Penicillinase, CAS No.9001-74-5

CAS: 9001-74-5 Cat. No.: P1508160 Número EC: 232-628-8

GRADE & PURITY Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™, Sterile, expressed in E.coli;≥10 M units/mL

Accession #

P00809

Expression system

E. coli

Bioactivity

≥10 M units/mL

Descripción general

Penicillinase, also known as type I β-lactamase (EC 3.5.2.6), can hydrolyze the β-lactam ring of β-lactam antibiotics, thereby abrogating their antimicrobial activity. This product is a recombinant penicillinase produced by genetic engineering technology. A high-expression engineering strain of penicillinase is constructed, followed by fermentation and purification to obtain a product with high purity and high activity. The product features strong specificity. It can effectively hydrolyze β-lactam antibiotics, including benzylpenicillin (penicillin G), ampicillin, amoxicillin, pivampicillin, carbenicillin, ticarcillin, furbenicillin, mezlocillin, etc.

Scope of Application

It can be used for the sterility test and microbial limit test of β-lactam antibiotic raw materials and their preparations, namely the sterility test for β-lactam antibiotics specified in Pharmacopoeia of the People's Republic of China (2020 Edition, Volume 4, Appendix: Sterility Test) and the microbial limit test specified in the corresponding appendix. It is also used as a reagent for the sterility test specified in USP-NF General Chapters 71 STERILITY TEST.

Usage Instructions

1. Membrane Filtration Method

① Dilute 1 ml of penicillinase to 1000 ml with sterile water to prepare the washing solution. Sterilize all the equipment to be used by an appropriate method and soak them in the washing solution for 10 minutes.

② Take the prescribed amount of the test sample, filter it according to the specified method, and rinse the filter membrane with an appropriate volume of the washing solution prepared in step ①.

③ Separately dilute 1 ml of penicillinase to 200 ml with sterile water. Add 10 ml of this solution to each culture medium, and store the remaining solution at 2–8 °C for later use.

④ Inoculate the filter membrane into the culture medium according to the specified method.

2. Direct Inoculation Method

Take the prescribed amount of the test sample (in accordance with the pharmacopoeia), mix it well, add an appropriate volume of sterile penicillinase solution (in accordance with the pharmacopoeia), and inoculate the mixture into each tube of culture medium. Alternatively, directly inoculate the test sample into the culture medium containing sterile penicillinase.

Specifications

Product Name

Penicillinase, CAS No.9001-74-5

Sinónimos

β-lactamase type I | β-Lactamase I

Grado

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

Especificaciones y pureza

Bioactive,Recombinant,ActiBioPure™,High Performance,EnzymoPure™,sterile,expressed in E.coli;≥10 M units/mL

Mecanismos bioquímicos y fisiológicos

This protein is a beta-lactamase with a substrate specificity for penicillins. The class A beta-lactamase family has a specific amino-acid numbering system, sometimes called Ambler or ABL numbering and often misspelt as Amber. A multiple sequence alignme

Bioactividad

≥10 M units/mL

Número de adhesión

P00809

CAS

9001-74-5

Número de la Comisión de Enzimas

EC 3.5.2.6

Tipo de molécula

Enzyme

Product Line

biological_products

Almacenamiento y envío

Concentración

expressed in E.coli;≥10 M units/mL

Condiciones de almacenamiento de almacenamiento

Store at 2-8°C

Enviado en

Wet ice

Estabilidad y almacenamiento

Store at 2-8℃ long term (24 months). Upon receipt, it is recommended to aliquot.

Definición de unidad

One activity unit is defined as the amount of enzyme required to hydrolyze 1 unit of penicillin G in 1 hour under conditions of pH 7.0 and 37˚C.

Application Protocols

Tested applications, recommended dilutions, and positive controls: Not specified for this item; refer to CoA/Spec Sheet.

Note: As general background only (not product-specific), penicillin hydrolysis assays commonly monitor ΔA240 nm for penicillin G in phosphate buffer at 25–37°C. For this specific catalog item, follow the CoA/Spec Sheet for activity units, assay buffer, and any validated procedures.

Biological Roles

Penicillinase (β-lactamase; EC 3.5.2.6) is a bacterial enzyme that confers resistance to β-lactam antibiotics by hydrolyzing the β-lactam amide bond.

• Natural occurrence (literature):
  • Widely distributed in Gram-positive and Gram-negative bacteria; secreted or periplasmic depending on species.
  • Multiple molecular classes (A–D) with distinct active-site chemistry (serine- vs. metallo-β-lactamases) and substrate spectrums; “penicillinase” typically refers to penicillin-preferring serine β-lactamases (class A).
• Mechanistic biology:
  • Active-site serine executes nucleophilic attack to form an acyl-enzyme intermediate; a general base activates water for deacylation and product release.
  • Substrate recognition involves interactions with the β-lactam ring and side chain; mutations modulate spectrum and inhibitor susceptibility (literature).
• Research significance:
  • Model enzyme for studying antibiotic resistance mechanisms, inhibitor design, and fitness costs of resistance.
  • Useful positive control in assays validating β-lactamase inhibitor activity and β-lactam stability testing.
• No clinical or diagnostic claims: This product is RUO. Any discussion refers to fundamental biochemistry and microbiology.

Note: Sequence, cellular localization, and regulation are organism- and variant-specific; consult sequence records and primary literature when a particular β-lactamase lineage is required for experimental design.

Buffer Applications

This enzyme is typically handled and used in neutral, aqueous buffers optimized for activity and stability.

• Common buffer systems (general, literature guidance):
  • Phosphate buffer (50 mM), pH 7.0–7.5; ionic strength 100–150 mM NaCl for stability.
  • HEPES or MOPS (25–50 mM), pH 7.2–7.6; minimal temperature-dependent pKa drift.
  • Tris-HCl (25–50 mM), pH 7.5–8.0; note temperature-dependent pH shift (−0.028 pH/°C approx.).
• Additives (as compatible with your assay):
  • 5–10% glycerol for cold storage stability; 0.1 mg/mL BSA to limit surface adsorption.
  • Low concentrations of non-ionic detergents (e.g., 0.01% Tween-20) can reduce sticking; verify assay compatibility.
  • Avoid EDTA only if using metallo-β-lactamases; for serine β-lactamases, moderate EDTA typically has little effect, but confirm experimentally (literature).
• Example preparation (illustrative, literature):
  • 50 mM sodium phosphate, 150 mM NaCl, pH 7.2; chill to 2–8°C. Add enzyme to achieve desired activity units/mL. Incubate with penicillin substrate at 25–37°C; monitor by UV at 240 nm or HPLC.
• Sterile workflows: Filter buffers (0.22 µm) and handle aseptically to maintain the product’s Sterile grade.

Note: Exact buffer composition and compatible excipients for this item are not specified; consult the CoA/Spec Sheet for formulation details.

Green Alternatives

Using an enzyme catalyst for β-lactam ring opening is itself a greener alternative to harsh chemical hydrolysis.

Comparison of β-lactam hydrolysis approaches (general, literature):

• Enzymatic (Penicillinase):
  • Pros: Aqueous media, near-neutral pH, ambient temperature; high selectivity; minimal byproducts; easy workup.
  • Cons: Protein stability constraints; potential cost; sensitive to denaturants; requires removal/inactivation if incompatible downstream.
• Acid/base chemical hydrolysis:
  • Pros: Low-cost reagents; no biocatalyst handling.
  • Cons: Strong acids/bases, elevated temperatures; non-selective degradation; increased waste neutralization; potential sample damage.
• Oxidative methods:
  • Pros: Can be rapid.
  • Cons: Non-specific oxidation; hazardous oxidants; poor compatibility with biomolecules.

Greener practice recommendations:

• Use buffered aqueous solutions at pH ~7–8 and ambient temperature when feasible.
• Employ minimal effective enzyme quantity and recover/inactivate enzyme to reduce residual biological load.
• Choose recyclable, low-leach labware; minimize single-use plastics.
• For scale-up, consider immobilized penicillinase on reusable supports to improve stability, enable continuous flow, and simplify separation.

Overall, enzymatic hydrolysis aligns with green chemistry principles (energy efficiency, safer solvents, reduced waste) relative to purely chemical alternatives.

Pharmaceutical Uses

Research/manufacturing context only—no therapeutic claims.

• Typical roles in pharmaceutical R&D and manufacturing (general, literature):
  • Process aid to degrade trace β-lactam antibiotics in research materials or intermediates when interference with analytical assays is a concern.
  • Analytical reagent in QC/QA for β-lactam content determination via enzymatic conversion to penicilloic acids with subsequent detection (UV/HPLC).
  • Tool enzyme in inhibitor screening campaigns targeting β-lactamases.
• Formulation considerations:
  • If used in process development experiments, enzyme is typically applied in aqueous buffers at neutral pH and removed/inactivated afterward (e.g., filtration, heat, or pH shift) to avoid carryover.
  • Sterile grade facilitates aseptic manipulations in clean laboratory environments.
• Regulatory note:
  • This product is RUO (research use only). Not manufactured under cGMP for drug substance/product; not intended as an excipient or API.

Item-specific pharmacopeial status, activity units, and acceptance criteria: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific physicochemical specifications have not been provided for this recombinant enzyme preparation.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• State: Protein reagent (aqueous sterile preparation or concentrate; exact form not specified).
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Literature: common β-lactamases ~28–35 kDa depending on source and tag.
• Isoelectric point (pI): Not specified for this item; refer to CoA/Spec Sheet.
  • Literature: enzyme pI varies by sequence; many class A β-lactamases pI ~5–7 (literature, sequence-dependent).
• Solubility: Highly soluble in physiological aqueous buffers (general for proteins). Specific solubility not specified for this item; refer to CoA/Spec Sheet.
• Extinction coefficient, A280-to-concentration: Not specified for this item; refer to CoA/Spec Sheet (sequence-dependent).
• Stability window (general, literature): Active in neutral to slightly alkaline buffers (e.g., pH ~6.5–8.5), activity typically retained at 2–8°C for short-term; thermal denaturation at elevated temperatures.
• BP/MP, density, logP, RI: Not applicable to proteins; not specified for this item; refer to CoA/Spec Sheet.

Note: For precise activity units, buffer composition, and protein concentration, consult the product’s CoA/Spec Sheet; these are sequence- and formulation-dependent and not provided here.

Quality and Grades

This product is supplied as a recombinant, sterile enzyme for research use with enhanced purity/performance branding.

• Grade/Purity (as listed): Bioactive, Recombinant, ActiBioPure™, High Performance, EnzymoPure™, Sterile.

  • Bioactive: Supplied in a form that retains enzymatic activity suitable for biochemical assays (item-specific activity not specified; refer to CoA/Spec Sheet).
  • Recombinant: Produced via heterologous expression to support batch-to-batch consistency and traceability.
  • ActiBioPure™ / EnzymoPure™: Internal quality designations indicating elevated purity and minimized contaminants (e.g., host-cell proteins, nucleic acids, endotoxin) to support sensitive applications; exact acceptance criteria not specified for this item; refer to CoA/Spec Sheet.
  • High Performance: Indicates performance-screened lots for robust activity under standard assay conditions (details not specified; refer to CoA/Spec Sheet).
  • Sterile: Manufactured or filtered to be free of viable microorganisms; handle aseptically to maintain sterility.

• What this means in practice:

  • Low background in kinetic assays and compatibility with sterile cell culture workflows where transient β-lactam degradation is required.
  • Consistent specific activity and minimal lot-to-lot variability (general expectation for recombinant enzymes).
  • Documentation: Certificate of Analysis (CoA) should provide protein concentration, activity units, formulation buffer, and sterility/endotoxin status; not specified here.

• Compliance: Research use only (RUO). Not intended for diagnostic or therapeutic use.

Reaction and Applications

Penicillinase (β-lactamase) catalyzes hydrolytic opening of the β-lactam ring of penicillin-class antibiotics, producing the corresponding penicilloic acids.

• Reaction type (literature):
  • Serine hydrolase mechanism: rapid acylation of the catalytic Ser-OH by the β-lactam carbonyl, followed by water-mediated deacylation to regenerate the free enzyme.
  • Substrates: benzylpenicillin (penicillin G), ampicillin, amoxicillin, and other penicillins; cephalosporin turnover depends on enzyme class/variant (literature; substrate scope is sequence-dependent).
• Research applications (non-clinical):
  • Analytical/assay reagent: remove or quantify penicillin in samples (e.g., coupled spectrophotometric assays monitoring ΔA240 nm for penicillin G).
  • Cell culture/process development: degrade β-lactam carryover in media or protein preparations when antibiotic-free conditions are required (e.g., prior to sensitive enzymology or transport studies).
  • Enzyme kinetics/structure–function: benchmark for β-lactamase inhibitor screening and mechanistic studies.
  • Sample preparation: pre-treatment step to eliminate penicillin interference before chromatographic or bioassay analysis.
• Practical tips:
  • Pre-equilibrate enzyme in assay buffer; avoid chelators/metals that may perturb stability.
  • Determine minimal effective units per mL to reach complete hydrolysis within the desired timeframe; verify by HPLC/UV.
  • For sterile workflows, add via sterile filtration or aseptic technique; remove or inactivate enzyme post-reaction as needed (e.g., heat denaturation if compatible with sample).

Note: Exact substrate specificity, turnover numbers, and activity units are enzyme-variant and formulation dependent; consult the CoA/Spec Sheet for this item.

Reaction Conditions

General literature-guided conditions for penicillin hydrolysis; actual conditions should be optimized empirically for your system.

• Substrates: Penicillin G (benzylpenicillin), ampicillin, amoxicillin; verify variant-dependent scope.
• Buffer: 25–50 mM phosphate, HEPES, or Tris, pH 7.0–8.0.
• Temperature: 25–37°C typically balances rate and stability (literature).
• Ionic strength: 50–150 mM NaCl can stabilize many enzymes.
• Enzyme loading: Adjust to achieve complete hydrolysis within 5–60 minutes depending on substrate concentration; report and track in activity units per mL (item-specific units not specified).
• Monitoring: UV at 240 nm (penicillin G shows a decrease on ring opening), HPLC (C18, aqueous-organic gradient), or LC–MS for product confirmation.
• Quench/removal: Heat denaturation (e.g., 65–80°C if sample allows), filtration, or affinity removal; verify no impact on analytes.
• Compatibility: Avoid strong oxidants, SDS, high % organic, or extreme pH which may inactivate the enzyme. Glycerol (≤10%) often tolerated; confirm for your assay.

Expected outcomes (literature): Near-quantitative conversion of penicillin to penicilloic acid under neutral pH at 25–37°C given sufficient enzyme units and mixing. Kinetic parameters (kcat, KM) are variant- and substrate-specific and are not provided for this item.

Safety and Handling

• GHS classification, signal word, pictograms, and H-statements: Not specified for this item; refer to SDS for authoritative safety information.
• General laboratory precautions (proteins/enzymes):
  • Use appropriate PPE: lab coat, nitrile gloves, safety glasses.
  • Handle in a clean area to maintain sterility and avoid enzyme aerosolization.
  • Avoid inhalation of powders/aerosols; enzymes can be respiratory/skin sensitizers in some individuals (general consideration for proteins).
• Storage incompatibilities:
  • Avoid strong oxidizers, extreme pH (<5 or >9) unless required for controlled experiments.
  • Avoid repeated freeze–thaw cycles; they can denature proteins and reduce activity.
  • Do not mix with proteases without appropriate inhibitors.
• First-aid overview (general):
  • Skin/eye contact: Rinse with water for at least 15 minutes; remove contaminated clothing; seek medical advice if irritation persists.
  • Inhalation: Move to fresh air; seek medical attention if symptoms occur.
  • Ingestion: Rinse mouth; seek medical advice.
• Spill/accidental release: Absorb aqueous solutions with inert material, avoid aerosol generation, clean with water and suitable disinfectant; dispose according to institutional and local regulations.
• Fire safety: Protein solutions are aqueous and not flammable; for packaging materials use standard extinguishing media. Thermal decomposition may produce nitrogen oxides.
• Reference: Always consult the SDS for this specific catalog item for definitive hazard classification and response measures.

Solvent Selection

As a recombinant protein enzyme, Penicillinase is used in aqueous buffers rather than organic solvents.

• Polarity and miscibility:
  • Highly water-soluble in neutral, buffered saline solutions. Avoid high concentrations of denaturing organics (e.g., >10–20% v/v acetonitrile, ethanol) unless validated for the specific application.
• Suitable buffer systems (general, literature-guided):
  • Phosphate, HEPES, MOPS, or Tris buffers, pH ~6.5–8.0.
  • Ionic strength: 50–200 mM salt (e.g., NaCl) often stabilizes proteins; optimize empirically.
  • Stabilizers: 0.1–1% carrier protein (e.g., BSA) or 5–10% glycerol can reduce adsorption and improve stability; confirm compatibility with downstream assays.
• Avoid:
  • Strong acids/bases, chaotropes (e.g., >2 M urea/guanidine), anionic detergents (SDS), heavy metal ions, and oxidants which can inactivate serine hydrolases.
  • Repeated passage through hydrophobic plastics without surfactant, which may cause adsorption losses.
• When to choose alternative media:
  • For organic-rich analytical systems, consider immobilized enzyme reactors upstream of organic eluent, or perform the hydrolysis step in aqueous buffer prior to extraction/LC.

Small comparison (general):

• Phosphate buffer (pH 7.2): robust pH control, compatible with UV assays at 240 nm.
• HEPES (pH 7.4): good biological compatibility; minimal metal chelation.
• Tris (pH 7.5–8.0): effective near alkaline pH but temperature-dependent pKa; verify assay temperature.

Storage and Reconstitution

• Storage conditions (item-specific): Store at 2–8°C.
• Shipping: Shipped on wet ice.
• Formulation and concentration: Not specified for this item; refer to CoA/Spec Sheet.
• Reconstitution: If supplied lyophilized or concentrated, reconstitute/dilute with sterile, chilled buffer (e.g., phosphate or HEPES, pH ~7.2). Gently invert to mix; avoid vigorous vortexing/foaming. Final buffer composition should match your assay and maintain sterility. Exact instructions for this item are not specified; refer to CoA/Spec Sheet.
• Handling:
  • Keep on ice during use; return promptly to 2–8°C.
  • Avoid repeated freeze–thaw; prepare single-use aliquots if freezing is necessary for your workflow (verify stability).
  • Use low-binding tubes/tips to minimize adsorption losses.
  • Maintain aseptic technique to preserve the Sterile designation.
• Stability: Short-term stability at 2–8°C is typical for proteins, but item-specific shelf life and activity retention are not specified; refer to CoA/Spec Sheet.

Research Use Only: Not for human or veterinary use. Consult the SDS and CoA for definitive guidance on formulation, stability, and activity.

Structure and Identity

Penicillinase is a β-lactamase enzyme (EC 3.5.2.6) that catalyzes hydrolysis of the β-lactam ring of penicillins to yield penicilloic acids.

• SKU: P1508160
• Product name: Penicillinase
• CAS: 9001-74-5
• Category: Protein (enzyme), recombinant; sterile preparation
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Typical literature range: ~28–35 kDa for common class A β-lactamases (literature)
• SMILES: Not applicable for proteins; not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not applicable for proteins; not specified for this item; refer to CoA/Spec Sheet.
• Structural features (general, literature):
  • Single-polypeptide enzyme typically adopting the class A β-lactamase fold (α/β domain with active-site Ser-Lys-Glu/Tyr constellation).
  • Active-site nucleophile: catalytic serine; conserved motifs e.g., SXXK, SDN, KTG (literature).
  • No defined 2D small-molecule structure; tertiary protein fold enables acylation/deacylation of β-lactam substrates.

2D description in words (general): Compact globular protein with a catalytic cleft that accommodates the β-lactam warhead of penicillins; the serine hydroxyl executes nucleophilic attack on the amide carbonyl of the strained four-membered ring (literature).

Synthetic Utility

While not a building block in the traditional organic sense, Penicillinase serves as a biocatalyst for selective bond cleavage in β-lactam antibiotics.

• Functional transformation (literature):
  • Hydrolytic opening of the strained β-lactam ring to generate penicilloic acids, preserving other functional groups under mild, aqueous conditions.
  • Selectivity allows deprotection-like removal of the β-lactam “warhead” without affecting side chains that might be labile under acid/base hydrolysis.
• Uses in synthesis/analytical chemistry:
  • Preparation of reference penicilloic acid standards for method development.
  • Cleanup step in multicomponent mixtures to remove penicillin prior to downstream reactions or analyses.
  • Coupled-enzyme or chemical assays exploiting the UV spectral change upon β-lactam hydrolysis (ΔA240 nm) for kinetic calibration.
• Retrosynthetic value:
  • Enables orthogonal disconnection of the β-lactam amide under biocompatible conditions, complementing chemical hydrolysis routes.
• Practical guidance:
  • Optimize enzyme units relative to substrate load and matrix effects (proteins, salts).
  • Consider immobilized preparations for flow or reusability.
  • Validate completeness of hydrolysis by LC/UV/MS; quench or remove enzyme before subsequent steps to prevent unintended reactions.

Item-specific activity, turnover rates, and immobilization compatibility are not provided; consult CoA/Spec Sheet or perform pilot screening.

Target Specificity

Not applicable. This product is an enzyme, not an antibody or affinity reagent. No target/epitope specificity, clone, isotype, or species reactivity information is provided for this item.