Recombinant Escherichia coli 6-phosphofructokinase (pfkA)

Shipped with Ice Packs
In Stock
Cat. No.
BT36066
Source
Yeast
Synonyms
pfkA; EC55989_4394ATP-dependent 6-phosphofructokinase isozyme 1; ATP-PFK 1; Phosphofructokinase 1; EC 2.7.1.11; 6-phosphofructokinase isozyme I; Phosphohexokinase 1
Purity
>85% (SDS-PAGE)
Quick Inquiry
Cat. No.
BT36066
Source
E.coli
Synonyms
pfkA; EC55989_4394ATP-dependent 6-phosphofructokinase isozyme 1; ATP-PFK 1; Phosphofructokinase 1; EC 2.7.1.11; 6-phosphofructokinase isozyme I; Phosphohexokinase 1
Purity
>85% (SDS-PAGE)
Quick Inquiry
Cat. No.
BT36066
Source
E.coli
Synonyms
pfkA; EC55989_4394ATP-dependent 6-phosphofructokinase isozyme 1; ATP-PFK 1; Phosphofructokinase 1; EC 2.7.1.11; 6-phosphofructokinase isozyme I; Phosphohexokinase 1
Purity
>85% (SDS-PAGE)
Quick Inquiry
Cat. No.
BT36066
Source
Baculovirus
Synonyms
pfkA; EC55989_4394ATP-dependent 6-phosphofructokinase isozyme 1; ATP-PFK 1; Phosphofructokinase 1; EC 2.7.1.11; 6-phosphofructokinase isozyme I; Phosphohexokinase 1
Purity
>85% (SDS-PAGE)
Quick Inquiry
Cat. No.
BT36066
Source
Mammalian cell
Synonyms
pfkA; EC55989_4394ATP-dependent 6-phosphofructokinase isozyme 1; ATP-PFK 1; Phosphofructokinase 1; EC 2.7.1.11; 6-phosphofructokinase isozyme I; Phosphohexokinase 1
Purity
>85% (SDS-PAGE)
Quick Inquiry

Description

Biochemical and Genetic Characteristics

Catalytic Function:
PfkA is an allosteric enzyme that regulates glycolytic flux through its sensitivity to metabolic effectors like phosphoenolpyruvate (PEP, inhibitor) and ADP (activator) . Unlike its isozyme PfkB (encoded by pfkB), PfkA accounts for 80–95% of total phosphofructokinase activity in E. coli under standard conditions .

Gene Structure and Expression:

  • The pfkA gene is located at 76 minutes on the E. coli genetic map .

  • Recombinant PfkA has been synthesized in vitro using plasmid systems (e.g., pLC 16-4), achieving ~30% of the in vivo synthesis efficiency .

Kinetic Parameters:

Substrate/CofactorValue (Native)Value (Recombinant)ConditionsSource
Fructose-6-phosphateS0.5=0.7 mMS_{0.5} = 0.7 \, \text{mM}Km=0.25 mMK_m = 0.25 \, \text{mM}pH 7, 75°C
ATPKm=0.2 mMK_m = 0.2 \, \text{mM}Km=0.15 mMK_m = 0.15 \, \text{mM}pH 8.2, RT
V<sub>max</sub> (forward)360 U/mg320 U/mg75°C

2.1. Metabolic Engineering Applications

  • Dynamic Knockdown: A pfkA knockdown strain (IB1643) with tunable expression showed near-wild-type growth but redirected carbon flux toward non-glycolytic pathways, enabling enhanced production of compounds like inositol .

  • Co-utilization of Carbon Sources: Deletion of pfkA and zwf (glucose-6-phosphate dehydrogenase) in E. coli shifted metabolism from the Embden-Meyerhof-Parnas (EMP) pathway to the Entner-Doudoroff (ED) pathway, improving galactose utilization .

Phenotypic Effects of PfkA Manipulation

Mutation TypeGrowth on GlucoseFitness In Vivo (UTI Model)Key ObservationsSource
ΔpfkAImpairedReducedReliance on gluconeogenesis for fructose-6-phosphate synthesis
ΔpfkA ΔpfkBNo growthCompetitive in bladderSuppressed latent enzymes; enhanced survival in amino acid-rich environments
pfkA knockdown (IPTG-inducible)Tunable growthN/ARedirected flux to non-glycolytic pathways

Industrial and Biotechnological Relevance

  • Biosynthetic Pathways: Disruption of pfkA in triple mutants (ΔpfkA ΔpfkB Δzwf) enabled novel pathways for dihydroxyacetone (DHA) production via fructose-6-phosphate aldolase (FsaA) .

  • Antimicrobial Targeting: PfkA suppression in uropathogenic E. coli (UPEC) reduced virulence by altering ATP levels and lipopolysaccharide structure .

Challenges and Future Directions

  • Enzyme Overexpression: High PfkA activity can lead to metabolic imbalance, necessitating precise regulation in engineered strains .

  • Thermodynamic Constraints: The reversibility of the PfkA reaction under certain conditions complicates flux control in synthetic pathways .

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 times vary 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, storage 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 receipt. 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
pfkA; EC55989_4394ATP-dependent 6-phosphofructokinase isozyme 1; ATP-PFK 1; Phosphofructokinase 1; EC 2.7.1.11; 6-phosphofructokinase isozyme I; Phosphohexokinase 1
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-320
Protein Length
full length protein
Purity
>85% (SDS-PAGE)
Species
Escherichia coli (strain 55989 / EAEC)
Target Names
pfkA
Target Protein Sequence
MIKKIGVLTS GGDAPGMNAA IRGVVRSALT EGLEVMGIYD GYLGLYEDRM VQLDRYSVSD MINRGGTFLG SARFPEFRDE NIRAVAIENL KKRGIDALVV IGGDGSYMGA MRLTEMGFPC IGLPGTIDND IKGTDYTIGF FTALSTVVEA IDRLRDTSSS HQRISVVEVM GRYCGDLTLA AAIAGGCEFV VVPEVEFSRE DLVNEIKAGI AKGKKHAIVA ITEHMCDVDE LAHFIEKETG RETRATVLGH IQRGGSPVPY DRILASRMGA YAIDLLLAGY GGRCVGIQNE QLVHHDIIDA IENMKRPFKG DWLDCAKKLY
Uniprot No.
B7LA13

Target Background

Function
Catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate by ATP, the first committed step of glycolysis.
Database Links

KEGG: eck:EC55989_4394

Protein Families
Phosphofructokinase type A (PFKA) family, ATP-dependent PFK group I subfamily, Prokaryotic clade "B1" sub-subfamily
Subcellular Location
Cytoplasm.

Quick Inquiry

Personal Email Detected
Please use an institutional or corporate email address for inquiries. Personal email accounts ( such as Gmail, Yahoo, and Outlook) are not accepted. *

Category

Service

THE BIOTEK
THE BioTek is a global biotech company offering highly purified proteins for research in cancer, apoptosis, development, endocrinology, immunology, neuroscience, proteases, and stem cells.
  • Contact
  • Address: 1925 E Maple Ave, El Segundo, CA 90245 United States
  • Phone : +1 201-285-7826
  • Email : info@thebiotek.com
  • Hours : Mon.-Fri. 9:00 AM - 5:00 PM
© Copyright 2025 TheBiotek. All Rights Reserved.