The pyruvate synthase β-subunit (PorB) is a key component of multi-subunit pyruvate:ferredoxin oxidoreductase (PFOR), an enzyme complex central to anaerobic metabolism in many archaea and bacteria. Unlike monomeric oxidative decarboxylases, these multi-polypeptide PFORs separate functional domains into distinct subunits, with PorB corresponding to “domain VI” in the canonical six-domain architecture (NCBI COG Pathway – Pyruvate oxidation).

COG Classification: PorB is part of COG1013, clustering with β-subunits of 2-oxoacid:ferredoxin oxidoreductases (NCBI COG PorB entry).

Genome Example: Thermococcus nautili PorB is annotated under GeneID 82170787 (NCBI Gene record).

Operon Organization: In Hydrogenobacter thermophilus, PorB occurs in the porEDABG gene cluster, tightly linked with other PFOR subunits (PubMed: porEDABG study).

Domain Mapping: In Methanosarcina acetivorans, PorB forms domain VI of PFOR, hosting the proximal [4Fe-4S] cluster close to the TPP active site (PMC Structural Mapping).

Cofactor Binding: PorB’s Fe–S cluster relays electrons from the TPP-bound intermediate to the ferredoxin-binding δ-subunit.

Subunit Interfaces: β–δ contacts stabilize the electron transfer path, critical for catalytic efficiency.

PFOR catalyzes:

1. Oxidative Decarboxylation: Pyruvate → Acetyl-CoA + CO₂ (electrons to ferredoxin)
2. Reductive Carboxylation (Pyruvate Synthase mode): Acetyl-CoA + CO₂ + reduced ferredoxin → Pyruvate

Mechanistic details of TPP-dependent decarboxylation are reviewed in PubMed: TPP enzyme mechanism and PMC Review.

UC Berkeley ggKbase contig with PorB

Archival PorB contig in archaea

RefSeq M. jannaschii genome

1. Anaerobic Purification: Maintain Fe–S cluster integrity with strict anoxic conditions.
2. Cofactor Saturation: Ensure full TPP binding before assays (Gonzaga University TPP notes).
3. Electron Carrier Matching: Use ferredoxin or flavodoxin depending on the organism (PMC H. pylori POR study).

PorB’s proximal Fe–S cluster placement and electron tunneling interfaces make it a control point for carbon flux in anaerobes. Engineering PorB can modulate CO₂ fixation efficiency, making it a target for:

1. Biohydrogen production
2. CO utilization pathways
3. Synthetic autotrophic circuits

The β-subunit PorB is more than just a component of pyruvate synthase, it’s a redox and structural hub that integrates TPP chemistry, Fe–S electron transfer, and multi-subunit assembly into a single functional module. For metabolic engineers, structural biologists, and enzymologists, PorB offers both a fascinating subject for study and a practical lever for pathway optimization. Shop here.

1. What is Pyruvate Synthase Subunit PorB?

PorB is the β-subunit of the pyruvate:ferredoxin oxidoreductase (PFOR) complex, an enzyme responsible for interconverting pyruvate and acetyl-CoA under anaerobic conditions. In multi-subunit PFORs, PorB corresponds to domain VI in the six-domain architecture, hosting the proximal [4Fe-4S] cluster that transfers electrons from the thiamine diphosphate (TPP) active site to the ferredoxin-binding δ-subunit (NCBI COG1013 entry).

2. How does PorB contribute to pyruvate metabolism?

PorB plays a key role in electron transfer during both:

1. Oxidative decarboxylation: Pyruvate → Acetyl-CoA + CO₂
2. Reductive carboxylation: Acetyl-CoA + CO₂ + reduced ferredoxin → Pyruvate

This electron flow depends on the integrity of its Fe–S cluster and inter-subunit contacts (PMC: PFOR structural analysis).

3. In which organisms is PorB found?

PorB homologs occur in:

1. Anaerobic archaea such as Methanosarcina acetivorans (NCBI Genome)
2. Anaerobic bacteria including Hydrogenobacter thermophilus (PubMed)
3. Pathogens such as Helicobacter pylori, where PorB participates in pyruvate:flavodoxin oxidoreductase (PMC Study)

4. What cofactors does PorB use?

PorB contains a [4Fe-4S] iron–sulfur cluster, coordinated by conserved cysteine residues. This cluster relays electrons from the TPP-bound intermediate to downstream electron carriers (PMC Review on TPP enzymes).

5. How can I identify PorB in genome data?

Researchers use:

1. KEGG K-number K00170 in pathway searches
2. COG1013 in COG functional profiles (NCBI COG Pathway)
3. BLASTp searches using curated β-subunit sequences from model organisms (NCBI BLAST)

6. How is PorB studied experimentally?

1. Anaerobic purification to protect Fe–S clusters
2. Spectrophotometric assays with benzyl viologen as an electron sink
3. Mutagenesis to probe Fe–S coordination sites

7. What is the difference between PorB and PorA?

1. PorA (α-subunit): Contains the TPP-binding site and catalytic residues for pyruvate decarboxylation
2. PorB (β-subunit): Houses the proximal Fe–S cluster and mediates electron transfer
3. Both work together in the multi-subunit enzyme complex (PMC Structural Mapping).

8. Is PorB related to PorB in Neisseria species?

No. Neisseria PorB is an outer membrane porin, completely unrelated in function and structure to PFOR β-subunit PorB. The name overlap is coincidental.

9. Can PorB be engineered for biotechnology?

Yes. modifying PorB’s Fe–S environment can alter:

1. Redox potential
2. Electron transfer rate
3. Substrate channeling efficiency
4. This has implications for CO₂ fixation, biohydrogen production, and synthetic metabolism (DOE JGI IMG terms).

10. Where can I find structural data for PorB?

Crystal and cryo-EM structures for PorB-containing complexes are available in:

1. Protein Data Bank (PDB) entries via RCSB PDB
2. Literature: Structural data in PMC Open Access

11. How does PorB fit into the reductive TCA cycle?

In autotrophic archaea and bacteria, PorB-containing pyruvate synthase converts CO₂ + acetyl-CoA → pyruvate, feeding into gluconeogenesis and amino acid biosynthesis (NCBI Pathway Map).

12. How conserved is PorB across species?

PorB is highly conserved in anaerobes, with strong sequence conservation in Fe–S ligating cysteine motifs, supporting its ancient role in energy metabolism (PMC Comparative OFOR analysis).