3-hydroxypropionate/4-hydroxybutyrate cycle

traitmech:000024 · CLASS · REVIEWED

An autotrophic carbon-fixation pathway that fixes two molecules of bicarbonate per turn via 3-hydroxypropionate and 4-hydroxybutyrate intermediates. It operates in aerobic and microaerophilic Crenarchaeota such as Sulfolobus and Metallosphaera.

Trait evidence (2)

  • DOI:10.1126/science.1149976

    Berg et al. described the 3-hydroxypropionate/4-hydroxybutyrate autotrophic CO2-assimilation pathway in Archaea (Sulfolobales).

  • DOI:10.1128/AEM.02473-10

    Berg review situates the 3HP/4HB cycle among the six recognized autotrophic carbon-fixation pathways.

3HP/4HB cycle fixes bicarbonate in aerobic Crenarchaeota

Evidence-backed causal sketch linking bicarbonate fixation via 3-hydroxypropionate and 4-hydroxybutyrate intermediates in Sulfolobales.

MECHANISTIC · The graph represents the experimentally characterized M. sedula 3HP/4HB cycle. Its bifunctional carboxylase is a three-component complex, so each component is recorded separately rather than treating one accession as the complete enzyme.

3HP/4HB cycle fixes bicarbonate in aerobic Crenarchaeota Interactive directed graph showing evidence-backed causal relationships for 3-hydroxypropionate/4-hydroxybutyrate cycle.

Edge evidence

  • hydrogencarbonate fixed by 3-hydroxypropionate/4-hydroxybutyrate cycle METPO:2007404

    Two molecules of bicarbonate are fixed per cycle turn.

  • 3-hydroxypropionate/4-hydroxybutyrate cycle contributes to carbon fixation RO:0002326

    The 3HP/4HB cycle is among the six recognized autotrophic pathways.

  • 3-hydroxypropionate/4-hydroxybutyrate cycle produces acetyl-CoA METPO:2007800

    A complete cycle turn fixes two bicarbonate to produce one acetyl-CoA.

  • 3-hydroxypropionate/4-hydroxybutyrate cycle forms intermediate succinyl-CoA

    Carboxylation of acetyl-CoA/propionyl-CoA yields succinyl-CoA from two bicarbonate.

  • succinyl-CoA is reduced to 4-hydroxybutyrate

    Succinyl-CoA is reduced to 4-hydroxybutyrate.

  • 4-hydroxybutyryl-CoA dehydratase dehydrates 4-hydroxybutyryl-CoA

    4-hydroxybutyryl-CoA is dehydrated to crotonyl-CoA.

  • 4-hydroxybutyryl-CoA dehydratase produces crotonyl-CoA METPO:2007800

    Dehydration of 4-hydroxybutyryl-CoA yields crotonyl-CoA.

  • crotonyl-CoA regenerates acetyl-CoA

    Crotonyl-CoA is processed/cleaved to regenerate acetyl-CoA for further cycle turns.

  • NADPH is required by 3-hydroxypropionate/4-hydroxybutyrate cycle

    Reductive steps of the cycle consume NADPH (about four NADPH per turn).

  • acetyl-CoA/propionyl-CoA carboxylase mediates carboxylation in 3-hydroxypropionate/4-hydroxybutyrate cycle

    Acetyl-CoA/propionyl-CoA carboxylase is the primary carboxylase fixing inorganic carbon.

Protein and taxon examples

Graph nodeProteinTaxonUniProt statusRole and evidence
acetyl-CoA/propionyl-CoA carboxylase UniProtKB:A4YD22
Biotin carboxylase (Msed_0147)
Metallosphaera sedula DSM 5348
NCBITaxon:399549
UNREVIEWED
retrieved 2026-08-24 · entry v93 · sequence v1

Alpha component (biotin carboxylase) of the three-component acetyl-CoA/propionyl-CoA carboxylase; this accession is not the complete enzyme.

  • DOI:10.1128/AEM.04146-13 Acetyl-CoA/propionyl-CoA carboxylase Msed_0147, Msed_0148, Msed_1375 The primary M. sedula pathway reconstruction maps all three loci to the bifunctional carboxylase. UniProtKB A4YD22 is the Msed_0147 component from the reference proteome.
acetyl-CoA/propionyl-CoA carboxylase UniProtKB:A4YD23
Biotin carboxyl carrier protein (Msed_0148)
Metallosphaera sedula DSM 5348
NCBITaxon:399549
UNREVIEWED
retrieved 2026-08-24 · entry v75 · sequence v1

Biotin-carboxyl-carrier component of the three-component acetyl-CoA/propionyl-CoA carboxylase; this accession is not the complete enzyme.

  • DOI:10.1128/AEM.04146-13 Acetyl-CoA/propionyl-CoA carboxylase Msed_0147, Msed_0148, Msed_1375 The primary M. sedula pathway reconstruction maps all three loci to the bifunctional carboxylase. UniProtKB A4YD23 is the Msed_0148 component from the reference proteome.
acetyl-CoA/propionyl-CoA carboxylase UniProtKB:A4YGI1
Propionyl-CoA carboxylase beta chain (Msed_1375)
Metallosphaera sedula DSM 5348
NCBITaxon:399549
UNREVIEWED
retrieved 2026-08-24 · entry v85 · sequence v1

Gamma carboxyltransferase component of the three-component acetyl-CoA/propionyl-CoA carboxylase; this accession is not the complete enzyme.

  • DOI:10.1128/AEM.04146-13 Acetyl-CoA/propionyl-CoA carboxylase Msed_0147, Msed_0148, Msed_1375 The primary M. sedula pathway reconstruction maps all three loci to the bifunctional carboxylase. UniProtKB A4YGI1 is the Msed_1375 component from the reference proteome.
4-hydroxybutyryl-CoA dehydratase UniProtKB:A4YGC7
Vinylacetyl-CoA Delta-isomerase (Msed_1321)
Metallosphaera sedula DSM 5348
NCBITaxon:399549
UNREVIEWED
retrieved 2026-08-24 · entry v82 · sequence v1

Msed_1321 protein biochemically assigned as 4-hydroxybutyryl-CoA dehydratase in the M. sedula pathway; the current UniProt label reflects alternate nomenclature.

  • DOI:10.1128/AEM.04146-13 4-hydroxybutyryl-CoA dehydratase (Msed_1321) Recombinant Msed_1321 was assigned to the dehydration step in the primary pathway reconstruction. UniProtKB A4YGC7 is the corresponding reference-proteome entry.

Provenance

Identifier source
TraitMech local identifier
Definition source
DOI:10.1126/science.1149976

Synonyms (1)

  • 3HP/4HB cycle RELATED_SYNONYM · DOI:10.1126/science.1149976

kg-microbe context

Matched 1 kg-microbe node via parent_proxy.

  • METPO:1000060 [-1.052, -1.766, -1.194, +0.291, …]

512-dim DeepWalkSkipGramEnsmallen embedding from kg-microbe (2026-04-25).

Nearest neighbors in embedding space

Top-8 cosine-similar METPO traits from the 2026-04-25 deepwalk (512-D).

Deep research

Generated by just research-trait; source: research/traits/metabolism/three_hydroxypropionate_four_hydroxybutyrate_cycle-deep-research-falcon.md

Unreviewed literature output — not curated TraitMech content Ontology identifiers suggested below have not been resolved against their ontologies, and some are known to be wrong. Check any CURIE against the source before using it.
# Curation report: 3-hydroxypropionate/4-hydroxybutyrate cycle

## Trait record

- **Trait label:** 3-hydroxypropionate/4-hydroxybutyrate cycle
- **Trait identifier:** `traitmech:000024`
- **Category / kind / status:** METABOLISM / CLASS / REVIEWED
- **Parent:** `traitmech:000019`
- **Synonym:** 3HP/4HB cycle

## 1. Scope and current definition

This trait should represent the **physiological capacity for autotrophic inorganic-carbon assimilation through the complete 3HP/4HB cycle**, rather than possession of an isolated enzyme or production of 3-hydroxypropionate. Its defining topology is:

**acetyl-CoA → malonyl-CoA → malonate semialdehyde → 3-hydroxypropionate → 3-hydroxypropionyl-CoA → acryloyl-CoA → propionyl-CoA → methylmalonyl-CoA → succinyl-CoA → succinate semialdehyde → 4-hydroxybutyrate → 4-hydroxybutyryl-CoA → crotonyl-CoA → (S)-3-hydroxybutyryl-CoA → acetoacetyl-CoA → two acetyl-CoA.**

Two bicarbonate/carbon-dioxide equivalents are incorporated as acetyl-CoA is converted to succinyl-CoA. The C4 intermediate is reduced through 4-hydroxybutyrate and ultimately cleaved into two acetyl-CoA molecules: one regenerates the carbon acceptor and the other is available for biosynthesis. The experimentally described *Metallosphaera sedula* cycle comprises 13 enzymes catalyzing 16 reactions. (hawkins2014conversionof4hydroxybutyrate pages 1-2, hawkins2013roleof4hydroxybutyratecoa pages 1-2, liu2021(s)3hydroxybutyrylcoadehydrogenasefrom pages 1-2)

### Validated biological scope

Two independently evolved variants should be represented under this trait:

1. **Crenarchaeal/Sulfolobales variant:** associated with thermoacidophilic, hydrogen-oxidizing, aerobic or microaerophilic organisms such as *Metallosphaera* and *Sulfolobus*.
2. **Thaumarchaeal/Nitrososphaeria variant:** associated with mesophilic, aerobic ammonia-oxidizing archaea such as *Nitrosopumilus maritimus*.

The variants are homologous in overall pathway topology but differ in several enzyme families and energetic costs. In particular, thaumarchaeal 3-hydroxypropionyl-CoA and 4-hydroxybutyryl-CoA synthetases are ADP-forming, whereas corresponding Sulfolobales enzymes are AMP-forming. This saves two ATP equivalents per cycle turn and is interpreted as adaptation to the low-energy, oligotrophic niche of ammonia-oxidizing archaea. (liu2021convergentevolutionof pages 1-2, johnson2024crystalstructureof pages 1-2)

### Boundary cases

- **Not the bacterial 3-hydroxypropionate bicycle.** That pathway shares acetyl-CoA carboxylation and 3HP chemistry but has a different cycle architecture and lacks the defining 4HB-to-two-acetyl-CoA regeneration arm.
- **Not the dicarboxylate/4-hydroxybutyrate cycle.** DC/4HB shares the succinyl-CoA→4HB→two-acetyl-CoA arm, but fixes carbon through ferredoxin-dependent pyruvate synthase and phosphoenolpyruvate carboxylase. The 3HP/4HB cycle instead uses acetyl-CoA/propionyl-CoA carboxylase twice and converts malonyl-CoA through 3HP and propionyl-CoA. DC/4HB is classically associated with anaerobic Thermoproteales and Desulfurococcales. (liu2021convergentevolutionof pages 1-2, liu2021(s)3hydroxybutyrylcoadehydrogenasefrom pages 1-2)
- **Partial 3HP production is insufficient.** Heterologous expression of the acetyl-CoA→3HP module does not establish the complete trait.
- **A single marker gene is insufficient.** Acetyl-CoA/propionyl-CoA carboxylases, enoyl-CoA hydratases, dehydrogenases, and thiolases have functions outside this cycle.
- **Genomic potential is not demonstrated physiology.** Bacterial MAGs predicted to encode the pathway and newly inferred geothermal carriers should remain provisional until pathway completeness, directionality, expression, and carbon-fixation flux are established. (garritano2022carbonfixationpathways pages 1-2, qi2024analysisofnearly pages 7-8)

## 2. Candidate nodes

### A. Pathway and process nodes

- `traitmech:000024` — 3-hydroxypropionate/4-hydroxybutyrate cycle
- `traitmech:000019` — supplied parent trait
- Autotrophic inorganic-carbon fixation — label-only unless the project already specifies its preferred GO/METPO term
- Bicarbonate assimilation
- Acetyl-CoA regeneration
- Biosynthetic acetyl-CoA production
- Crenarchaeal 3HP/4HB variant
- Thaumarchaeal modified 3HP/4HB variant

### B. Chemicals and cofactors

Recommended stable chemical identifiers, where unambiguous:

- Carbon dioxide — `CHEBI:16526`
- Hydrogencarbonate/bicarbonate — `CHEBI:17544`
- Acetyl-CoA — `CHEBI:15351`
- Malonyl-CoA — `CHEBI:15531`
- 3-hydroxypropionic acid/3-hydroxypropionate — `CHEBI:33404`
- Propionyl-CoA — `CHEBI:15539`
- Succinyl-CoA — `CHEBI:15380`
- 4-hydroxybutyrate — use a locally verified ChEBI record; do not assign from name alone because protonation-state records differ

Showing the first 60 of 278 lines of findings; the linked file also carries the run's front matter and the prompt it was given — read the full report.

Canonical examples (2)

Organisms cited as exemplars of this trait. Taxon ids are NCBITaxon and link out to the NCBI record.

  • Metallosphaera sedula NCBITaxon:43687 DOI:10.1126/science.1149976 Thermoacidophilic crenarchaeon where the 3-hydroxypropionate/4-hydroxybutyrate CO2-fixation cycle was characterized.
  • Metallosphaera sedula DSM 5348 NCBITaxon:399549 DOI:10.1128/AEM.04146-13 Reference strain used to map the three carboxylase components and Msed_1321 4-hydroxybutyryl-CoA dehydratase in biochemical reconstruction of the cycle.

Curation history

  1. · PROPOSED_FROM_RESEARCH · claude

    Proposed candidate METABOLISM trait (3-hydroxypropionate/4-hydroxybutyrate cycle); archaeal sub-variant of carbon fixation.

  2. · CURATED_CAUSAL_GRAPH · claude

    Added evidence-backed causal graph (3HP/4HB Sulfolobales bicarbonate fixation) with CHEBI/GO node groundings and RO/METPO predicate groundings; promoted PROPOSED to REVIEWED.

  3. · ENRICH_CAUSAL_GRAPH · claude

    Added 8 evidence-backed generic edges (8 new nodes) from the deep-research report.

  4. · GROUND_CAUSAL_PREDICATES · claude

    Grounded 2 causal-edge predicate_id field(s) via mappings/predicate_grounding.tsv (METPO:2000202×2).

  5. · GROUND_CAUSAL_NODES · claude

    Grounded 1 causal-node grounding field(s) via mappings/node_grounding.tsv (CHEBI:16474×1).

  6. · GROUND_CAUSAL_NODES · claude

    Grounded 2 causal-node grounding field(s) via mappings/node_grounding.tsv (CHEBI:16724×1, CHEBI:28522×1).

  7. · GROUND_CAUSAL_NODES · claude

    Grounded 2 causal-node grounding field(s) via mappings/node_grounding.tsv (UniProtKB:A0A650CEQ7×1, UniProtKB:A0A017H5F8×1).

  8. · RETRACT_DEAD_UNIPROT_GROUNDINGS · claude

    Retracted 1 UniProtKB grounding(s) whose accessions are deleted from UniProt; nodes demoted to label-only pending re-grounding (docs/GROUNDING_POLICY.md)

  9. · MIGRATE_MICROBE_DOMAIN_EDGES_PART2 · claude

    Re-grounded 2 causal edge(s) off microbe-domain METPO predicates onto their causal-graph counterparts (2 to produces), issue 301 part 2. The previous predicates are transitively rdfs:subPropertyOf METPO:2000001, whose rdfs:domain is METPO:1000525 (microbe), so a causal-graph subject entailed that the subject IS a microbe; CausalNodeTypeEnum has no organism member, so no such edge could ever satisfy the domain. Each replacement is a 1:1 mirror of its source predicate that changes only the domain, so the claim each edge makes is unchanged and directions are unchanged. The replacements are proposed in proposals/metpo_traitmech_v9 and are placeholder ids until METPO mints them.

  10. · REVIEW_UNIPROT_INSTANCE_GROUNDINGS · codex

    Reviewed 1 organism-specific UniProtKB grounding(s): replaced 0 with taxon-agnostic GO/InterPro terms and retracted 1 to label-only where no exact semantic term was supported (docs/GROUNDING_POLICY.md).

  11. · CURATE_PROTEIN_TAXON_EXAMPLE · codex

    Scoped the M. sedula 3HP/4HB mechanism, added all three DOI-backed carboxylase components plus Msed_1321 as taxon-paired UniProt examples, and documented why the complex and historical dehydratase labels remain reviewed label-only.