Zymolyase, CAS No.37340-57-1

CAS: 37340-57-1 Cat. No.: Z1523272 EG-Nummer: 609-409-4

GRADE & PURITY Bioactive, ActiBioPure™, Native, High Performance, EnzymoPure™, from Arthrobacter luteus; ≥100,000 U/g enzyme powder

Übersicht

Zymolyase, produced by a submerged culture of Arthrobacter luteus, has strong lytic activity against living yeast cell walls, to produce protoplast or spheroplast of various strains of yeast cells. An essential enzyme for the lytic activity of Zymolyase is β-1,3-glucan laminaripentaohydrolase. It hydrolyzes linear glucose polymers with β-1,3-linkages and releases specifically laminaripentaose as the main and minimum product unit. Essential enzyme: β-1,3-glucan laminaripentaohydrolase ptimum pH and Temp.: pH7.5, 35°C (for lysis of viable yeast cells) pH6.5, 45°C (for hydrolysis of yeast glucan) Stability (2°C): No loss of activity was found after storage for 1 year Specificity (Lytic Spectrum) : Ashbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloeckera, kluyveromyces, Lipomyces, Metschnikowia, Pichia, Pullularia, Torulopsis, Saccharomyces, Saccharomycopsis, Saccharomycodes, Schwanniomyces, etc. Unit Definition One unit of lytic activity is defined as that amount which indicates 30% of decrease in absorbance at 800 nm(A800) of the reaction mixture under the following condition. Reaction Mixture Enzyme solution: 1 ml (0.012-0.024 mg/ml for Zymolyase) Brewer's yeast cell suspension: 3 ml (2 mg/ml) 1/15M Phosphate buffer: 5 ml (pH7.5) Distilled water: 1 ml After incubation for 2 hours at 25°C with gentle shaking, A800 of the mixture is determined. When 60% of A800 decrease, equivalent to 2 units, is observed in the reaction system, the brewer’s yeast cells are completely lysed, namely 1 unit of Zymolyase lyses 3 mg dry weight of brewer’s yeast. Preparation of 1/15 M Phosphate Buffer (1L of pH 7.5 buffer) Dissolve 19.1 g of di-sodium hydrogenphosphate 12-hydrate in distilled water to make an 800 mL solution. Dissolve 1.814 g of potassium dihydrogenphosphate in distilled water to make a 200 mL solution. Mix the two solutions.

Specifications

Product Name

Zymolyase, CAS No.37340-57-1

Note

ActiBioPure™, Bioactive, High Performance, Native, EnzymoPure™

Spezifikationen & Reinheit

ActiBioPure™,Bioactive,High Performance,EnzymoPure™,Native,from Arthrobacter luteus; ≥100,000 U/g enzyme powder

CAS

37340-57-1

Molekültyp

Enzyme

Product Line

Biologische Produkte

Lagerung und Versand

Konzentration

from Arthrobacter luteus; ≥100,000 U/g enzyme powder

Storage

Store at 2-8°C

Verschickt in

Wet ice

Application Protocols

Representative literature protocols are provided for guidance only. Optimize for your strain and consult the CoA/Spec Sheet for activity definitions.

A. Yeast spheroplasting (literature example)

• Grow cells to mid‑log phase; harvest 1–5 × 10^8 cells; wash once in 1.0 M sorbitol.
• Resuspend in 1.0 M sorbitol, 50 mM potassium phosphate, pH 7.4 (optional 10 mM EDTA for nucleic‑acid workflows).
• Add Zymolyase to 0.2–2 U/mL starting dose; incubate at 30°C with gentle rocking for 15–45 min.
• Monitor by OD800 decrease or hypotonic lysis test. When endpoint is reached, place on ice and wash twice with cold 1.0 M sorbitol buffer.
• Proceed to transformation, microscopy, or lysis as required.

B. DNA extraction from yeast (literature example)

• Pre‑digest cell pellets in 1.0 M sorbitol, 50 mM Tris‑HCl pH 7.5, 10 mM EDTA with Zymolyase (0.5–2 U/mL) at 30°C for 15–30 min.
• Pellet spheroplasts gently; resuspend in lysis buffer (e.g., 1% SDS, 100 mM NaCl, 50 mM Tris‑HCl pH 8.0, 10 mM EDTA). Proceed with proteinase K, RNase, and phenol‑free or phenol‑chloroform cleanup per institutional SOP.

C. Permeabilization for immunostaining (literature example)

• Fix cells if required; wash; treat briefly with dilute Zymolyase in isotonic buffer (e.g., 0.1–0.5 U/mL, 10–15 min) to increase antibody access, avoiding over‑digestion.

These are general literature procedures and not product‑specific recommendations.

Biological Roles

Zymolyase is a preparation of microbial enzymes that hydrolyze β‑1,3‑glucans—key structural polysaccharides of many yeast cell walls (literature). While not a single defined biomolecule, its constituents have the following biological context:

• Source and physiology (literature, general): Often derived from Arthrobacter species, where glucanases participate in environmental carbohydrate turnover and nutrient acquisition.
• Substrate specificity: Targets β‑1,3‑linked D‑glucose polymers (glucans) with occasional action on branching points depending on accessory activities; catalysis yields oligosaccharides and glucose.
• Role in experimental systems: Enables controlled perturbation of the yeast cell envelope, facilitating studies of membrane physiology, morphogenesis, secretion, and cell‑wall assembly.
• Impact on cells (literature, general): At appropriate doses in osmoticum, removes the wall without rupturing the plasma membrane, yielding viable spheroplasts that can regenerate walls under recovery conditions.
• Interactions with biomolecules: By degrading glucan, it increases accessibility of intracellular proteins, nucleic acids, and organelles to probes and extraction reagents while minimizing shear damage.

Note: The above describes general biological roles of zymolyase‑type enzymes; item‑specific composition and activity spectra vary. For exact characteristics of this product, consult the CoA/Spec Sheet.

Buffer Applications

Zymolyase is typically used in buffered osmotic stabilization media to prevent hypotonic lysis during cell‑wall digestion (literature, general). Below are commonly reported systems; adapt empirically for your strain and application.

• Spheroplasting buffer (literature example):
  • 1.0 M sorbitol, 10–50 mM potassium phosphate, pH 7.2–7.5; optional 10 mM EDTA to inhibit nucleases when subsequent nucleic‑acid extraction is planned.
  • Alternative buffers: 1.0 M sorbitol + 50 mM Tris‑HCl (pH 7.5) or 1.0 M sorbitol + 50 mM HEPES (pH 7.2).
• Wash/holding buffer: Same osmotic strength (0.8–1.2 M sorbitol) with 10–50 mM buffer; keep on ice or at room temperature according to downstream needs.
• Additives and compatibility (literature, general):
  • Mg2+/Ca2+: Not required for catalytic activity in many preparations; may be present for downstream steps (e.g., transformation). If chelators are used, verify compatibility with your workflow.
  • Detergents: Avoid during digestion; introduce mild nonionic detergents only after spheroplasting if needed for lysis.
  • Protease/nuclease inhibitors: Optional; ensure they do not inhibit the enzyme or downstream enzymes.
• Sterility: Filter‑sterilize buffers (0.22 µm) and maintain aseptic technique for viability‑critical workflows (e.g., transformation or regeneration assays).

These are literature‑style recipes for guidance and are not product specifications. Optimize concentrations and pH for your yeast species and growth conditions.

Green Alternatives

Enzymatic cell‑wall digestion with Zymolyase is itself a greener alternative to harsh chemical or energy‑intensive lysis methods (literature, general).

Comparison (literature/general values):

• Approach: Zymolyase (enzymatic)

  • Pros: Aqueous buffers; low energy input; selective for β‑1,3‑glucan; preserves macromolecular complexes; reduced aerosol and microplastic generation vs bead‑beating.
  • Cons: Biologic sourcing; cold‑chain required; activity can vary with matrix and lot—requires titration.

• Approach: Alkaline/chemical lysis (e.g., NaOH/SDS preps)

  • Pros: Inexpensive, rapid for bulk DNA minipreps.
  • Cons: High pH/denaturants create hazardous waste; damages proteins/organelles; can shear nucleic acids; less selective.

• Approach: Mechanical disruption (bead‑beating, French press)

  • Pros: Reagent‑minimal; broadly applicable.
  • Cons: High energy consumption; heat generation; equipment wear; aerosolization; less gentle on targets.

Greener practice tips with Zymolyase (literature, general):

• Use minimal effective enzyme dose and recycle osmotic buffers when compatible with your workflow (filter/sterilize for reuse).
• Replace NaN3 with sterile technique or mild biocides in short‑term stocks where permissible.
• Consolidate incubations at 30°C to reduce energy relative to 37°C when performance allows.
• Dispose of proteinaceous and microbial waste via institutional biological routes; avoid unnecessary solvent use.

Pharmaceutical Uses

This product is provided for research use only. It is not intended for use as an active pharmaceutical ingredient or in human/animal therapies.

• Possible roles in pharmaceutical R&D workflows (literature, general):
  • Sample preparation enzyme for analytical characterization of yeast or fungal bioprocess organisms.
  • Tool enzyme to generate spheroplasts for cell‑based assays and screening.
  • Pre‑treatment step to facilitate extraction of nucleic acids or intracellular targets from yeast used in process development.
• Regulatory status: No pharmacopeial monograph is implied by this listing. Any GMP use would require independent qualification, validation, and regulatory review.

No clinical or therapeutic claims are made or should be inferred.

Physical Properties

Item‑specific physical constants are not applicable to enzyme mixtures. Where relevant, consult the CoA.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Protein nature: Heterogeneous enzyme preparation (literature, general).
• Solubility (literature, general):
  • Readily soluble in common biological buffers (e.g., phosphate, Tris, HEPES) and isotonic osmotic stabilizers (e.g., 0.8–1.2 M sorbitol) used for yeast spheroplasting.
  • Insoluble/denatured by most organic solvents (e.g., high % ethanol, acetone) and by anionic detergents (e.g., SDS) at denaturing concentrations.
• pH activity profile (literature, general): Typically active near neutral to slightly alkaline pH (~6.5–8). Optimal pH depends on exact composition and should be empirically determined for the lot.
• Temperature profile (literature, general): Functional between ~20–37°C in cell‑wall digestion workflows; excessive heat (>45–50°C) diminishes activity.
• Stabilizers (general guidance): Activity often preserved by inclusion of osmolytes (sorbitol), glycerol (≤50% v/v), BSA (≤0.1%), and avoidance of repeated freeze–thaw.

Important: Do not treat the above as specifications; they are general literature expectations for zymolyase‑type enzyme preparations. For lot‑specific values and formulation, refer to the product CoA/Spec Sheet.

Quality and Grades

This item is offered under multiple performance‑oriented research grades. No additional item‑specific specifications (e.g., activity units, contaminants) are provided here; refer to CoA/Spec Sheet for lot data.

• Grades listed for this item (as provided):
  • Bioactive
  • ActiBioPure™
  • Native
  • High Performance
  • EnzymoPure™

What these grades generally imply (literature/general definitions; not item‑specific specs):

• Bioactive: Supplied to retain catalytic activity under recommended storage/handling; suitable for functional workflows (e.g., spheroplasting, yeast lysis).
• ActiBioPure™: Emphasis on high activity recovery and minimized non‑enzymatic components to reduce background in downstream assays.
• Native: Sourced/maintained in a form closely reflecting the natural enzyme mixture (non‑recombinant), preserving native post‑translational features.
• High Performance: Tight process controls for consistent activity and low lot‑to‑lot variability aimed at demanding workflows.
• EnzymoPure™: Additional polishing or impurity reduction to lower protease/nuclease backgrounds and host‑derived contaminants, improving compatibility with nucleic‑acid workflows and proteomics.

Practical considerations:

• Lower background protease/nuclease burden is advantageous for intact RNA/DNA isolation and for spheroplasting prior to transformation or microscopy.
• Verify activity on your strain and matrix; yeast cell wall composition varies with species, growth phase, and conditions. For quantitative needs, consult activity units and assay method on the CoA.

Reaction and Applications

Zymolyase is employed for selective hydrolysis of β‑1,3‑glucan in yeast and fungal cell walls (literature). Typical application families include:

• Yeast spheroplasting: Controlled removal of the cell wall while maintaining plasma membrane integrity for downstream transformation, fusion, patch‑clamp, or microscopy.
• Nucleic acid extraction: Pre‑digestion of the cell wall to facilitate DNA/RNA isolation under mild conditions, often combined with EDTA and detergents post‑digestion.
• Organelle isolation: Gentle lysis improves recovery of nuclei, mitochondria, ER, and complexes compared to mechanical shear.
• Cell biology assays: Permeabilization for immunostaining, in situ hybridization, and reporter access in intact cells.
• Polysaccharide analysis: Generation of defined β‑1,3‑glucan oligosaccharides for analytical workflows.

Practical tips (literature, general):

• Start with 0.1–5 U/mL equivalent activity at 30–37°C for 10–60 min in 1 M sorbitol buffer, pH 7.2; optimize for strain, growth phase, and temperature.
• Monitor spheroplasting by OD800 decrease in osmoticum or by staining (e.g., Calcofluor White resistance loss); stop the reaction by chilling and washing in sorbitol buffer.
• Maintain osmotic stabilization (≥0.8 M sorbitol) at all times until cell wall regeneration or downstream lysis is complete.
• For nucleic acid prep, follow digestion with gentle detergent lysis (e.g., SDS or Triton X‑100 at appropriate steps) only after cell wall removal.

Note: Activity units, specific activity, and exact buffer formulation are preparation‑dependent; refer to CoA/Spec Sheet for item‑specific details.

Reaction Conditions

Typical working conditions for Zymolyase‑type preparations (literature, general guidance; optimize empirically and consult the CoA for your lot):

• Substrate: Intact yeast/fungal cells with β‑1,3‑glucan walls (log‑phase cells are generally more susceptible than stationary phase).
• Buffer: 10–50 mM phosphate/Tris/HEPES at pH 7.0–7.5 with 0.8–1.2 M sorbitol for osmotic stabilization.
• Temperature: 30–37°C for 10–60 min; cooler temperatures slow reactions but can improve viability.
• Enzyme dosing: Common starting range 0.1–5 U/mL; adjust based on strain, cell density, and desired extent of wall removal. Units and assay definitions vary by supplier—align to CoA.
• Agitation: Gentle inversion/rocking to keep cells suspended; avoid shear that ruptures spheroplasts.
• Quench/stop: Chill on ice and wash with ice‑cold osmotic buffer to halt digestion.
• Monitoring endpoints:
  • Turbidity drop at 800 nm in presence of osmoticum (literature assay).
  • Sensitivity to hypotonic shock: spheroplasts lyse in water, indicating successful wall removal.
  • Microscopy with wall stains (e.g., Calcofluor White) to confirm loss of glucan signal.
• Expected outcomes: Partial to complete spheroplasting with preserved membrane integrity; yield depends on strain and growth conditions.

Caveats: High ethanol, detergents (SDS), or extremes of pH/temperature reduce activity. Presence of endogenous proteases can diminish enzyme performance—consider cold handling and additives compatible with your workflow.

Safety and Handling

• GHS classification, signal word, pictograms, H‑statements: Not specified for this item; refer to SDS.
• General hazards (literature, general): Proteinaceous powders/aerosols may cause allergic sensitization in susceptible individuals. Avoid inhalation and skin/eye contact. Not intended for human or animal use.
• Recommended PPE: Laboratory coat, nitrile gloves, safety glasses or face shield when splashes/aerosols are possible. Use a biological safety cabinet when handling dry powders or preparing concentrated stocks to minimize aerosols.
• Handling guidance:
  • Prepare solutions gently; avoid vigorous vortexing that generates foam/aerosols.
  • Maintain cold chain as practical to preserve catalytic activity.
  • Avoid contamination with proteases or harsh detergents (SDS) that denature enzymes.
  • Segregate from strong oxidizers and acids/bases that can denature proteins.
• First‑aid overview (literature, general):
  • Inhalation: Move to fresh air; seek medical advice if symptoms persist.
  • Skin: Wash with soap and water; remove contaminated clothing.
  • Eyes: Rinse cautiously with water for several minutes; remove contact lenses if present and easy to do.
  • Ingestion: Rinse mouth; seek medical advice.
• Spills and disposal: Absorb aqueous spills with inert material; avoid dust formation. Dispose according to institutional and local regulations for proteinaceous laboratory waste.

Always consult the product SDS for authoritative safety information.

Solvent Selection

As a protein enzyme preparation, Zymolyase is not used with organic solvent systems; performance depends on aqueous buffer composition.

• Preferred media (literature, general):
  • Isotonic buffers with osmotic stabilizers for yeast spheroplasting (e.g., 0.8–1.2 M sorbitol with phosphate, Tris, or HEPES at pH ~7.0–7.5) to prevent osmotic lysis during cell‑wall removal.
  • Salt conditions: 0–200 mM monovalent salts (KCl/NaCl) are typically tolerated; high ionic strength may reduce activity and cell integrity.
  • Avoid strong denaturants (SDS, urea >2–3 M), high % organic solvents, and oxidizing conditions.
• Additives (literature, general):
  • EDTA (1–10 mM) is often included to chelate divalent metals and inhibit endogenous nucleases when extracting nucleic acids from yeasts; compatibility should be verified for the specific preparation.
  • Reducing agents (e.g., 1–5 mM DTT or β‑ME) are generally not required and may affect protein stability; use only if justified by the workflow.

Comparison to alternatives (literature, general):

• Zymolyase vs lyticase: Both target β‑1,3‑glucan; activity profiles and side activities differ by supplier/preparation—empirical titration is recommended.
• Zymolyase vs mechanical lysis (bead‑beating): Enzymatic approach is gentler, preserves organelles and macromolecular complexes, and enables spheroplasting for transformation/microscopy; mechanical lysis is faster but harsher and can shear nucleic acids.

Storage and Reconstitution

• Storage conditions (as provided): Store at 2–8°C. Shipped on wet ice. Maintain cold chain to preserve activity.
• Form and appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Reconstitution (literature, general):
  • Dissolve at the desired activity/concentration in a compatible buffer (e.g., 10–50 mM potassium phosphate, pH ~7.0–7.5) with osmotic stabilizer as needed for immediate use.
  • For working stocks, 10–50% glycerol can improve stability; filter‑sterilize (0.22 µm) if sterility is required.
  • Avoid vigorous mixing; gently swirl to minimize foaming.
• Aliquoting and freeze–thaw (literature, general):
  • Minimize freeze–thaw cycles; prepare single‑use aliquots. Many users store aliquots at −20°C or −80°C for extended periods; validate stability for your application and follow your institution’s best practices.
• In‑use stability (literature, general): Keep on ice during setup; warm only for the incubation step. Discard solutions showing microbial growth or activity loss.
• Compatibility: Avoid strong oxidants, extremes of pH, high organic solvent content, and denaturing detergents during storage.

For lot‑specific formulation, concentration, and activity units, consult the CoA/Spec Sheet. This product is for research use only.

Structure and Identity

Zymolyase is a protein enzyme preparation widely used to hydrolyze β-1,3‑glucan in yeast cell walls, producing spheroplasts and facilitating gentle lysis (literature). As a biologic mixture, it does not have a single small‑molecule structure.

• SKU: Z1523272
• Product name: Zymolyase
• CAS: 37340-57-1
• Category: Protein (enzyme 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.
• SMILES: Not applicable to enzyme mixtures; not specified for this item.
• InChIKey: Not applicable to enzyme mixtures; not specified for this item.
• Structural features (literature, general):
  • Mixture of cell‑wall–degrading enzymes; principal activities commonly include β‑1,3‑glucan laminaripentaohydrolase and β‑1,3‑glucanase that cleave β‑1,3‑glucan backbones, often with minor mannanase/chitinase activities depending on preparation.
  • Proteinaceous, typically glycosylated bacterial enzymes sourced from Arthrobacter spp. (literature).
  • No defined 2D small‑molecule depiction; functionally characterized by substrate specificity for β‑1,3‑glucans.

2D description in words (literature, general): Enzyme polypeptides with catalytic domains that recognize β‑1,3‑linked D‑glucose polymers; active sites accommodate glucan chains and perform endo‑ or exo‑type hydrolysis, yielding oligosaccharides and glucose.

Synthetic Utility

Although not a chemical building block, Zymolyase functions as a biocatalyst enabling specific transformations and preparative steps (literature, general):

• Biocatalysis: Selective hydrolysis of β‑1,3‑glucan linkages in polysaccharides, generating soluble oligosaccharides for analysis or feedstocks for further enzymatic processing.
• Enabling chemistry: Facilitates access to intracellular enzymes and metabolites in yeast for biotransformations by removing the mass‑transfer barrier of the cell wall while preserving metabolic function in spheroplasts.
• Workflow integration:
  • Prior to chemical derivatization of intracellular metabolites or proteins, enzymatic wall removal yields cleaner extracts with reduced mechanical debris.
  • In synthetic biology, spheroplasts can accept DNA via PEG‑mediated fusion, supporting genome engineering pipelines (literature).
• Compatibility considerations:
  • Maintain osmoticum during and after digestion to keep spheroplasts intact for downstream biocatalytic assays.
  • Avoid oxidants and denaturants that can inactivate both Zymolyase and target intracellular catalysts.

This section summarizes typical uses of glucanase enzyme preparations and does not constitute item‑specific performance specifications.

Target Specificity

This section is primarily relevant to antibodies or affinity reagents; Zymolyase is an enzyme preparation and does not have an antigen/epitope specificity in that sense.

• Functional specificity (literature, general): Preferential hydrolysis of β‑1,3‑glucan linkages in yeast cell walls, with activity profile dependent on preparation.

No clone, isotype, epitope, or species‑reactivity data apply to this product.