hglS

UniProt ID: Q88CC1
Organism: Pseudomonas putida KT2440
Review Status: COMPLETE
Aliases:
ydcJ PP_5260
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Gene Description

HglS is a 2-oxoadipate dioxygenase/decarboxylase that catalyzes the final step in D-lysine catabolism by converting 2-oxoadipate to D-2-hydroxyglutarate through an Fe(II)- and O2-dependent mechanism involving successive decarboxylation and intramolecular hydroxylation. The enzyme shows high specificity for 2-oxoadipate (KM ~0.01-0.06 mM) and is essential for growth on both L-lysine and D-lysine. HglS belongs to the DUF1338 family and represents the last missing enzymatic step in plant lysine catabolism that was identified through high-resolution crystal structures (PDB: 6W1G, 6W1H) which revealed the molecular basis for substrate specificity mediated by a conserved arginine residue (Arg74).

Proposed New Ontology Terms

2-oxoadipate dioxygenase activity

Definition: Catalysis of the oxidative decarboxylation of 2-oxoadipate to form D-2-hydroxyglutarate using Fe(II) and O2 as cofactors

Justification: HglS represents a specific type of dioxygenase activity with unique substrate specificity. A more specific term would better capture this distinct enzymatic function than the general dioxygenase activity term.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0016491 oxidoreductase activity
IEA
GO_REF:0000120
KEEP AS NON CORE
Summary: Accurate but general - HglS is specifically a dioxygenase/decarboxylase
Reason: While correct, this term is too general. The more specific dioxygenase activity term better captures HglS function.
GO:0051213 dioxygenase activity
IEA
GO_REF:0000043
ACCEPT
Summary: Correct and specific - HglS performs dioxygenase activity on 2-oxoadipate
Reason: Accurately describes the core molecular function. HglS is a Fe(II)-dependent dioxygenase that incorporates O2 during the oxidative decarboxylation of 2-oxoadipate.
GO:0019477 L-lysine catabolic process
IMP
PMID:31064836
Massively parallel fitness profiling reveals multiple novel ...
NEW
Summary: Core biological process - HglS is essential for lysine catabolism. Term updated from obsolete GO:0006554 to GO:0019477.
Reason: HglS catalyzes a key step in lysine breakdown and deletion mutants cannot grow on lysine. Original term GO:0006554 was obsoleted in favor of more specific terms.
Supporting Evidence:
PMID:31064836
Massively Parallel Fitness Profiling Reveals Multiple Novel Enzymes in Pseudomonas putida Lysine Metabolism.
GO:0042180 ketone metabolic process
IDA
PMID:31064836
Massively parallel fitness profiling reveals multiple novel ...
NEW
Summary: Specific metabolic process - HglS processes 2-oxoadipate (a ketone derivative)
Reason: The substrate 2-oxoadipate is a ketone derivative and the reaction involves ketone metabolism.
Supporting Evidence:
PMID:31064836
Massively Parallel Fitness Profiling Reveals Multiple Novel Enzymes in Pseudomonas putida Lysine Metabolism.
GO:0006520 amino acid metabolic process
IMP
PMID:31064836
Massively parallel fitness profiling reveals multiple novel ...
NEW
Summary: Broad metabolic process - HglS functions in amino acid catabolism
Reason: As part of lysine catabolism, HglS participates in amino acid metabolic processes.
Supporting Evidence:
PMID:31064836
Massively Parallel Fitness Profiling Reveals Multiple Novel Enzymes in Pseudomonas putida Lysine Metabolism.

Core Functions

Fe(II)-dependent dioxygenase that catalyzes oxidative decarboxylation of 2-oxoadipate to D-2-hydroxyglutarate with high substrate specificity (KM ~0.01-0.06 mM)

Supporting Evidence:
  • PMID:31064836
    involves the direct conversion of 2OA to d-2HG
  • PMID:32523014
    HglS exhibited activity only with 2OA, and displayed no detectable and statistically significant activity above background

Essential metabolic enzyme for bacterial growth on both L-lysine and D-lysine as carbon sources

Supporting Evidence:
  • PMID:31064836
    A Ξ”PP_5260 strain was unable to grow on either isomer of lysine
  • PMID:31064836
    PP_5260, showed a significant fitness defect...near other d-lysine catabolic genes in the P. putida genome

References

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Suggested Questions for Experts

Q: How does HglS coordinate with other enzymes in the lysine catabolic pathway?

Suggested experts: Bacterial metabolism specialists, Enzyme biochemists, Systems biologists

Q: What is the evolutionary origin of the DUF1338 family and how did substrate specificity evolve?

Suggested experts: Evolutionary biochemists, Comparative genomics researchers, Structural biologists

Q: Can HglS be engineered for biotechnological production of D-2-hydroxyglutarate or related compounds?

Suggested experts: Enzyme engineers, Synthetic biologists, Metabolic engineers

Suggested Experiments

Experiment: Systematic comparison of HglS substrate specificity across different 2-oxo acids to fully define the substrate scope and identify any secondary activities.

Type: Comparative enzymatic analysis

Experiment: Site-directed mutagenesis of residues beyond Arg74 to understand the complete molecular basis for substrate specificity and catalytic mechanism.

Type: Structural-functional analysis

Experiment: Comprehensive analysis of DUF1338 family members across bacteria and eukaryotes to understand evolutionary conservation and functional divergence.

Type: Phylogenetic analysis

Experiment: Complete mapping of lysine catabolism pathway in P. putida to understand HglS integration with upstream and downstream enzymes.

Type: Metabolic pathway analysis

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