Acetate kinase (ACK) , CAS No.9027-42-3

CAS: 9027-42-3 Cat. No.: A1522969 EC Number: 232-818-0

GRADE & PURITY Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™, ≥94%(SDS-PAGE), ≥10 U/mg enzyme powder; ≥100 U/mg protein

Accession #

P0A6A3

Bioactivity

≥10 U/mg enzyme powder; ≥100 U/mg protein

Overview

Acetate kinase (ACK) is an enzyme widely distributed in bacteria and archaea. In the presence of ATP and divalent cations, it catalyzes the phosphorylation of acetate and promotes the production of acetyl-CoA. Often used in biochemical research.

Specifications

Product Name

Acetate kinase (ACK) , CAS No.9027-42-3

Synonyms

ATP:Acetate phosphotransferase | acetate kinase (phosphorylating) | acetic kinase | acetokinase | AK | ackA | ECK2290

Grade

ActiBioPure™, Bioactive, High Performance, Recombinant, EnzymoPure™

Specifications & Purity

Bioactive,Recombinant,ActiBioPure™,High Performance,EnzymoPure™,≥94%(SDS-PAGE),≥10 U/mg enzyme powder; ≥100 U/mg protein

Bioactivity

≥10 U/mg enzyme powder; ≥100 U/mg protein

Accession #

P0A6A3

CAS

9027-42-3

Enzyme Commission Number

EC 2.7.2.1

Molecule Type

Enzyme

Product Line

Biological Products

Storage and Shipping

Concentration

≥10 U/mg enzyme powder; ≥100 U/mg protein

Reconstitution

Reconstitute in water to a concentration of 10mg/ml.

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at 2-8℃ short term (12 months). Store at -20℃ long term (24 months). Upon reconstitution, it is recommended to aliquot. Avoid freeze/thaw cycle.

Unit definition

Under conditions of pH 7.6 and 25°C, one unit can phosphorylate 1.0 μmole of acetic acid into acetyl phosphate per minute.

Application Protocols

Below are example, literature-derived protocols to help design experiments with acetate kinase. They are general guidance and not item-specific specifications.

A) Continuous ACK activity assay (PK/LDH-coupled):

• Prepare buffer: 50 mM HEPES, pH 7.6; 10 mM MgCl2; 100 mM KCl.
• Add components: 2 mM ATP; 50 mM sodium acetate; 0.2 mM NADH; 2 mM phosphoenolpyruvate; excess pyruvate kinase and lactate dehydrogenase (per kit instructions).
• Equilibrate at 25–30°C; initiate reaction with ACK. Monitor A340 decrease; calculate rates using ε340(NADH)=6.22 mM−1·cm−1.

B) ATP regeneration module (reverse direction):

• Assemble: 50 mM Tris-HCl, pH 7.8; 10 mM MgCl2; 100 mM KCl; 10–20 mM acetyl phosphate; ADP at the expected steady-state burden (e.g., 0.1–1 mM); partner ATP-dependent enzyme(s).
• Start reaction with ACK; adjust enzyme loading to maintain target ATP level. Refresh acetyl phosphate as consumed.

C) Colorimetric acetyl phosphate detection (hydroxamate):

• React sample with neutral hydroxylamine to form acetyl hydroxamate, then develop color with Fe3+ under acidic conditions; measure A540–550 nm (literature). Include acetate blanks to correct background.

General notes:

• Keep Mg2+ constant and avoid EDTA in assay mixes.
• Validate linearity with respect to enzyme concentration and time.
• For low-abundance ACK, add BSA (0.1 mg/mL) to minimize adsorption.

Exact enzyme units, protein concentration, and buffer composition for this SKU: Not specified; refer to CoA/Spec Sheet.

Biological Roles

Acetate kinase (EC 2.7.2.1) is a central enzyme in bacterial acetate metabolism and energy homeostasis (literature).

• Metabolic context: In the Pta–Ack pathway, phosphotransacetylase (PTA) and ACK interconvert acetyl-CoA, acetyl phosphate, and acetate, coupling acetyl-CoA metabolism to ATP/ADP balance. The ACK reaction direction depends on cellular energy charge and metabolite concentrations.
• Energy economy: ACK enables ATP synthesis from acetyl phosphate and ADP under fermentative conditions, contributing to substrate-level phosphorylation in many bacteria.
• Regulatory/signaling aspects: Acetyl phosphate serves as a small-molecule phosphodonor to some two-component response regulators, linking carbon flux to transcriptional responses (literature). Thus, ACK activity influences pools of acetyl phosphate that can affect regulatory networks.
• Physiological diversity: ACK homologs occur across bacteria and archaea, with mesophilic and thermophilic variants tailored to native niches. Some organisms carry multiple ack genes with differential expression.
• Structural biology: Many ACKs are homodimers with conserved ATP- and acetate-binding motifs; Mg2+ is essential for catalysis. Mutational studies identify residues coordinating phosphoryl transfer and metal binding, illuminating catalytic mechanism.

Note: These roles describe general biology of acetate kinase and are provided for research context only. They do not specify the source organism or sequence of this catalog item (not provided).

Buffer Applications

This product is an enzyme rather than a buffer component; however, appropriate buffer selection is critical for its activity and stability.

General/literature guidance for ACK assay and storage buffers (not item-specific):

• Storage buffer (example): 20–50 mM Tris-HCl, pH 7.4–8.0; 50–200 mM NaCl; 1–5 mM DTT or TCEP; 10–50% glycerol for frozen storage. Optional 0.1–0.5 mM EDTA to protect against adventitious metals (omit in assays requiring Mg2+). Exact composition for this item is not specified; refer to CoA/Spec Sheet.
• Assay buffer (example): 50 mM HEPES or Tris, pH 7.5–8.0; 5–10 mM MgCl2; substrate pair at desired concentrations (e.g., 1–10 mM ATP with 10–50 mM acetate, or 5–20 mM acetyl phosphate with 0.1–2 mM ADP for ATP regeneration). Include 100–150 mM NaCl if needed for stability.
• Additives: 0.1 mg/mL BSA can reduce adsorption losses in dilute assays. Avoid high phosphate if it interferes with detection or equilibria.
• Temperature: Perform assays at 25–37°C for mesophilic ACKs unless a thermostable enzyme is used.

These conditions are typical literature starting points. Optimize for your specific substrates, detection methods, and desired direction of the ACK reaction. Always check the product’s datasheet for any required cofactors or stabilizers supplied with the enzyme.

Green Alternatives

Enzymatic phosphate-transfer systems are generally greener than purely chemical phosphorylation due to aqueous operation, mild conditions, and high selectivity. Within enzymatic ATP-regeneration options, acetate kinase can be compared to creatine kinase (CK) and polyphosphate kinase (PPK).

Comparison (literature, illustrative):

• Acetate kinase (ACK)
  • Phosphate donor: Acetyl phosphate (readily prepared and water-soluble).
  • Byproducts: Acetate/acetyl phosphate interconversion; minimal auxiliary salts.
  • Pros: Simple components; avoids creatine derivatives; works well in phosphate-buffered systems at controlled concentrations.
  • Cons: Acetyl phosphate can nonenzymatically acetylate nucleophiles at high concentrations; stability requires fresh prep or cold storage.
• Creatine kinase (CK)
  • Donor: Phosphocreatine.
  • Pros: Widely used in molecular biology kits; robust.
  • Cons: Introduces creatine/phosphate salts; cost and solubility considerations.
• Polyphosphate kinase (PPK)
  • Donor: Inorganic polyphosphate.
  • Pros: Very economical phosphate source; scalable.
  • Cons: PolyP heterogeneity; potential interactions with metal ions and nucleic acids.

Greener practice tips:

• Run reactions in water-based buffers; avoid organic cosolvents when possible.
• Recycle cofactors via enzymatic regeneration to minimize ATP consumption and waste.
• Use minimal effective Mg2+ and maintain closed systems to limit phosphate precipitation.

Note: Selection should consider downstream compatibility and propensity of acetyl phosphate to acetylate sensitive biomolecules at elevated concentrations (mitigated by buffering and temperature control).

Pharmaceutical Uses

This product is supplied for research use only. It is not a drug, diagnostic, or therapeutic. There are, however, research and manufacturing-adjacent contexts where acetate kinase can be useful as a processing aid or tool enzyme (literature/general):

• Bioprocess and cell-free systems: ACK can support ATP regeneration using acetyl phosphate in cell-free protein synthesis or enzymatic production lines, reducing exogenous ATP consumption and improving sustainability of in vitro pathways.
• Analytical QC: Coupled enzymatic assays involving ACK may be used in process analytics to quantify acetate, acetyl phosphate, or ATP/ADP ratios in development settings.
• Enabling chemistry: As part of enzyme cascades, ACK facilitates mild, aqueous phosphorylation steps, which can be valuable in green chemistry approaches to produce intermediates for research compounds.
• Excipients/formulation: Not applicable—ACK is an active enzyme reagent, not a formulation excipient.

Regulatory/compendial status: Not specified for this item; no pharmacopeial monograph is implied.

If planning to use acetate kinase as a processing aid in manufacturing research, align acceptance criteria (activity units, purity, host-cell protein/DNA, endotoxin) with internal quality systems. For any clinical manufacturing, consult appropriate regulatory guidance; this catalog item is not supplied for clinical use.

Physical Properties

Proteins do not have classical small-molecule physical constants such as BP/MP; functional properties are governed by conformation, buffer, and temperature.

Item-specific specifications:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Extinction coefficient, concentration, additives (e.g., glycerol, salts): Not specified for this item; refer to CoA/Spec Sheet.

General/literature guidance for acetate kinase (for context only; not item specifications):

• Typical quaternary state: Homodimer (literature), though some ACKs behave as dimers/tetramers depending on ionic strength.
• Optimal pH (activity): Often near neutral to mildly basic, pH ~6.5–8.0, depending on organism and assay conditions (literature).
• Temperature profile: Many mesophilic ACKs show robust activity at 25–37°C; thermophilic variants are active at higher temperatures (literature).
• Essential cofactors: Mg2+ (or Mn2+ in some cases) required for ATP binding/turnover (literature).
• Substrate solubility: Acetate and acetyl phosphate are water-soluble; ATP/ADP used as Mg2+ complexes in aqueous buffer (literature).

Refractive index, pKa, logP, vapor pressure, and related small-molecule parameters: Not applicable to a protein enzyme.

For precise formulation, concentration, and stabilizers of this catalog item, consult the product’s CoA/Spec Sheet.

Quality & Grades

This item is designated as Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™. While exact acceptance criteria are not specified for this SKU, the following describes typical implications of these designations for protein reagents.

• Bioactive: Supplied in a form that retains catalytic function under recommended assay conditions. Activity is expected, but specific units/assay parameters are not specified for this item; refer to CoA/Spec Sheet.
• Recombinant: Produced in a heterologous host to enhance consistency, supply, and purity. The expression host, tag, and construct details are not specified for this item.
• ActiBioPure™ (brand designation): Typically indicates stringent control of activity preservation, low contaminant burden (e.g., nucleases/proteases), and suitable buffer composition to maintain function. Exact specifications are not provided here.
• High Performance: Generally denotes high specific activity and low lot-to-lot variability—important for coupled assays and ATP-regeneration systems. Item-specific metrics are not provided.
• EnzymoPure™ (brand designation): Emphasizes high protein purity appropriate for sensitive biochemical workflows (e.g., cell-free systems). Absorbance ratios, SDS-PAGE profiles, and host-cell protein/DNA limits are not specified for this item.

What to verify on receipt (recommended):

• Review CoA for purity (SDS-PAGE), specific activity assay format, storage buffer/additives, and endotoxin/host contaminants if relevant.
• Confirm absence of interfering activities (e.g., ATPases, phosphatases) if using in coupled assays. Obtain inhibitor screen if critical for your workflow.

Note: Always match the supplier’s assay definition to your intended application to ensure comparable performance.

Reaction & Applications

Functional role (literature): Acetate kinase catalyzes acetate + ATP ⇌ acetyl phosphate + ADP. The reaction is readily coupled for driving biochemical pathways and analytical assays.

Key application areas:

• ATP regeneration: Use ACK with acetyl phosphate to convert ADP back to ATP in cell-free protein synthesis, kinase assays, and ligase/polymerase workflows. Compared to creatine kinase systems, ACK avoids creatine/phosphocreatine and produces acetate as a benign byproduct (when run in the reverse direction with acetyl phosphate).
• Acetyl phosphate generation: Forward reaction from acetate + ATP provides acetyl phosphate for two-component system studies, nonenzymatic phosphorylation reactions, or as a high-energy intermediate for metabolic pathway driving (literature).
• Metabolic engineering: In the Pta–Ack pathway, ACK can be employed in vitro to balance ATP/acetyl-CoA flux, or in semi-synthetic cascades with phosphotransacetylase (PTA) to interconvert acetyl-CoA and acetyl phosphate.
• Analytical/coupled assays: Monitor ACK activity by coupling ADP production to pyruvate kinase/lactate dehydrogenase (PK/LDH) with NADH consumption at 340 nm; or detect acetyl phosphate via hydroxamate/Fe(III) colorimetric assays (literature).

Practical tips:

• Maintain Mg2+ in the reaction (e.g., 5–10 mM) and control pH near neutrality to slightly basic.
• Remove phosphate or chelators that may sequester Mg2+ or interfere with detection chemistries.
• Pre-equilibrate enzyme on ice; initiate reactions by substrate addition to minimize lag.
• For ATP-regeneration, titrate acetyl phosphate and enzyme to match ADP load and turnover demand; validate steady-state ATP levels empirically.

Item-specific activity units/assay definitions: Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Typical literature conditions for acetate kinase reactions (provide a starting framework—optimize empirically for your system):

• Temperature: 25–37°C for mesophilic enzymes; adjust based on source enzyme’s thermal profile.
• pH: 7.0–8.0 (Tris-HCl, HEPES, or MOPS buffers at 25°C). Activity often peaks near mildly basic pH.
• Mg2+: 5–10 mM MgCl2 required for ATP/ADP binding and catalysis; higher levels may be inhibitory in some systems.
• Substrate concentrations:
  • Forward (acetate + ATP → acetyl phosphate + ADP): ATP 1–10 mM; acetate 10–200 mM.
  • Reverse/ATP regeneration (acetyl phosphate + ADP → acetate + ATP): acetyl phosphate 5–50 mM; ADP 0.1–2 mM, scaled to ATP demand.
• Ionic strength: 50–200 mM NaCl can stabilize dimeric state; adjust to compatibility of partner enzymes.
• Coupled assay example (PK/LDH, literature):
  1. Buffer: 50 mM Tris-HCl, pH 7.8; 10 mM MgCl2; 100 mM KCl.
  2. Add ATP and acetate (or ADP and acetyl phosphate for reverse).
  3. Include pyruvate kinase + phosphoenolpyruvate (for ADP recycling) and lactate dehydrogenase + NADH; monitor A340 decrease as a readout of ADP formation.
• Reaction times: Minutes to hours depending on enzyme loading and target conversion.
• Atmosphere: Ambient; no special exclusion of oxygen required.

Kinetic constants (Km/kcat) vary widely by homolog and are not specified for this item. Always confirm linear ranges of activity for your chosen conditions.

Safety & Handling

• GHS classification, signal word, pictograms, and H-statements: Not specified for this item; consult the SDS for authoritative safety information.
• General hazard profile: Enzyme protein for laboratory use. Not intended for human or animal use, diagnostic, or therapeutic applications. Avoid inhalation of powders/aerosols and contact with skin/eyes.
• PPE: Laboratory coat, gloves (nitrile), and safety glasses as a minimum. Use a biological safety cabinet if handling lyophilized powder to minimize aerosolization.
• Handling: Work on ice or at 2–8°C during prolonged manipulations to preserve bioactivity. Avoid repeated freeze–thaw cycles (explicit product instruction). Aliquot immediately upon receipt if multiple uses are anticipated.
• Storage incompatibilities: Avoid strong oxidizers, proteases, and extremes of pH or temperature that may denature proteins. Maintain chelators (e.g., EDTA) out of assay mixes if Mg2+ is required for activity.
• First-aid overview: If skin/eye contact occurs, rinse with plenty of water for at least 15 minutes; remove contaminated clothing. If inhaled, move to fresh air. If ingested, rinse mouth. Seek medical attention according to institutional guidelines. Defer to SDS.
• Spills: Absorb small spills with inert material; clean surfaces with aqueous detergent. For lyophilized material, gently wet to suppress dust before cleanup.
• Waste: Dispose of according to institutional biosafety and chemical waste procedures.
• Research use note: For research use only (per product data).

Solvent Selection

As a protein enzyme, acetate kinase is handled in aqueous buffers rather than organic solvents.

General buffer considerations (literature; adapt to your system):

• Buffer systems: Tris-HCl, HEPES, or MOPS at pH ~7.0–8.0 commonly support activity. Avoid phosphate at high concentrations in assays that monitor acetyl phosphate/ATP equilibria, unless validated.
• Ionic strength: 50–200 mM NaCl often stabilizes quaternary structure while minimizing nonspecific interactions. Optimize to balance stability and activity.
• Metal ions: Mg2+ (5–20 mM) is typically required for catalysis; avoid excess EDTA in reaction mixes. Trace Mn2+ can modulate kinetics in some homologs.
• Stabilizers: 10–50% glycerol (v/v) for frozen storage; 0.1–1 mg/mL BSA can reduce adsorption losses in dilute working solutions. Presence/amounts for this SKU are not specified.
• Reducing environment: 1–5 mM DTT or TCEP can maintain thiol integrity if required; confirm compatibility with downstream analytics.

When to choose alternatives:

• High-temperature processes: Consider thermophilic ACK variants if working >45°C (literature).
• ATP-regeneration in phosphate-sensitive systems: Polyphosphate kinase (PPK) or creatine kinase (CK) may be preferable depending on metabolite compatibility (see Green Alternatives).

Non-aqueous/organic solvents are generally incompatible with enzyme activity; if cosolvents are necessary for substrates, keep below denaturation thresholds (e.g., <5–10% v/v for many alcohols, literature) and revalidate activity.

Storage & Reconstitution

• Storage conditions (product data): Store at −20°C. Avoid repeated freezing and thawing.
• Shipping (product data): Shipped in an ice chest with ice pads to maintain cold chain.

Recommended good practices (general; not item-specific):

• Upon receipt: Immediately place at −20°C (or as indicated on the label). If frequent use is expected, prepare small working aliquots to prevent freeze–thaw cycles.
• Aliquoting: Use low-protein-binding tubes. If the formulation permits, include 10–50% glycerol for storage at −20°C to maintain a liquid state and prevent ice crystallization; verify compatibility with the supplied buffer.
• Reconstitution: If supplied lyophilized, reconstitute with cold, oxygen-free water or assay/storage buffer (e.g., 20–50 mM Tris-HCl, pH 7.5–8.0, 50–200 mM NaCl, 1–5 mM DTT). Gently invert; avoid vortexing/foaming. If supplied as a solution, mix gently before aliquoting. The exact form of this item (solution vs lyophilized) is not specified; refer to CoA/Spec Sheet.
• Working solutions: Keep on ice during use. For long incubations, include stabilizers (e.g., BSA 0.1 mg/mL) to reduce adsorption losses, if compatible with your assay.
• Freeze–thaw guidance: One-time thaw on ice; dispense aliquots; promptly refreeze unused material at −20°C or below. Avoid more than one freeze–thaw.
• Light/oxidation: Protect from strong light and air exposure. Consider adding reducing agent (e.g., 1–5 mM DTT/TCEP) if compatible.

Any item-specific concentration, stabilizers, or shelf-life: Not specified for this item; refer to CoA/Spec Sheet.

Structure & Identity

Acetate kinase (ACK) is a phosphoryl-transfer enzyme that catalyzes the reversible conversion: acetate + ATP ⇌ acetyl phosphate + ADP.

• SKU: A1522969
• Product name: Acetate kinase (ACK)
• CAS: 9027-42-3
• EC number: 2.7.2.1 (literature)
• Synonyms (literature): Ack, acetokinase, acetate kinase A (ackA).
• Macromolecule class: Protein enzyme (recombinant; Bioactive) — precise source organism and construct not specified for this item.
• Molecular formula: Not defined for proteins; item-specific composition not specified.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. Note: many bacterial ACK monomers are ~40–45 kDa (literature), but this varies by source and tag.
• SMILES / InChI / InChIKey: Not applicable for proteins; not provided for this item.
• Structural features (literature): Typical ACKs are homodimers with each monomer comprising an N- and C-terminal domain forming an active-site cleft that binds ATP/Mg2+ and acetate/acetyl phosphate. A highly conserved P-loop/Walker A-like phosphate-binding region and residues coordinating Mg2+ are common. No stereocenters in the polymer sense; local chiral centers in amino acids.
• 2D/functional description: Polypeptide chain forming a dimeric enzyme; active site mediates transfer of the terminal phosphate of ATP to acetate to form acetyl phosphate, or the reverse under suitable conditions.

Item-specific identity details (sequence, tag, host, isoform): Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

In chemoenzymatic synthesis and biocatalysis, acetate kinase provides a versatile phosphoryl-transfer step under mild, aqueous conditions (literature):

• ATP/ADP cycling: ACK can regenerate ATP from ADP using acetyl phosphate, enabling ATP-dependent ligations, kinases, and synthetases to operate with lower ATP input. This reduces costs and waste in preparative biocatalytic cascades.
• High-energy intermediate access: Generation of acetyl phosphate in situ allows driving unfavorable equilibria or feeding downstream enzymes such as phosphotransacetylase (PTA) to form acetyl-CoA, linking to acylation or CoA-dependent steps.
• Thermodynamic leverage: By coupling ACK to sinks for products (e.g., trapping acetyl phosphate via PTA/CoA or consuming ADP via downstream reactions), complex pathways can be pushed toward desired products.
• Orthogonality: Compared to creatine kinase or polyphosphate kinase systems, ACK uses small, inexpensive donors/acceptors (acetate/acetyl phosphate) and can be integrated with phosphate-tolerant downstream enzymes.
• Analytical synthetic control: Activity can be monitored continuously via PK/LDH NADH assays at 340 nm, allowing tight process feedback.

Integration tips:

• Balance Mg2+ and ionic strength for maximal stability without inhibiting partner enzymes.
• Control acetyl phosphate concentration to minimize nonenzymatic acetylation of amines/hydroxyls; operate at moderate temperature and buffered pH.
• Validate compatibility with thiol-containing cofactors and protein tags in multi-enzyme cascades.

Target Specificity

This section is typically applicable to affinity reagents such as antibodies or ligands. For acetate kinase, a catalytic enzyme, “target” refers to its substrates and cofactors rather than antigen specificity.

• Substrate specificity (literature): Catalyzes reversible phosphoryl transfer between acetate/acetyl phosphate and ATP/ADP. Some homologs may accept propionate at reduced efficiency; broad acid tolerance is limited and homolog-dependent.
• Cofactor requirements: Mg2+ is essential; Mn2+ can substitute in some systems with altered kinetics (literature).

No antibody clone, epitope, isotype, or species reactivity applies to this product. Item-specific substrate scope or engineered changes in specificity (if any) are not specified; refer to CoA/Spec Sheet if provided.