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Escherichia coli K-12 substr. MG1655 Enzyme: 2,3-dihydroxyphenylpropionate 1,2-dioxygenase



Gene: mhpB Accession Numbers: M011 (EcoCyc), b0348, ECK0345

Regulation Summary Diagram: ?

Subunit composition of 2,3-dihydroxyphenylpropionate 1,2-dioxygenase = [MhpB]4
         3-(2,3-dihydroxyphenyl)propionate dioxygenase monomer = MhpB

Summary:
3-(2,3-dihydroxyphenyl)propionate dioxygenase (MhpB) is a non-heme-iron(II)-dependent extradiol catechol dioxygenase; it catalyzes the meta cleavage of the phenolic ring structure of 3-(2,3-dihydroxyphenyl)propionate. This compound is a common catabolic intermediate of both 3-phenylpropionate and 3-(3-hydroxyphenyl)propionate degradation [Burlingame83, Bugg93].

The catalytic mechanism of MhpB is based on acid-base catalysis involving the His115 and His179 residues [Mendel04] assisting direct 1,2-alkenyl migration to form a lactone intermediate [Xin08]. A H115Y mutant catalyzes lactonization of the substrate [Mendel05]. Directed evolution of MhpB yields enzymes that are able to catalyze the intradiol catechol dioxygenase reaction [Schlosrich06]. MhpB can also catalyze the formation of a 2-tropolone ring-expansion product through a pinacol-type rearrangement [Xin10].

An mhpB mutant does not grow with m-hydroxyphenylpropionate (MHP) or hydroxycinnamate (HCA) as the sole source of carbon [Burlingame86].

Citations: [Ferrandez97]

Gene Citations: [Ferrandez97]

Map Position: [369,501 -> 370,445] (7.96 centisomes)
Length: 945 bp / 314 aa

Molecular Weight of Polypeptide: 34.196 kD (from nucleotide sequence), 36.0 kD (experimental) [Bugg93 ]

Molecular Weight of Multimer: 134.0 kD (experimental) [Bugg93]

Unification Links: ASAP:ABE-0001198 , EchoBASE:EB4167 , EcoGene:EG20274 , EcoliWiki:b0348 , ModBase:P0ABR9 , OU-Microarray:b0348 , PortEco:mhpB , PR:PRO_000023229 , Protein Model Portal:P0ABR9 , RefSeq:NP_414882 , RegulonDB:M011 , SMR:P0ABR9 , String:511145.b0348 , UniProt:P0ABR9

Relationship Links: InterPro:IN-FAMILY:IPR004183 , InterPro:IN-FAMILY:IPR023789 , Pfam:IN-FAMILY:PF02900

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0019622 - 3-(3-hydroxy)phenylpropionate catabolic process Inferred from experiment [Burlingame86]
GO:0046271 - phenylpropanoid catabolic process Inferred from experiment [Burlingame86]
GO:0006725 - cellular aromatic compound metabolic process Inferred by computational analysis [GOA01]
GO:0019380 - 3-phenylpropionate catabolic process Inferred by computational analysis [UniProtGOA12]
GO:0019439 - aromatic compound catabolic process Inferred by computational analysis [UniProtGOA11, GOA06, GOA01]
GO:0055114 - oxidation-reduction process Inferred by computational analysis [UniProtGOA11, GOA01]
Molecular Function: GO:0008198 - ferrous iron binding Inferred from experiment Inferred by computational analysis [GOA01, Bugg93]
GO:0047070 - 3-carboxyethylcatechol 2,3-dioxygenase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Bugg93]
GO:0005506 - iron ion binding Inferred by computational analysis [GOA06, GOA01]
GO:0016491 - oxidoreductase activity Inferred by computational analysis [UniProtGOA11, GOA01]
GO:0051213 - dioxygenase activity Inferred by computational analysis [UniProtGOA11]

MultiFun Terms: metabolism carbon utilization fatty acids

Essentiality data for mhpB knockouts: ?

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]
Yes [Feist07, Comment 4]

Credits:
Last-Curated ? 26-Oct-2011 by Keseler I , SRI International


Enzymatic reaction of: 2,3-dihydroxycinnamate dioxygenase (2,3-dihydroxyphenylpropionate 1,2-dioxygenase)

EC Number: 1.13.11.16

2,3-dihydroxy-trans-cinnamate + oxygen <=> (2Z,4E,7E)-2-hydroxy-6-oxonona-2,4,7-triene-1,9-dioate + H+

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.

In Pathways: cinnamate and 3-hydroxycinnamate degradation to 2-oxopent-4-enoate

Kinetic Parameters:

Substrate
Km (μM)
Citations
2,3-dihydroxy-trans-cinnamate
36.0
[Spence96]


Enzymatic reaction of: 3-(2,3-dihydroxyphenyl)propionate 1,2-dioxygenase (2,3-dihydroxyphenylpropionate 1,2-dioxygenase)

Synonyms: 2,3-dihydroxyphenylpropionate 1,2-dioxygenase, 3-(2,3-dihydroxyphenyl)propionate dioxygenase

EC Number: 1.13.11.16

3-(2,3-dihydroxyphenyl)propanoate + oxygen <=> (2Z,4E)-2-hydroxy-6-oxonona-2,4-diene-1,9-dioate + H+

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.

Alternative Substrates for 3-(2,3-dihydroxyphenyl)propanoate: 3-ethylcatechol [Spence96 ] , 3-propylcatechol [Spence96 ] , methyl-2,3-dihydroxyphenylpropionate [Spence96 ] , 3-phenylethylcatechol [Spence96 ] , 3-methylcatechol [Bugg93 ] , catechol [Bugg93 ]

In Pathways: 3-phenylpropionate and 3-(3-hydroxyphenyl)propionate degradation , 3-phenylpropanoate and 3-(3-hydroxyphenyl)propanoate degradation to 2-oxopent-4-enoate

Cofactors or Prosthetic Groups: Fe2+ [Comment 5, Bugg93]

Inhibitors (Unknown Mechanism): 5,5'-dithio-bis-2-nitrobenzoate [Spence96] , p-hydroxymercuribenzoate [Spence96]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
catechol
720.0
[Bugg93]
3-methylcatechol
90.0
[Bugg93]
3-phenylethylcatechol
94.0
[Spence96]
methyl-2,3-dihydroxyphenylpropionate
37.0
[Spence96]
3-propylcatechol
154.0
[Spence96]
3-ethylcatechol
185.0
[Spence96]
3-(2,3-dihydroxyphenyl)propanoate
26.0
29.0
[Bugg93]

pH(opt): 8-9.5 [Bugg93]


Sequence Features

Feature Class Location Citations Comment
Mutagenesis-Variant 114
[Mendel04, UniProt11]
Alternate sequence: D → N; UniProt: Low level of catalytic activity, 600-fold lower than the wild-type enzyme. More than 8000-fold decrease in affinity.
Alternate sequence: D → A; UniProt: Complete loss of extradiol cleavage activity.
Mutagenesis-Variant 115
[Mendel04, UniProt11]
Alternate sequence: H → Y; UniProt: Complete loss of activity.
Alternate sequence: H → Q; UniProt: Complete loss of activity.
Alternate sequence: H → A; UniProt: Complete loss of extradiol cleavage activity.
Active-Site 115
[UniProt10a]
UniProt: Proton donor;
Sequence-Conflict 138 -> 140
[Spence96, UniProt10a]
Alternate sequence: ING → NKA; UniProt: (in Ref. 1; BAA13053);
Sequence-Conflict 152
[Spence96, UniProt10a]
Alternate sequence: R → H; UniProt: (in Ref. 1; BAA13053);
Sequence-Conflict 157
[Spence96, UniProt10a]
Alternate sequence: A → T; UniProt: (in Ref. 1; BAA13053);
Mutagenesis-Variant 179
[Mendel04, UniProt11]
Alternate sequence: H → Y; UniProt: Complete loss of activity.
Alternate sequence: H → Q; UniProt: Complete loss of activity.
Alternate sequence: H → A; UniProt: Complete loss of activity.
Active-Site 179
[UniProt10a]
UniProt: Proton acceptor;
Mutagenesis-Variant 181
[Mendel04, UniProt11]
Alternate sequence: P → H; UniProt: More than 60-fold decrease in catalytic activity and affinity.
Alternate sequence: P → A; UniProt: More than 2-fold decrease in catalytic activity and 100-fold decrease in affinity.


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b0348 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene M011.


References

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

Bugg93: Bugg TD (1993). "Overproduction, purification and properties of 2,3-dihydroxyphenylpropionate 1,2-dioxygenase from Escherichia coli." Biochim Biophys Acta 1993;1202(2);258-64. PMID: 8399388

Burlingame83: Burlingame R, Chapman PJ (1983). "Catabolism of phenylpropionic acid and its 3-hydroxy derivative by Escherichia coli." J Bacteriol 1983;155(1);113-21. PMID: 6345502

Burlingame86: Burlingame RP, Wyman L, Chapman PJ (1986). "Isolation and characterization of Escherichia coli mutants defective for phenylpropionate degradation." J Bacteriol 1986;168(1);55-64. PMID: 3531186

Diaz98: Diaz E, Ferrandez A, Garcia JL (1998). "Characterization of the hca cluster encoding the dioxygenolytic pathway for initial catabolism of 3-phenylpropionic acid in Escherichia coli K-12." J Bacteriol 1998;180(11);2915-23. PMID: 9603882

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Ferrandez97: Ferrandez A, Garcia JL, Diaz E (1997). "Genetic characterization and expression in heterologous hosts of the 3-(3-hydroxyphenyl)propionate catabolic pathway of Escherichia coli K-12." J Bacteriol 1997;179(8);2573-81. PMID: 9098055

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

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

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

Mendel04: Mendel S, Arndt A, Bugg TD (2004). "Acid-base catalysis in the extradiol catechol dioxygenase reaction mechanism: site-directed mutagenesis of His-115 and His-179 in Escherichia coli 2,3-dihydroxyphenylpropionate 1,2-dioxygenase (MhpB)." Biochemistry 43(42);13390-6. PMID: 15491145

Mendel05: Mendel S, Arndt A, Bugg TD (2005). "Lactone synthesis activity in a site-directed mutant of an extradiol catechol dioxygenase enzyme." Chem Commun (Camb) (5);666-8. PMID: 15672171

Schlosrich06: Schlosrich J, Eley KL, Crowley PJ, Bugg TD (2006). "Directed evolution of a non-heme-iron-dependent extradiol catechol dioxygenase: identification of mutants with intradiol oxidative cleavage activity." Chembiochem 7(12);1899-908. PMID: 17051653

Spence96: Spence EL, Kawamukai M, Sanvoisin J, Braven H, Bugg TD (1996). "Catechol dioxygenases from Escherichia coli (MhpB) and Alcaligenes eutrophus (MpcI): sequence analysis and biochemical properties of a third family of extradiol dioxygenases." J Bacteriol 1996;178(17);5249-56. PMID: 8752345

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Xin08: Xin M, Bugg TD (2008). "Evidence from mechanistic probes for distinct hydroperoxide rearrangement mechanisms in the intradiol and extradiol catechol dioxygenases." J Am Chem Soc 130(31);10422-30. PMID: 18627158

Xin10: Xin M, Bugg TD (2010). "Biomimetic formation of 2-tropolones by dioxygenase-catalysed ring expansion of substituted 2,4-cyclohexadienones." Chembiochem 11(2);272-6. PMID: 20013980

Other References Related to Gene Regulation

Torres03: Torres B, Porras G, Garcia JL, Diaz E (2003). "Regulation of the mhp cluster responsible for 3-(3-hydroxyphenyl) propionic acid degradation in escherichia coli." J Biol Chem 278(30);27575-85. PMID: 12748194


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Sun Nov 23, 2014, BIOCYC14A.