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MetaCyc Enzyme: 2-hydroxybiphenyl-3-monooxygenase [multifunctional]

Gene: hbpA Accession Number: G-15026 (MetaCyc)

Synonyms: G-15027

Species: Pseudomonas nitroreducens

Subunit composition of 2-hydroxybiphenyl-3-monooxygenase [multifunctional] = [HbpA]4
         2-hydroxybiphenyl-3-monooxygenase subunit = HbpA

Summary:
The native apparent molecular mass was determined by gel filtration chromatography and crosslinking studies [Suske97].

This novel, FAD-containing monooxygenase catalyzes the NADH-dependent ortho hydroxylation of 2-hydroxybiphenyl to biphenyl-2, 3-diol (2,3-dihydroxybiphenyl) ([Kohler88, Kohler93, Suske97] and in [Jaspers01]). This aromatic hydroxylase has a broad substrate range [Kohler88]. It can catalyze the regioselective ortho-hydroxylation at the C3-position of a variety of 2-substituted phenols to the corresponding catechols. This makes it potentially useful in the biocatalytic synthesis of 3-substituted catechols, which are difficult to synthesize chemically (in [Held99]). Directed evolution approaches have been used to improve and expand its catalytic properties [Meyer02, Meyer02a, Meyer03].

Although this enzyme can also use 2-propylphenol and 2-isopropylphenol as substrates, growth of wild-type Pseudomonas nitroreducens with 2-propylphenol does not induce it. However, the enzyme can be induced in the regulatory mutant Pseudomonas nitroreducens Prp by growth on 2-propylphenol, allowing the organism to utilize 2-propylphenol or 2-isopropylphenol as sole source of carbon and energy (see pathways 2-propylphenol degradation and 2-isopropylphenol degradation) [Kohler93, Reichlin94].

Kinetic studies of the recombinant enzyme led to a proposal for a reaction mechanism that involves a ternary complex [Suske99]. Recombinant enzyme has been crystallized and subjected to preliminary X-ray analysis in both its native and seleno-L-methionine-labeled state [Meyer03a].
The subunit apparent molecular mass was determined by SDS-PAGE. Each subunit contained one molecule of noncovalently bound FAD [Suske97].

Molecular Weight of Polypeptide: 63.761 kD (from nucleotide sequence), 60.0 kD (experimental) [Suske97 ]

Molecular Weight of Multimer: 256.0 kD (experimental) [Suske97]

pI: 6.3 [Suske97]

Unification Links: Protein Model Portal:O06647 , UniProt:O06647

Relationship Links: Entrez-Nucleotide:PART-OF:U73900 , InterPro:IN-FAMILY:IPR002938 , InterPro:IN-FAMILY:IPR003042 , Pfam:IN-FAMILY:PF01494 , Prints:IN-FAMILY:PR00420

Gene-Reaction Schematic: ?

Credits:
Created 08-Mar-2012 by Fulcher CA , SRI International


Enzymatic reaction of: 2-propylphenol monooxygenase (2-hydroxybiphenyl-3-monooxygenase [multifunctional])

EC Number: 1.14.13.-

2-propylphenol + NADH + oxygen + H+ <=> 3-propylcatechol + NAD+ + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction in which it was curated.

The reaction is physiologically favored in the direction shown.

In Pathways: 2-propylphenol degradation

Summary:
See the summary below for substrate 2-hydroxybiphenyl.

Cofactors or Prosthetic Groups: FAD [Suske97]


Enzymatic reaction of: 2-isopropylphenol monooxygenase (2-hydroxybiphenyl-3-monooxygenase [multifunctional])

EC Number: 1.14.13.-

2-isopropylphenol + NADH + oxygen + H+ <=> 3-isopropylcatechol + NAD+ + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction in which it was curated.

The reaction is physiologically favored in the direction shown.

In Pathways: 2-isopropylphenol degradation

Summary:
See the summary below for substrate 2-hydroxybiphenyl.

Cofactors or Prosthetic Groups: FAD [Suske97]


Enzymatic reaction of: 2,2',3-trihydroxybiphenyl monooxygenase (2-hydroxybiphenyl-3-monooxygenase [multifunctional])

EC Number: 1.14.13.-

2,2',3-trihydroxybiphenyl + NADH + oxygen + H+ <=> 2,2',3,3'-tetrahydroxybiphenyl + NAD+ + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction in which it was curated.

The reaction is physiologically favored in the direction shown.

In Pathways: 2,2'-dihydroxybiphenyl degradation

Summary:
See the summary below for substrate 2-hydroxybiphenyl.

Cofactors or Prosthetic Groups: FAD [Suske97]


Enzymatic reaction of: 2,2'-hydroxybiphenyl monooxygenase (2-hydroxybiphenyl-3-monooxygenase [multifunctional])

EC Number: 1.14.13.44

2,2'-dihydroxybiphenyl + NADH + oxygen + H+ <=> 2,2',3-trihydroxybiphenyl + NAD+ + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is physiologically favored in the direction shown.

In Pathways: 2,2'-dihydroxybiphenyl degradation

Summary:
See the summary below for substrate 2-hydroxybiphenyl.

Cofactors or Prosthetic Groups: FAD [Suske97]


Enzymatic reaction of: 2-hydroxybiphenyl-3-monooxygenase

EC Number: 1.14.13.44

2-hydroxybiphenyl + NADH + H+ + oxygen <=> biphenyl-2, 3-diol + NAD+ + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is physiologically favored in the direction shown.

Alternative Substrates for 2-hydroxybiphenyl: 2-sec-butylphenol [Kohler88 ] , 2-tert-butylphenol [Kohler88 ] , 2,5-dihydroxybiphenyl [Kohler88 ] , 2-ethylphenol [Kohler88 ] , 2-methylphenol [Kohler88 ] , 1-naphthol [Kohler88 ]

In Pathways: 2-hydroxybiphenyl degradation

Summary:
The apparent Km values were determined at 30 °C, pH 7.5. The biphenyl-2, 3-diol (2,3-dihydroxybiphenyl) reaction product was identified by GC-MS analysis [Suske97].

Cofactors or Prosthetic Groups: FAD [Suske97]

Inhibitors (Mixed): biphenyl-2, 3-diol [Suske97]

Inhibitors (Unknown Mechanism): Cu2+ [Suske97] , Ag+ [Suske97] , Hg2+ [Suske97] , chloride [Suske97] , p-hydroxymercuribenzoate [Suske97]

Kinetic Parameters:

Substrate
Km (μM)
Citations
oxygen
29.2
[Suske97]
2-hydroxybiphenyl
2.8
[Suske97]
NADH
26.8
[Suske97]

T(opt): 33 °C [Suske97]

pH(opt): 7.5 [Suske97]

History:
3/12/2012 (fulcher) Merged genes G-15027/hbpA into G-15026/hbpA


References

Held99: Held M, Schmid A, Kohler HP, Suske W, Witholt B, Wubbolts MG (1999). "An integrated process for the production of toxic catechols from toxic phenols based on a designer biocatalyst." Biotechnol Bioeng 62(6);641-8. PMID: 9951522

Jaspers01: Jaspers MC, Schmid A, Sturme MH, Goslings DA, Kohler HP, Roelof Van Der Meer J (2001). "Transcriptional organization and dynamic expression of the hbpCAD genes, which encode the first three enzymes for 2-hydroxybiphenyl degradation in Pseudomonas azelaica HBP1." J Bacteriol 183(1);270-9. PMID: 11114926

Kohler88: Kohler HP, Kohler-Staub D, Focht DD (1988). "Degradation of 2-hydroxybiphenyl and 2,2'-dihydroxybiphenyl by Pseudomonas sp. strain HBP1." Appl Environ Microbiol 54(11);2683-8. PMID: 3214154

Kohler93: Kohler HP, van der Maarel MJ, Kohler-Staub D (1993). "Selection of Pseudomonas sp. strain HBP1 Prp for metabolism of 2-propylphenol and elucidation of the degradative pathway." Appl Environ Microbiol 59(3);860-6. PMID: 8481010

Kohler93a: Kohler HP, Schmid A, van der Maarel M (1993). "Metabolism of 2,2'-dihydroxybiphenyl by Pseudomonas sp. strain HBP1: production and consumption of 2,2',3-trihydroxybiphenyl." J Bacteriol 175(6);1621-8. PMID: 8449871

Meyer02: Meyer A, Schmid A, Held M, Westphal AH, Rothlisberger M, Kohler HP, van Berkel WJ, Witholt B (2002). "Changing the substrate reactivity of 2-hydroxybiphenyl 3-monooxygenase from Pseudomonas azelaica HBP1 by directed evolution." J Biol Chem 277(7);5575-82. PMID: 11733527

Meyer02a: Meyer A, Wursten M, Schmid A, Kohler HP, Witholt B (2002). "Hydroxylation of indole by laboratory-evolved 2-hydroxybiphenyl 3-monooxygenase." J Biol Chem 277(37);34161-7. PMID: 12105208

Meyer03: Meyer A, Held M, Schmid A, Kohler HP, Witholt B (2003). "Synthesis of 3-tert-butylcatechol by an engineered monooxygenase." Biotechnol Bioeng 81(5);518-24. PMID: 12514800

Meyer03a: Meyer A, Tanner D, Schmid A, Sargent DF, Kohler HP, Witholt B (2003). "Crystallization and preliminary X-ray analysis of native and selenomethionine 2-hydroxybiphenyl 3-monooxygenase." Acta Crystallogr D Biol Crystallogr 59(Pt 4);741-3. PMID: 12657798

Reichlin94: Reichlin F, Kohler HP (1994). "Pseudomonas sp. strain HBP1 Prp degrades 2-isopropylphenol (ortho-cumenol) via meta cleavage." Appl Environ Microbiol 60(12);4587-91. PMID: 7811094

Suske97: Suske WA, Held M, Schmid A, Fleischmann T, Wubbolts MG, Kohler HP (1997). "Purification and characterization of 2-hydroxybiphenyl 3-monooxygenase, a novel NADH-dependent, FAD-containing aromatic hydroxylase from Pseudomonas azelaica HBP1." J Biol Chem 272(39);24257-65. PMID: 9305879

Suske99: Suske WA, van Berkel WJ, Kohler HP (1999). "Catalytic mechanism of 2-hydroxybiphenyl 3-monooxygenase, a flavoprotein from Pseudomonas azelaica HBP1." J Biol Chem 274(47);33355-65. PMID: 10559214


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of MetaCyc: Caspi et al, Nucleic Acids Research 42:D459-D471 2014
Page generated by SRI International Pathway Tools version 18.5 on Tue Nov 25, 2014, BIOCYC13A.