pykF E.Coli

Pyruvate Kinase I E.Coli Recombinant

Cat. No.

BT14499

Source

E.coli.

Synonyms

Pyruvate kinase I, PK-1, pykF, b1676, JW1666.

Appearance

Sterile Filtered colorless solution.

Purity

Greater than 95% as determined by SDS-PAGE.

Usage

THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

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Description

pykF E.Coli Recombinant produced in E.Coli is a single, non-glycosylated polypeptide chain containing 494 amino acids (1-470 a.a) and having a molecular mass of 53.3kDa.
pykF is fused to a 24 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

Product Specs

Introduction

Pyruvate kinase I (pykF) is an enzyme that plays a crucial role in glycolysis. As a member of the pyruvate kinase family, pykF catalyzes the final step of glycolysis: the conversion of phosphoenolpyruvate (PEP) and ADP to pyruvate and ATP. This process requires a magnesium ion and is essential for energy production in cells.

Description

Recombinant pykF from E. coli is produced as a single, non-glycosylated polypeptide chain. This protein consists of 494 amino acids (with amino acids 1-470 being part of the pykF sequence) and has a molecular weight of 53.3 kDa. For purification and detection purposes, a 24 amino acid His-tag is fused to the N-terminus of the protein. The purification process utilizes proprietary chromatographic techniques.

Physical Appearance

Clear, colorless solution that has been sterilized by filtration.

Formulation

The pykF protein is supplied in a solution at a concentration of 1 mg/ml. The solution is buffered with 20mM Tris-HCl at a pH of 8.0 and contains 10% glycerol for stability.

Stability

For short-term storage (up to four weeks), the protein solution can be stored at 4°C. For extended storage, it is recommended to freeze the solution at -20°C. Adding a carrier protein such as HSA or BSA (0.1%) is advised for long-term storage. To maintain protein integrity, avoid repeated freeze-thaw cycles.

Purity

The purity of the pykF protein is greater than 95%, as determined by SDS-PAGE analysis.

Synonyms

Pyruvate kinase I, PK-1, pykF, b1676, JW1666.

Source

E.coli.

Amino Acid Sequence

MGSSHHHHHH SSGLVPRGSH MGSHMKKTKI VCTIGPKTES EEMLAKMLDA GMNVMRLNFS HGDYAEHGQR IQNLRNVMSK TGKTAAILLD TKGPEIRTMK LEGGNDVSLK AGQTFTFTTD KSVIGNSEMV AVTYEGFTTD LSVGNTVLVD DGLIGMEVTA IEGNKVICKV LNNGDLGENK GVNLPGVSIA LPALAEKDKQ DLIFGCEQGV DFVAASFIRK RSDVIEIREH LKAHGGENIH IISKIENQEG LNNFDEILEA SDGIMVARGD LGVEIPVEEV IFAQKMMIEK CIRARKVVIT ATQMLDSMIK NPRPTRAEAG DVANAILDGT DAVMLSGESA KGKYPLEAVS IMATICERTD RVMNSRLEFN NDNRKLRITE AVCRGAVETA EKLDAPLIVV ATQGGKSARA VRKYFPDATI LALTTNEKTA HQLVLSKGVV PQLVKEITST DDFYRLGKEL ALQSGLAHKG DVVVMVSGAL VPSGTTNTAS VHVL.

Product Science Overview

Pyruvate Kinase Isoforms

In Escherichia coli (E. coli), there are two isoforms of pyruvate kinase: PykF (pyruvate kinase I) and PykA (pyruvate kinase II). PykF is the more predominant form and is allosterically regulated by fructose-1,6-bisphosphate (FBP). This regulation ensures that the enzyme’s activity is tightly controlled in response to the cell’s metabolic needs.

Recombinant Expression in E. Coli

Recombinant expression of pyruvate kinase I in E. coli involves cloning the gene encoding the enzyme into an expression vector, which is then introduced into E. coli cells. The bacteria are cultured under conditions that induce the expression of the recombinant protein. This method allows for the production of large quantities of the enzyme, which can be purified and used for various biochemical studies.

Biochemical Properties

Pyruvate kinase I from E. coli has a molecular weight of approximately 58 kDa. The enzyme is typically supplied as a lyophilized powder and is stable at -20°C. It is often used in research to study enzyme kinetics, metabolic regulation, and the effects of various inhibitors on glycolysis.

Applications

Recombinant pyruvate kinase I is widely used in biochemical research. It serves as a model to study the regulation of glycolysis and the role of allosteric effectors in enzyme activity. Additionally, it is used in the development of assays to screen for potential inhibitors that could be used as antibiotics or in the treatment of metabolic disorders.

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pykF E.Coli

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