Definition of 4-hydroxybenzoate catabolism
As rules and steps, or see full text
Rules
Overview: 4-hydroxybenzoate catabolism in GapMind is based on aerobic oxidation to 3,4-hydroxybenzoate (protocatechuate), followed by meta, ortho, or para cleavage; or reduction to benzoyl-CoA (part of a MetaCyc pathway for phenol degradation, link)
- all:
- 4-hydroxybenzoate-transport, pobA and protocatechuate-degradation
- or 4-hydroxybenzoate-transport, hcl, hcrA, hcrB, hcrC and benzoyl-CoA-degradation
- Comment: An aerobic route for degradation of 4-hydroxybenzoate involves 4-hydroxybenzoate 3-monooxygenase pobA, which forms protocatechuate (3,4-dihydroxybenzoate). Alternatively, 4-hydroxybenzoate can be activated to 4-hydroxybenzoyl-CoA by hcl and reduced to benzoyl-CoA by hcrABC (link).
- protocatechuate-degradation:
- ligA, ligB, ligC, ligI, ligU, ligJ and ligK
- or pcaH, pcaG, pcaB, pcaC, pcaD and 3-oxoadipate-degradation
- or praA and 2-hydroxymuconate-6-semialdehyde-degradation
- Comment: In the meta-cleavage pathway (link), the 4,5-dioxygenase ligAB splits protocatechuate to 4-carboxy-2-hydroxymuconate-6-semialdehyde. (In solution, this is in the hemiacetal form.) The semialdehyde is oxidized to 2-pyrone-4,6-dicarboxylate, hydrolyzed to (1E)-4-oxobut-1-ene-1,2,4-tricarboxylate, tautomerized to (1E,3E)-4-hydroxybuta-1,3-diene-1,2,4-tricarboxylate, hydrated to 2-hydroxy-4-oxobutane-1,2,4-tricarboxylate (4-oxalocitramalate), and split by an aldolase to pyruvate and oxaloacetate. In the ortho-cleavage pathway (link), the 3,4-oxygenase pcaHG cleaves the ring to 3-carboxy-cis,cis-muconate, a cycloisomerase forms 4-carboxymuconolactone (2-carboxy-2,5-dihydro-5-oxofuran-2-yl)-acetate), a decarboxylase forms 3-oxoadipate enol lactone ((4,5-dihydro-5-oxofuran-2-yl)-acetate), and a hydrolase forms 3-oxoadipate. In the para-cleavage pathway (link), the 2,3-dioxygenase praA forms (2Z,4Z)-2-hydroxy-5-carboxymuconate-6-semialdehyde, which spontaneously decarboxylates to (2Z,4E)-2-hydroxy-6-oxohexa-2,4-dienoate, also known as 2-hydroxymuconate 6-semialdehyde.
- 2-hydroxymuconate-6-semialdehyde-degradation:
- praB, praC, praD and 2-hydroxypenta-2,4-dienoate-degradation
- or xylF and 2-hydroxypenta-2,4-dienoate-degradation
- Comment: Dehydrogenase praB forms 2-hydroxymuconate, tautomerase praC forms (3E)-2-oxohex-3-enedioate (2-oxalocrotonate), and decarboxylase praD yields 2-hydroxypenta-2,4-dienoate (HPD). (This series of steps is part of protocatechuate para-cleavage, link, or catechol degradation II, link.) Or, hydrolase xylF forms HPD and formate. (This is part of a MetaCyc pathway for catechol degradation, link.)
- benzoyl-CoA-degradation:
- benzoyl-CoA-reductase, dch, had, oah, pimB and glutaryl-CoA-degradation
- or benzoyl-CoA-reductase, Ch1CoA, badK, badH, badI, pimD, pimC, pimF and glutaryl-CoA-degradation
- or boxA, boxB, boxC, boxD, paaF, paaH and paaJ2
- Comment: Benzoyl-CoA can be degraded anaerobically (link) by reduction to cyclohex-1,5-diene-1-carbonyl-CoA, followed by hydratase (dch) to 6-hydroxycyclohex-1-ene-1-carbonyl-CoA, a dehydrogenase to 6-oxocyclohex-1-ene-1-carbonyl-CoA, a hydrolase to 2-hydroxy-6-oxocycloheane-1-carbonyl-CoA, a ring-opening hydrolase to 3-hydroxypimeloyl-CoA [the last two steps are both catalyzed by oah], a dehydrogenase to 3-oxopimeloyl-CoA [not linked to sequence and omitted], and an acetyltransferase to glutaryl-CoA and acetyl-CoA. Alternatively, after reduction to cyclohex-1,5-diene-1-carbonyl-CoA, Ch1CoA can further reduce it to cyclohex-1-ene-1-carboxyl-CoA (link), followed by hydration to 2-hydroxy-cyclohexane-1-carbonyl-CoA, oxidation to 2-ketocyclohexane-1-carbonyl-CoA, cleavage by a ring-opening hydrolase to pimeloyl-CoA, oxidation to 2,3-didehydropimeloyl-CoA, hydration to 3-hydroxypimeloyl-C, oxidation to 3-oxopimeloyl-CoA and cleavage by a thiolase to glutaryl-CoA and acetyl-CoA. Benzoyl-CoA degradation can be degraded aerobically (link) by an epoxidase (boxAB) that forms 2,3-epoxy-2-3-dihydrobenzoyl-CoA; a dihydrolase forms cis-3,4-dihydroadipyl-CoA semialdehyde and formate; a dehydrogenase forms cis-3,4-dehydroadipyl-CoA; and an unknown isomerase forms trans-2,3-dehydroadipyl-CoA. This is converted to succinyl-CoA as in the anaerobic pathway (paaF, paaH, and paaJ2).
- glutaryl-CoA-degradation: gcdH, ech, fadB and atoB
- Comment: In MetaCyc pathway glutaryl-CoA degradation (link), glutaryl-CoA is oxidized to (E)-glutaconyl-CoA and oxidatively decarboxylated to crotonyl-CoA (both by the same enzyme), hydrated to 3-hydroxybutanoyl-CoA, oxidized to acetoacetyl-CoA, and cleaved to two acetyl-CoA.
- benzoyl-CoA-reductase:
- bcrA, bcrB, bcrC and bcrD
- or bamB, bamC, bamD, bamE, bamF, bamG, bamH and bamI
- Comment: Benzoyl-CoA reduction is energetically unfavorable. There are two classes of reductases: class I enzymes (bcrABCD) use ATP to drive the reaction, while class II enzymes (bamBCDEFGHI) are thought to us an electron bifurcation. SYN_02587 (Q2LQN9) from Syntrophus aciditrophicus, which can oxidize cyclohex-1,5-diene-1-carbonyl-CoA to benzoyl-CoA, is not included because it seems to lack a mechanism to drive benzoyl-CoA reduction.
- 2-hydroxypenta-2,4-dienoate-degradation: mhpD, mhpE and acetaldehyde-degradation
- Comment: (2Z)-2-hydroxypenta-2,4-dienoate (HPD) is a common intermediate in the aerobic degradation of many aromatic compounds. In MetaCyc pathway 2-hydroxypenta-2,4-dienoate degradation (link), HPD is hydrated to (S)-4-hydroxy-2-oxopentanoate and an aldolase cleaves it to pyruvate and acetaldehyde.
- acetaldehyde-degradation:
- ald-dh-CoA
- or adh and acs
- or adh, ackA and pta
- Comment: Acetaldehyde can be oxidized to acetyl-CoA, or oxidized to acetate and activated to acetyl-CoA by either acetyl-CoA synthetase (acs) or by acetate kinase (ackA) and phosphate acetyltransferase (pta).
- 3-oxoadipate-degradation: 3-oxodipate-CoA-transferase and pcaF
- Comment: MetaCyc pathway 3-oxoadipate degradation (link) involves activation by CoA (using succinyl-CoA) and a thiolase (succinyltransferase) reaction that splits it to acetyl-CoA and succinyl-CoA.
- 3-oxodipate-CoA-transferase:
- pcaI and pcaJ
- or catI and catJ
- Comment: Two different types of 3-oxoadipate CoA-transferases (EC 2.8.3.6) are known. They are both heteromeric with each subunit containing a CoA-transferase domain
- 4-hydroxybenzoate-transport:
- pcaK
- or fcbT1, fcbT2 and fcbT3
- Comment: Transporters were identified using: query: transporter:4-hydroxybenzoate:p-hydroxybenzoate:4-hydroxybenzoic acid:p-hydroxybenzoic acid
Steps
pcaK: 4-hydroxybenzoate transporter pcaK
- Curated sequence Q43975: 4-hydroxybenzoate transporter PcaK; 4-HB transporter
- Curated sequence Q51955: 4-hydroxybenzoate transporter PcaK. 4-Hydroxybenzoate/protocatechuate porter
- Total: 2 characterized proteins
fcbT1: tripartite 4-hydroxybenzoate transporter, substrate-binding component FcbT1
- Curated sequence Q9RBR1: FcbT1, component of Tripartite 4-chlorobenzoate symporter (also binds and may transport 4-bromo-, 4-iodo-, and 4-fluorobenzoate and with a lower affinity, 3-chlorobenzoate, 2-chlorobenzoate, 4-hydroxybenzoate, 3-hydroxybenzoate, and benzoate)
- Comment: The primary substrate for this system is 4-chlorobenzoate, but TCDB reports that binding of that substrate to fcbT1 is inhibited by 1 mM 4-hydroxybenzoate, suggesting a reasonable affinity for 4-hydroxybenzoate as well.
- Total: 1 characterized proteins
fcbT2: tripartite 4-hydroxybenzoate transporter, small DctQ-like component FcbT2
- Curated sequence Q9RBR0: FcbT2, component of Tripartite 4-chlorobenzoate symporter (also binds and may transport 4-bromo-, 4-iodo-, and 4-fluorobenzoate and with a lower affinity, 3-chlorobenzoate, 2-chlorobenzoate, 4-hydroxybenzoate, 3-hydroxybenzoate, and benzoate)
- Total: 1 characterized proteins
fcbT3: tripartite 4-hydroxybenzoate transporter, large permease subunit FcbT3
- Curated sequence Q9RBQ9: FcbT3, component of Tripartite 4-chlorobenzoate symporter (also binds and may transport 4-bromo-, 4-iodo-, and 4-fluorobenzoate and with a lower affinity, 3-chlorobenzoate, 2-chlorobenzoate, 4-hydroxybenzoate, 3-hydroxybenzoate, and benzoate)
- Total: 1 characterized proteins
pcaI: 3-oxoadipate CoA-transferase subunit A (PcaI)
- Curated sequence CH_021928: 3-oxoadipate CoA-transferase subunit A; EC 2.8.3.6. 3-oxoadipate CoA-transferase (subunit 2/2) (EC 2.8.3.6). α subunit of β-ketoadipate succinyl-CoA transferase (EC 2.8.3.6)
- Ignore hits to items matching 2.8.3.6 when looking for 'other' hits
- Ignore hits to items matching 2.8.3.5 when looking for 'other' hits
- Comment: Ignore EC 2.8.3.5, 3-oxoacid CoA-transferase, usually active on shorter substrates but who knows?
- Total: 1 characterized proteins
pcaJ: 3-oxoadipate CoA-transferase subunit B (PcaJ)
- Curated sequence P0A102: 3-oxoadipate CoA-transferase (subunit 1/2) (EC 2.8.3.6). 3-oxoadipate CoA-transferase subunit B; EC 2.8.3.6. β subunit of β-ketoadipate succinyl-CoA transferase (EC 2.8.3.6)
- Ignore hits to items matching 2.8.3.6 when looking for 'other' hits
- Ignore hits to items matching 2.8.3.5 when looking for 'other' hits
- Total: 1 characterized proteins
catI: 3-oxoadipate CoA-transferase subunit A (CatI)
- Curated sequence Q8VPF3: 3-oxoadipate CoA-transferase subunit A; 3-oxoadipate:succinyl-CoA transferase subunit A; Beta-ketoadipate:succinyl-CoA transferase subunit A; EC 2.8.3.6. 3-oxoadipate CoA-transferase (subunit 2/2) (EC 2.8.3.6)
- Curated sequence GFF1318: 3-oxoadipate CoA-transferase (EC 2.8.3.6)
- Curated sequence PfGW456L13_4592: 3-oxoadipate CoA-transferase (EC 2.8.3.6)
- Curated sequence AO353_17195: 3-oxoadipate CoA-transferase (EC 2.8.3.6)
- Ignore hits to items matching 2.8.3.6 when looking for 'other' hits
- Ignore hits to items matching 2.8.3.5 when looking for 'other' hits
- Comment: Ignore EC 2.8.3.5, 3-oxoacid CoA-transferase, usually active on shorter substrates but who knows?
- Total: 4 characterized proteins
catJ: 3-oxoadipate CoA-transferase subunit B (CatJ)
- Curated sequence Q8VPF2: 3-oxoadipate CoA-transferase subunit B; 3-oxoadipate:succinyl-CoA transferase subunit B; Beta-ketoadipate:succinyl-CoA transferase subunit B; EC 2.8.3.6. 3-oxoadipate CoA-transferase (subunit 1/2) (EC 2.8.3.6)
- Curated sequence GFF1319: 3-oxoadipate CoA-transferase (EC 2.8.3.6)
- Curated sequence PfGW456L13_4591: 3-oxoadipate CoA-transferase (EC 2.8.3.6)
- Curated sequence AO353_17200: 3-oxoadipate CoA-transferase (EC 2.8.3.6)
- Ignore hits to items matching 2.8.3.6 when looking for 'other' hits
- Ignore hits to items matching 2.8.3.5 when looking for 'other' hits
- Total: 4 characterized proteins
pcaF: succinyl-CoA:acetyl-CoA C-succinyltransferase
mhpD: 2-hydroxypentadienoate hydratase
mhpE: 4-hydroxy-2-oxovalerate aldolase
ald-dh-CoA: acetaldehyde dehydrogenase, acylating
- Curated proteins or TIGRFams with EC 1.2.1.10
- Ignore hits to items matching 1.1.1.1 when looking for 'other' hits
- Ignore hits to items matching 1.1.1.71 when looking for 'other' hits
- Ignore hits to items matching 1.2.1.57 when looking for 'other' hits
- Ignore hits to Q2XQZ7 when looking for 'other' hits (4-hydroxy-2-oxovalerate aldolase (EC 4.1.3.39))
- Comment: Many enzymes are multifunctional alcohol/acetaldehyde dehydrogenases, and many close homologs have just one annotation. EC 1.2.1.57 is acylating butanal dehydrogenase, which may also act on acetaldehyde. Q2XQZ7 is probably misannotated.
- Total: 2 HMMs and 20 characterized proteins
adh: acetaldehyde dehydrogenase (not acylating)
acs: acetyl-CoA synthetase, AMP-forming
ackA: acetate kinase
pta: phosphate acetyltransferase
- Curated proteins or TIGRFams with EC 2.3.1.8
- Ignore hits to P32796 when looking for 'other' hits (carnitine O-acetyltransferase (EC 2.3.1.7); phosphate acetyltransferase (EC 2.3.1.8). Carnitine O-acetyltransferase, mitochondrial; Carnitine acetylase; EC 2.3.1.7)
- Comment: BRENDA misannotates yeast's carnitine acetyltransferase with EC 2.3.1.8
- Total: 1 HMMs and 18 characterized proteins
atoB: acetyl-CoA C-acetyltransferase
- Curated proteins or TIGRFams with EC 2.3.1.9
- Ignore hits to items matching 2.3.1.16 when looking for 'other' hits
- Ignore hits to P07256 when looking for 'other' hits (acetyl-CoA C-acetyltransferase (EC 2.3.1.9). Cytochrome b-c1 complex subunit 1, mitochondrial; Complex III subunit 1; Core protein I; Ubiquinol-cytochrome c oxidoreductase core protein 1; Ubiquinol-cytochrome c reductase 44 kDa protein)
- Ignore hits to I3R3D0 when looking for 'other' hits (acetyl-CoA C-acetyltransferase (subunit 1/2) (EC 2.3.1.9))
- Ignore hits to I3RA71 when looking for 'other' hits (acetyl-CoA C-acetyltransferase (subunit 1/2) (EC 2.3.1.9))
- Ignore hits to items matching similar to acetyl-CoA acetyltransferase when looking for 'other' hits
- Comment: Produces two acetyl-CoA from acetoacetyl-CoA and CoA. EC 2.3.1.16 describes a broader range of beta-ketothiolases. This enzyme is usually homomeric, but I3R3D0 and I3RA71 are non-catalytic subunits of an enzyme from Haloferax mediterranei that also contains a "normal" catalytic subunit (I3R3D1, I3RA72). Inclusion of P07256 was an error in BRENDA. And CharProtDB includes an odd annotation of the form "similar to acetyl-CoA acetyltransferase"
- Total: 36 characterized proteins
bcrA: ATP-dependent benzoyl-CoA reductase, alpha subunit
- Curated sequence O87876: Benzoyl-CoA reductase subunit A; 3-hydroxybenzoyl-CoA reductase subunit alpha; EC 1.3.7.8; EC 1.3.99.n1. benzoyl-CoA reductase (EC 1.3.7.8). benzoyl-CoA reductase α subunit (EC 1.3.7.8)
- Curated sequence O07462: benzoyl-CoA reductase (EC 1.3.7.8). BadF (EC 1.3.7.8)
- Curated sequence Q8VUG0: benzoyl-CoA reductase (EC 1.3.7.8)
- Ignore hits to items matching 1.3.7.8 when looking for 'other' hits
- Comment: Thauera aromatica has BrcABCD; a similar system in Rhodopseudomonas palustris is known as badFEDG; and a similar system in Azoarcus is known as BzdQONP (see PMC516837 and Genbank AF521665). [The curated entries for Azoarcus, in BRENDA, are from another strain and are not quite identical to the protein sequences in AF521665]
- Total: 3 characterized proteins
bcrB: ATP-dependent benzoyl-CoA reductase, beta subunit
- Curated sequence O87875: Benzoyl-CoA reductase subunit B; 3-hydroxybenzoyl-CoA reductase subunit beta; EC 1.3.7.8; EC 1.3.99.n1. benzoyl-CoA reductase β subunit (EC 1.3.7.8)
- Curated sequence O07461: benzoyl-CoA reductase (EC 1.3.7.8). BadE (EC 1.3.7.8)
- Curated sequence Q8VUG2: benzoyl-CoA reductase (EC 1.3.7.8)
- Ignore hits to items matching 1.3.7.8 when looking for 'other' hits
- Total: 3 characterized proteins
bcrC: ATP-dependent benzoyl-CoA reductase, gamma subunit
- Curated sequence O87874: Benzoyl-CoA reductase subunit C; 3-hydroxybenzoyl-CoA reductase subunit gamma; EC 1.3.7.8; EC 1.3.99.n1. benzoyl-CoA reductase γ subunit (EC 1.3.7.8)
- Curated sequence O07460: BadD (EC 1.3.7.8). benzoyl-CoA reductase (EC 1.3.7.8)
- Curated sequence Q8VUG3: benzoyl-CoA reductase (EC 1.3.7.8)
- Ignore hits to items matching 1.3.7.8 when looking for 'other' hits
- Total: 3 characterized proteins
bcrD: ATP-dependent benzoyl-CoA reductase, delta subunit
- Curated sequence O87877: Benzoyl-CoA reductase subunit D; 3-hydroxybenzoyl-CoA reductase subunit delta; EC 1.3.7.8; EC 1.3.99.n1. benzoyl-CoA reductase δ subunit (EC 1.3.7.8)
- Curated sequence O07463: BadG (EC 1.3.7.8). benzoyl-CoA reductase (EC 1.3.7.8)
- Curated sequence Q8VUG1: benzoyl-CoA reductase (EC 1.3.7.8)
- Ignore hits to items matching 1.3.7.8 when looking for 'other' hits
- Total: 3 characterized proteins
bamB: class II benzoyl-CoA reductase, BamB subunit
- UniProt sequence Q39TV8: SubName: Full=Benzoyl-CoA reductase, putative {ECO:0000313|EMBL:ABB32316.1};
- Comment: bamBCDEFGHI has been described in Geobacter metallireducens (PMID:30674680). There is also a paper about the enzyme from Desulfocarcina cetonica but I could not find those sequences. bamB = Gmet_2087
- Total: 1 characterized proteins
bamC: class II benzoyl-CoA reductase, BamC subunit
- UniProt sequence Q39TV9: SubName: Full=Iron-sulfur cluster-binding oxidoreductase, putative benzoyl-CoA reductase electron transfer protein {ECO:0000313|EMBL:ABB32315.1};
- Comment: bamC = Gmet_2086
- Total: 1 characterized proteins
bamD: class II benzoyl-CoA reductase, BamD subunit
- UniProt sequence Q39TW0: SubName: Full=Iron-sulfur cluster-binding oxidoreductase, CCG domain pair-containing, putative benzoyl-CoA reductase electron transfer protein {ECO:0000313|EMBL:ABB32314.1};
- Comment: bamD = Gmet_2085
- Total: 1 characterized proteins
bamE: class II benzoyl-CoA reductase, BamE subunit
- UniProt sequence Q39TW1: SubName: Full=Polyferredoxin, putative benzoyl-CoA reductase electron transfer protein {ECO:0000313|EMBL:ABB32313.1};
- Comment: bamE = Gmet_2084
- Total: 1 characterized proteins
bamF: class II benzoyl-CoA reductase, BamF subunit
- UniProt sequence Q39TW2: SubName: Full=Benzoyl-CoA reductase electron transfer protein, selenocysteine-containing, putative {ECO:0000313|EMBL:ABB32312.2};
- Comment: bamF = Gmet_2083
- Total: 1 characterized proteins
bamG: class II benzoyl-CoA reductase, BamG subunit
- UniProt sequence Q39TW4: SubName: Full=Benzoyl-CoA reductase electron transfer protein, putative {ECO:0000313|EMBL:ABB32310.1};
- Comment: bamG = Gmet_2081
- Total: 1 characterized proteins
bamH: class II benzoyl-CoA reductase, BamH subunit
- UniProt sequence Q39TW5: SubName: Full=Benzoyl-CoA reductase electron transfer protein, putative {ECO:0000313|EMBL:ABB32309.1};
- Comment: bamH = Gmet_2080
- Total: 1 characterized proteins
bamI: class II benzoyl-CoA reductase, BamI subunit
- UniProt sequence Q39TW6: SubName: Full=Iron-sulfur cluster-binding protein, putative {ECO:0000313|EMBL:ABB32308.1};
- Comment: bamI = Gmet_2079
- Total: 1 characterized proteins
gcdH: glutaryl-CoA dehydrogenase
ech: (S)-3-hydroxybutanoyl-CoA hydro-lyase
- Curated proteins or TIGRFams with EC 4.2.1.150
- Ignore hits to Q97MS7 when looking for 'other' hits (short-chain-enoyl-CoA hydratase (EC 4.2.1.150))
- Curated sequence BPHYT_RS17335: trans-2,3-dehydroadipyl-CoA hydratase (EC 4.2.1.17)
- Curated sequence GFF2389: Enoyl-CoA hydratase [valine degradation] (EC 4.2.1.17)
- Ignore hits to items matching 4.2.1.17 when looking for 'other' hits
- Comment: Psest_2437 (GFF2389) is the enoyl-CoA hydrotase for both isoleucine and valine degradation, which implies that (S)-3-hydroxybutanoyl-CoA is a substrate. Q97MS7 is misannotated in BRENDA. BPHYT_RS17335 was misannotated as paaF; it is very similar to the ech H16_A3307, which is a different explanation for its role in phenylacetate utilization. Short-chain enoyl-CoA hydratases are sometimes given EC 4.2.1.17 instead, so those are ignored.
- Total: 8 characterized proteins
fadB: (S)-3-hydroxybutanoyl-CoA dehydrogenase
- Curated proteins or TIGRFams with EC 1.1.1.35
- Ignore hits to GFF1550 when looking for 'other' hits (Enoyl-CoA hydratase (EC 4.2.1.17))
- Comment: HP15_1512 (GFF1550) is annotated as enoyl-CoA hydratase but likely does this as well
- Total: 36 characterized proteins
dch: cyclohexa-1,5-diene-1-carboxyl-CoA hydratase
had: 6-hydroxycyclohex-1-ene-1-carbonyl-CoA dehydrogenase
oah: 6-oxocyclohex-1-ene-1-carbonyl-CoA hydratase
pimB: 3-oxopimeloyl-CoA:CoA acetyltransferase
Ch1CoA: cyclohex-1-ene-1-carbonyl-CoA dehydrogenase
badK: cyclohex-1-ene-1-carboxyl-CoA hydratase
badH: 2-hydroxy-cyclohexanecarboxyl-CoA dehydrogenase
badI: 2-ketocyclohexanecarboxyl-CoA hydrolase
pimD: pimeloyl-CoA dehydrogenase, large subunit
pimC: pimeloyl-CoA dehydrogenase, small subunit
pimF: 6-carboxyhex-2-enoyl-CoA hydratase
boxA: benzoyl-CoA epoxidase, subunit A
- Curated sequence Q9AIX6: Benzoyl-CoA oxygenase component A; Benzoyl-CoA 2,3-dioxygenase subunit A; Benzoyl-CoA dioxygenase reductase component; EC 1.14.13.208. benzoyl-CoA 2,3-epoxidase (subunit 2/2) (EC 1.14.13.208). benzoyl-CoA oxygenase component A subunit
- Comment: From EC 1.14.13.208
- Total: 1 characterized proteins
boxB: benzoyl-CoA epoxidase, subunit B
- Curated sequence Q9AIX7: Benzoyl-CoA oxygenase component B; Benzoyl-CoA 2,3-dioxygenase subunit B; Benzoyl-CoA dioxygenase oxygenase component; EC 1.14.13.208. benzoyl-CoA 2,3-epoxidase (subunit 1/2) (EC 1.14.13.208). benzoyl-CoA oxygenase component B (EC 1.14.13.208)
- Total: 1 characterized proteins
boxC: 2,3-epoxybenzoyl-CoA dihydrolase
boxD: 3,4-dehydroadipyl-CoA semialdehyde dehydrogenase
- Curated proteins or TIGRFams with EC 1.2.1.77
- Ignore hits to items matching 1.2.1.91 when looking for 'other' hits
- Comment: This reaction is similar to that of 3-oxo-5,6-dehydrosuberyl-CoA semialdehyde dehydrogenase (EC 1.2.1.91)
- Total: 2 characterized proteins
paaF: 2,3-dehydroadipyl-CoA hydratase
- Curated sequence P76082: enoyl-CoA hydratase (EC 4.2.1.17). 2,3-dehydroadipyl-CoA hydratase; Enoyl-CoA hydratase; EC 4.2.1.17
- Curated sequence MONOMER-15953: 2,3-dehydroadipyl-CoA hydratase (EC 4.2.1.17)
- Curated sequence BPHYT_RS17335: trans-2,3-dehydroadipyl-CoA hydratase (EC 4.2.1.17)
- UniProt sequence A0A2Z5MEB0: SubName: Full=Enoyl-CoA hydratase {ECO:0000313|EMBL:AXF15582.1};
- Comment: This reaction is associated with EC 4.2.1.17, which is very broad (enoyl-CoA hydratase). P76081 is E. coli paaF and MONOMER-15953 is the characterized enzyme from Pseudomonas sp. Y2. BPHYT_RS17335 from Burkholderia phytofirmans and H281DRAFT_05725 (A0A2Z5MEB0) from Paraburkholderia bryophila 376MFSha3.1 are required for phenylacetate utilization and are distantly related to E. coli paaF.
- Total: 4 characterized proteins
paaH: 3-hydroxyadipyl-CoA dehydrogenase
- Curated proteins or TIGRFams with EC 1.1.1.35
- Curated sequence GFF2749: 3-hydroxyacyl-CoA dehydrogenase PaaC (EC 1.1.1.-)
- Ignore hits to GFF1550 when looking for 'other' hits (Enoyl-CoA hydratase (EC 4.2.1.17))
- Comment: This step is described by 1.1.1.35, a broader term for 3-hydroxyacyl-CoA dehydrogenases. HP15_2693 (GFF2749) is involved in phenylalanine degradation via phenylacetyl-CoA and likely has this activity. HP15_1512 (GFF1550) is annotated as enoyl-CoA hydratase but likely has 3-hydroxyacyl-CoA dehydrogenase activity as well.
- Total: 37 characterized proteins
paaJ2: 3-oxoadipyl-CoA thiolase
- Curated proteins or TIGRFams with EC 2.3.1.174
- Ignore hits to items matching 2.3.1.223 when looking for 'other' hits
- UniProt sequence B2SYZ2: RecName: Full=3-oxoadipyl-CoA thiolase {ECO:0000256|ARBA:ARBA00012233}; EC=2.3.1.174 {ECO:0000256|ARBA:ARBA00012233};
- UniProt sequence A0A2Z5MFE9: RecName: Full=3-oxoadipyl-CoA thiolase {ECO:0000256|ARBA:ARBA00012233}; EC=2.3.1.174 {ECO:0000256|ARBA:ARBA00012233};
- UniProt sequence D8ITH5: RecName: Full=3-oxoadipyl-CoA thiolase {ECO:0000256|ARBA:ARBA00012233}; EC=2.3.1.174 {ECO:0000256|ARBA:ARBA00012233};
- Curated sequence GFF2751: Beta-ketoadipyl CoA thiolase (EC 2.3.1.-)
- Comment: Enzymes from B. phytofirmans and P. bryophila and H. seropedicae and M. adhaerens are included, as for paaJ1 above
- Total: 1 HMMs and 8 characterized proteins
pobA: 4-hydroxybenzoate 3-monooxygenase
ligA: protocatechuate 4,5-dioxygenase, alpha subunit
- Curated sequence MONOMER-15116: protocatechuate 4,5-dioxygenase alpha chain (EC 1.13.11.8). protocatechuate 4,5-dioxygenase (subunit 2/2) (EC 1.13.11.8)
- Curated sequence MONOMER-3165: protocatechuate 4,5-dioxygenase α subunit (EC 1.13.11.8; EC 1.13.11.57). protocatechuate 4,5-dioxygenase (EC 1.13.11.8)
- Ignore hits to items matching 1.13.11.8 when looking for 'other' hits
- Total: 2 characterized proteins
ligB: protocatechuate 4,5-dioxygenase, beta subunit
- Curated sequence G2IQQ3: protocatechuate 4,5-dioxygenase (subunit 1/2) (EC 1.13.11.8)
- Curated sequence MONOMER-15117: protocatechuate 4,5-dioxygenase beta chain (EC 1.13.11.8)
- Curated sequence MONOMER-3166: protocatechuate 4,5-dioxygenase β subunit (EC 1.13.11.8; EC 1.13.11.57)
- Ignore hits to items matching 1.13.11.8 when looking for 'other' hits
- Total: 3 characterized proteins
ligC: 2-hydroxy-4-carboxymuconate-6-semialdehyde dehydrogenase
ligI: 2-pyrone-4,6-dicarboxylate hydrolase
ligU: 4-oxalomesaconate tautomerase
- Curated proteins or TIGRFams with EC 5.3.2.8
- Curated sequence Q0KJL4: (4E)-oxalomesaconate Delta-isomerase; OMA isomerase; 1,3-allylic isomerase LigU; EC 5.3.3.-
- Comment: This isomerization can occur spontaneously, but the enzyme is probably required for good growth (i.e., fitness data for utilization of 4-vinylphenol or gallic acid). This enzyme is named gllD in MetaCyc. Note that ligU = Q0KJL4 is reported to carry out a different isomerization of 4-oxalomesaconate as part of this pathway. I believe this is just a question of whether to treat the product as the enol form (in UniProt) or the keto form (in MetaCyc).
- Total: 2 characterized proteins
ligJ: 4-carboxy-2-hydroxymuconate hydratase
- Curated proteins or TIGRFams with EC 4.2.1.83
- Curated sequence G2IQQ5: 2-keto-4-carboxy-3-hexenedioate hydratase; KCH hydratase; EC 4.2.1.-
- Comment: SwissProt G2IQQ5 = ligJ is described as using the enol form, (3Z)-2-oxo-4-carboxy-3-hexenedioate, of the substrate, but it yields the same product
- Total: 4 characterized proteins
ligK: 4-oxalocitramalate aldolase
praB: 2-hydroxymuconate 6-semialdehyde dehydrogenase
- Curated proteins or TIGRFams with EC 1.2.1.85
- Curated proteins or TIGRFams with EC 1.2.1.32
- Comment: EC 1.2.1.32 is 2-aminomuconate 6-semialdehyde dehydrogenase, but it is also reported to act on 2-hydroxymuconate-6-semialdehyde; many of these proteins are very similar to EC 1.2.1.85
- Total: 10 characterized proteins
praC: 2-hydroxymuconate tautomerase
praD: 2-oxohex-3-enedioate decarboxylase
xylF: 2-hydroxymuconate semialdehyde hydrolase
pcaH: protocatechuate 3,4-dioxygenase, alpha subunit
- Curated sequence A0A193DXA9: protocatechuate 3,4-dioxygenase (subunit 2/2) (EC 1.13.11.3)
- Curated sequence A8I4B7: protocatechuate 3,4-dioxygenase (EC 1.13.11.3)
- Curated sequence I0DHJ0: protocatechuate 3,4-dioxygenase (subunit 2/2) (EC 1.13.11.3)
- Curated sequence Q0SH26: protocatechuate 3,4-dioxygenase (subunit 1/2) (EC 1.13.11.3)
- Curated sequence CH_121294: protocatechuate 3,4-dioxygenase, alpha chain; EC 1.13.11.3
- Curated sequence MONOMER-14209: protocatechuate 3,4-dioxygenase type II α subunit (EC 1.13.11.3)
- Curated sequence MONOMER-3186: protocatechuate 3,4-dioxygenase α subunit (EC 1.13.11.3)
- Ignore hits to items matching 1.13.11.3 when looking for 'other' hits
- Total: 7 characterized proteins
pcaG: protocatechuate 3,4-dioxygenase, beta subunit
- Curated sequence A0A193DXP2: protocatechuate 3,4-dioxygenase (subunit 1/2) (EC 1.13.11.3)
- Curated sequence A8I4B3: protocatechuate 3,4-dioxygenase (EC 1.13.11.3)
- Curated sequence I0DHJ1: protocatechuate 3,4-dioxygenase (subunit 1/2) (EC 1.13.11.3)
- Curated sequence Q0SH27: protocatechuate 3,4-dioxygenase (subunit 2/2) (EC 1.13.11.3)
- Curated sequence CH_121290: protocatechuate 3,4-dioxygenase, beta chain; EC 1.13.11.3
- Curated sequence MONOMER-14210: protocatechuate 3,4-dioxygenase type II β subunit (EC 1.13.11.3)
- Curated sequence MONOMER-3185: protocatechuate 3,4-dioxygenase β subunit (EC 1.13.11.3)
- Ignore hits to items matching 1.13.11.3 when looking for 'other' hits
- Total: 7 characterized proteins
pcaB: 3-carboxymuconate cycloisomerase
- Curated proteins or TIGRFams with EC 5.5.1.2
- UniProt sequence A0A0C4YE08: SubName: Full=3-carboxy-cis,cis-muconate cycloisomerase {ECO:0000313|EMBL:AJG23937.1}; EC=5.5.1.2 {ECO:0000313|EMBL:AJG23937.1};
- Comment: RR42_RS32055 (A0A0C4YE08) is a somewhat diverged pcaB, confirmed by fitness data
- Total: 1 HMMs and 3 characterized proteins
pcaC: 4-carboxymuconolactone decarboxylase
pcaD: 3-oxoadipate enol-lactone hydrolase
praA: protocatechuate 2,3-dioxygenase
hcl: 4-hydroxybenzoyl-CoA ligase
- Curated proteins or TIGRFams with EC 6.2.1.27
- Curated sequence Q8GQN9: Benzoate--CoA ligase; Benzoyl-CoA synthetase; EC 6.2.1.25. benzoate-CoA ligase (EC 6.2.1.25). benzoate-CoA ligase (EC 6.2.1.32; EC 6.2.1.25)
- Ignore hits to items matching 6.2.1.25 when looking for 'other' hits
- Ignore hits to Q8VUF1 when looking for 'other' hits (benzoyl-CoA reductase (EC 1.3.7.8))
- Comment: benzoyl-CoA ligases (EC 6.2.1.25) may have this activity as well, i.e. see PMID:12897012 (discussing Q8GQN9). Q8VUF1 (probably a benzoyl-CoA ligase) is misannotated in BRENDA as benzoyl-CoA reductase.
- Total: 4 characterized proteins
hcrA: 4-hydroxybenzoyl-CoA reductase, alpha subunit
- Curated sequence O33819: 4-hydroxybenzoyl-CoA reductase subunit alpha; 4-HBCR subunit alpha; EC 1.3.7.9. 4-hydroxybenzoyl-CoA reductase α subunit (EC 1.3.7.9)
- Curated sequence MONOMER-14376: 4-hydroxybenzoyl-CoA reductase, α subunit (EC 1.3.7.9)
- Curated sequence MONOMER-17404: 4-hydroxybenzoyl-CoA reductase HbaC subunit (EC 1.3.7.9)
- Ignore hits to items matching 1.1.7.1 when looking for 'other' hits
- Ignore hits to items matching 1.3.7.9 when looking for 'other' hits
- Comment: EC 1.3.7.9 has been transferred to EC 1.1.7.1, but currently the annotations still use 1.3.7.9
- Total: 3 characterized proteins
hcrB: 4-hydroxybenzoyl-CoA reductase, beta subunit
- Curated sequence O33820: 4-hydroxybenzoyl-CoA reductase subunit beta; 4-HBCR subunit beta; EC 1.3.7.9. 4-hydroxybenzoyl-CoA reductase β subunit (EC 1.3.7.9)
- Curated sequence MONOMER-14377: 4-hydroxybenzoyl-CoA reductase, β subunit (EC 1.3.7.9)
- Curated sequence MONOMER-17405: 4-hydroxybenzoyl-CoA reductase HbaD subunit (EC 1.3.7.9)
- Ignore hits to items matching 1.1.7.1 when looking for 'other' hits
- Ignore hits to items matching 1.3.7.9 when looking for 'other' hits
- Total: 3 characterized proteins
hcrC: 4-hydroxybenzoyl-CoA reductase, gamma subunit
- Curated sequence O33818: 4-hydroxybenzoyl-CoA reductase subunit gamma; 4-HBCR subunit gamma; EC 1.3.7.9. 4-hydroxybenzoyl-CoA reductase γ subunit (EC 1.3.7.9)
- Curated sequence MONOMER-14378: 4-hydroxybenzoyl-CoA reductase, γ subunit (EC 1.3.7.9)
- Curated sequence MONOMER-17403: 4-hydroxybenzoyl-CoA reductase HbaB subunit (EC 1.3.7.9)
- Ignore hits to items matching 1.1.7.1 when looking for 'other' hits
- Ignore hits to items matching 1.3.7.9 when looking for 'other' hits
- Total: 3 characterized proteins
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About GapMind
Each pathway is defined by a set of rules based on individual steps or genes. Candidates for each step are identified by using
ublast (a fast alternative to protein BLAST)
against a database of manually-curated proteins (most of which are experimentally characterized) or by using
HMMer with enzyme models (usually from
TIGRFam). Ublast hits may be split across two different proteins.
A candidate for a step is "high confidence" if either:
- ublast finds a hit to a characterized protein at above 40% identity and 80% coverage, and bits >= other bits+10.
- (Hits to curated proteins without experimental data as to their function are never considered high confidence.)
- HMMer finds a hit with 80% coverage of the model, and either other identity < 40 or other coverage < 0.75.
where "other" refers to the best ublast hit to a sequence that is not annotated as performing this step (and is not "ignored").
Otherwise, a candidate is "medium confidence" if either:
- ublast finds a hit at above 40% identity and 70% coverage (ignoring otherBits).
- ublast finds a hit at above 30% identity and 80% coverage, and bits >= other bits.
- HMMer finds a hit (regardless of coverage or other bits).
Other blast hits with at least 50% coverage are "low confidence."
Steps with no high- or medium-confidence candidates may be considered "gaps."
For the typical bacterium that can make all 20 amino acids, there are 1-2 gaps in amino acid biosynthesis pathways.
For diverse bacteria and archaea that can utilize a carbon source, there is a complete
high-confidence catabolic pathway (including a transporter) just 38% of the time, and
there is a complete medium-confidence pathway 63% of the time.
Gaps may be due to:
- our ignorance of proteins' functions,
- omissions in the gene models,
- frame-shift errors in the genome sequence, or
- the organism lacks the pathway.
GapMind relies on the predicted proteins in the genome and does not search the six-frame translation. In most cases, you can search the six-frame translation by clicking on links to Curated BLAST for each step definition (in the per-step page).
For more information, see the paper from 2019 on GapMind for amino acid biosynthesis, the paper from 2022 on GapMind for carbon sources, or view the source code.
If you notice any errors or omissions in the step descriptions, or any questionable results, please let us know
by Morgan Price, Arkin group, Lawrence Berkeley National Laboratory