997:
Gene object created from Blattner lab Genbank (v. M52) entry.
Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554
DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114
Dommes82: Dommes V, Luster W, Cvetanovic M, Kunau WH (1982). "Purification by affinity chromatography of 2,4-dienoyl-CoA reductases from bovine liver and Escherichia coli." Eur J Biochem 125(2);335-41. PMID: 6749495
Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938
He97: He XY, Yang SY, Schulz H (1997). "Cloning and expression of the fadH gene and characterization of the gene product 2,4-dienoyl coenzyme A reductase from Escherichia coli." Eur J Biochem 1997;248(2);516-20. PMID: 9346310
Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394
Kimura04: Kimura C, Mizugaki M, Yamanaka H, Fujino M, Morishima T (2004). "[2,4-Dienoyl-CoA reductases: from discovery toward pathophysiological significance]." Nihon Rinsho 62(8);1577-83. PMID: 15344554
Liang00: Liang X, Thorpe C, Schulz H (2000). "2,4-Dienoyl-CoA reductase from Escherichia coli is a novel iron-sulfur flavoprotein that functions in fatty acid beta-oxidation." Arch Biochem Biophys 2000;380(2);373-9. PMID: 10933894
Lin13: Lin CY, Awano N, Masuda H, Park JH, Inouye M (2013). "Transcriptional Repressor HipB Regulates the Multiple Promoters in Escherichia coli." J Mol Microbiol Biotechnol 23(6);440-447. PMID: 24089053
Mizugaki82: Mizugaki M, Nishimaki T, Yamamoto H, Nishimura S, Sagi M, Yamanaka H (1982). "Studies on the metabolism of unsaturated fatty acids. VIII. Induction of 2,4-dienoyl-CoA reductase in Escherichia coli on the addition of unsaturated fatty acids." J Biochem 91(4);1453-6. PMID: 7047514
Mizugaki82a: Mizugaki M, Kimura C, Nishimaki T, Yamamoto H, Sagi M, Nishimura S, Yamanaka H (1982). "Studies on the metabolism of unsaturated fatty acids. X. Purification and some properties of 2,4-dienoyl-CoA reductase from Escherichia coli." J Biochem 92(5);1671-4. PMID: 6759505
Mizugaki83: Mizugaki M, Kimura C, Nishimaki T, Kawaguchi A, Okuda S, Yamanaka H (1983). "Studies on the metabolism of unsaturated fatty acids. XII. Reaction catalyzed by 2,4-dienoyl-CoA reductase of Escherichia coli." J Biochem 94(2);409-13. PMID: 6355075
Tu08: Tu X, Hubbard PA, Kim JJ, Schulz H (2008). "Two distinct proton donors at the active site of Escherichia coli 2,4-dienoyl-CoA reductase are responsible for the formation of different products." Biochemistry 47(4);1167-75. PMID: 18171025
You89: You SY, Cosloy S, Schulz H (1989). "Evidence for the essential function of 2,4-dienoyl-coenzyme A reductase in the beta-oxidation of unsaturated fatty acids in vivo. Isolation and characterization of an Escherichia coli mutant with a defective 2,4-dienoyl-coenzyme A reductase." J Biol Chem 1989;264(28);16489-95. PMID: 2506179
©2014 SRI International, 333 Ravenswood Avenue, Menlo Park, CA 94025-3493