Recombinant Actinobacillus pleuropneumoniae serotype 3 Galactokinase (galK)

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In Stock
Cat. No.
BT55615
Source
Yeast
Synonyms
galK; APJL_1012Galactokinase; EC 2.7.1.6; Galactose kinase
Purity
>85% (SDS-PAGE)
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Cat. No.
BT55615
Source
E.coli
Synonyms
galK; APJL_1012Galactokinase; EC 2.7.1.6; Galactose kinase
Purity
>85% (SDS-PAGE)
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Cat. No.
BT55615
Source
E.coli
Synonyms
galK; APJL_1012Galactokinase; EC 2.7.1.6; Galactose kinase
Purity
>85% (SDS-PAGE)
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Cat. No.
BT55615
Source
Baculovirus
Synonyms
galK; APJL_1012Galactokinase; EC 2.7.1.6; Galactose kinase
Purity
>85% (SDS-PAGE)
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Cat. No.
BT55615
Source
Mammalian cell
Synonyms
galK; APJL_1012Galactokinase; EC 2.7.1.6; Galactose kinase
Purity
>85% (SDS-PAGE)
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Description

Definition and Basic Characterization

Recombinant Actinobacillus pleuropneumoniae serotype 3 galactokinase (galK) is an enzymatically active protein produced through genetic engineering. It is derived from the galK gene of A. pleuropneumoniae, a Gram-negative pathogen responsible for porcine pleuropneumonia. This recombinant protein is expressed in heterologous systems such as Escherichia coli, yeast, baculovirus, or mammalian cells, ensuring high purity (≥85% by SDS-PAGE) and functionality . Galactokinase catalyzes the ATP-dependent phosphorylation of galactose to galactose-1-phosphate, a critical step in the Leloir pathway of galactose metabolism.

Research Applications

Recombinant galK is utilized in multiple research contexts:

  1. Enzyme Kinetics: Studies on substrate specificity, catalytic efficiency, and inhibitor screening .

  2. Pathogen Metabolism: Investigating carbon source utilization in A. pleuropneumoniae .

  3. Diagnostic Development: As an antigen for serological assays targeting A. pleuropneumoniae infections .

  4. Structural Biology: Crystallography to resolve galactokinase’s 3D structure for drug design .

Key Steps in Recombinant Production

ParameterDetails
Expression VectorpGEX-KG (GST-tagged) or similar systems
InductionIsopropyl-β-D-thiogalactoside (IPTG) at 1 mM
Purification MethodAffinity chromatography (e.g., glutathione-Sepharose)
Storage ConditionsLyophilized powder at -20°C/-80°C in Tris/PBS buffer with 6% trehalose

Quality Control

  • Purity validated via SDS-PAGE .

  • Activity confirmed using spectrophotometric assays measuring ATP consumption or galactose-1-phosphate production .

Challenges and Future Directions

  1. Solubility Issues: Recombinant galK may form inclusion bodies in E. coli, requiring optimized expression conditions .

  2. Functional Redundancy: Other sugar kinases in A. pleuropneumoniae might compensate for galK knockout, complicating phenotypic studies .

  3. Therapeutic Potential: galK is not yet explored as a vaccine candidate, unlike other A. pleuropneumoniae antigens (e.g., LPS components) .

Key Research Findings

  • Genetic Conservation: The galK gene is conserved across A. pleuropneumoniae serotypes, suggesting its metabolic indispensability .

  • Regulatory Networks: galK expression may be modulated by global regulators like ArcA, which represses competing pathways under anaerobic conditions .

  • Biotechnological Utility: Recombinant galK serves as a tool for synthesizing galactose-1-phosphate in vitro, aiding glycobiology research .

Product Specs

Form
Lyophilized powder. We will ship the in-stock format preferentially. If you have special format requirements, please note them when ordering.
Lead Time
Delivery time varies by purchase method and location. Consult your local distributor for specifics. All proteins ship with standard blue ice packs. Request dry ice in advance (extra fees apply).
Notes
Avoid repeated freeze-thaw cycles. Store working aliquots at 4°C for up to one week.
Reconstitution
Briefly centrifuge the vial before opening. Reconstitute in sterile deionized water to 0.1-1.0 mg/mL. Add 5-50% glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final glycerol concentration is 50%.
Shelf Life
Shelf life depends on storage conditions, buffer components, temperature, and protein stability. Liquid form: 6 months at -20°C/-80°C. Lyophilized form: 12 months at -20°C/-80°C.
Storage Condition
Store at -20°C/-80°C upon arrival. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag type is determined during manufacturing. If you require a specific tag, please inform us and we will prioritize its development.
Synonyms
galK; APJL_1012Galactokinase; EC 2.7.1.6; Galactose kinase
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-384
Protein Length
full length protein
Purity
>85% (SDS-PAGE)
Species
Actinobacillus pleuropneumoniae serotype 3 (strain JL03)
Target Names
galK
Target Protein Sequence
MKPQQLATQK FSEHYGYSAA QTVFAPGRVN IIGEHTDYND GFVMPCAINY GMAVSFSKRD DSVWRVYAID IDEQDEFDLS RPIEPSEHKW ANYVRGVVKY IQEKCPEFKQ GADLAMTSDV PMSSGLSSSA ALEISIGKTA QVLGDLPLSL AEIALIGQQA ENKFVGANCG NMDQLTSALG QKDQVIMIDC RSLEITPTPV PHGYSIAIIN SNVKHDLVTG EYNSRRQECE FAAKFFGVKA LRDVTPAQFI ERAAELQAEN ELAYKRAKHI ISENQRVLEA VEALQAKDMV KLGQLMAGSH DSMRDDFEIT IPEIDYLVEL AQVAIGKNGG ARMTGGGFGG CIVCLVPDEK VEHLRRIIAD NYEKQTGIKE TFHLCTACDG VHLI
Uniprot No.
B0BPT5

Target Background

Function
Catalyzes the transfer of ATP's gamma-phosphate to D-galactose, forming alpha-D-galactose-1-phosphate (Gal-1-P).
Database Links

KEGG: apj:APJL_1012

Protein Families
GHMP kinase family, GalK subfamily
Subcellular Location
Cytoplasm.

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