β-Fructosidase , CAS No.37288-56-5

CAS: 37288-56-5 Cat. No.: F1444988

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Descripción general

β-Fructosidase is an enzyme that hydrolyzes sucrose to release fructose and glucose.

Specifications

Condiciones de almacenamiento de almacenamiento

Store at -20°C

Enviado en

Ice chest + Ice pads

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Application Protocols

Only application protocols provided in the Product Data may be listed here.

Item-specific protocols: Not specified for this item; refer to CoA/Spec Sheet and SDS for any recommended assay conditions, unit definitions, or handling instructions.

General protocols are available in the Reaction Conditions tab (literature guidance) but are not item-specific directions.

Biological Roles

Literature overview:

• Physiological function: β-Fructosidases are widespread in plants, fungi, and bacteria, where they mobilize carbon by hydrolyzing sucrose and fructans. In plants, cell wall and vacuolar invertases regulate phloem unloading, sink strength, and developmental processes by controlling sucrose partitioning. In microbes (e.g., Saccharomyces, Aspergillus), secreted invertases enable assimilation of sucrose and raffinose-family oligosaccharides.
• Mechanism: Retaining glycoside hydrolases (commonly GH32/GH68) utilize a double-displacement mechanism via a covalent fructofuranosyl-enzyme intermediate, yielding net retention at the anomeric center of fructose. High substrate concentrations favor transfructosylation, producing short-chain fructooligosaccharides.
• Regulation and localization: Many eukaryotic β-fructosidases are glycoproteins targeted to the cell wall or secretory pathway; expression is induced by sucrose and repressed by glucose (catabolite repression). Bacterial enzymes may be periplasmic or extracellular, sometimes associated with PTS systems.
• Substrate specificity: While sucrose is the prototypical substrate, many enzymes also act on raffinose, stachyose, and inulin-type fructans (exo-activity), with specificity modulated by subsite architecture.
• Inhibition/activation: Heavy metals and strong oxidants typically inhibit activity; some enzymes are stabilized by calcium or neutral salts, though they are not metal-dependent. Glucose/fructose product inhibition is common and relevant for process design.

Note: This section summarizes general biological roles; the organismal source and specific biochemical properties of this catalog item are Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

β-Fructosidase is not a buffering reagent; however, assay performance depends critically on buffer selection.

General/literature guidance for enzyme assays:

• pH range: Many preparations show maximal activity in acidic buffers (pH 4.5–5.5 for yeast/fungal enzymes). Select buffer systems with minimal temperature dependence and ionic interactions.
• Common buffers: 50 mM sodium acetate (pH 4.8–5.2) or 50 mM citrate–phosphate (pH 4.5–6.0) are widely used. MES (pKa 6.1) can support slightly higher pH assays.
• Ionic strength: 10–100 mM total buffer concentration is typical; higher salt may reduce activity.
• Additives: 0–10% glycerol can stabilize in kinetic assays; EDTA (0.1–1 mM) may mitigate trace-metal inhibition. Avoid strong detergents and oxidants.
• Example recipe (literature, illustrative): 50 mM sodium acetate, pH 5.0; 0.1 mM EDTA; 0.02% sodium azide (if long assay times and compatible with downstream analytics). Always verify compatibility with detection method (e.g., DNSA reducing sugar assay, GOD–POD enzymatic readout, or HPAEC-PAD).

Not applicable use cases: β-Fructosidase is not used to prepare biological buffers per se; instead, choose buffers to support its catalytic function. Item-specific buffer compatibility or formulation details are Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Enzymatic β-fructofuranoside hydrolysis/synthesis offers a greener alternative to traditional acid-catalyzed carbohydrate transformations.

Environmental and process comparison (literature-general):

• Catalyst nature: • β-Fructosidase: Biodegradable, nontoxic protein catalyst; operates in water at mild temperatures. • Mineral acids (e.g., HCl, H2SO4): Corrosive, hazardous; require neutralization and generate salt waste.

• Conditions: • Enzyme: pH typically 4.5–6.0, 25–60°C; aqueous media with low energy input. • Acid: pH <1, elevated temperatures for rapid rates; potential for sugar degradation and coloring (caramelization, HMF formation).

• Selectivity: • Enzyme: High regio- and stereoselectivity; tunable transfructosylation to form specific FOS structures. • Acid: Poor selectivity; side reactions yield humins and byproducts.

• Workup: • Enzyme: Simple filtration (especially when immobilized), minimal neutralization; reusable when immobilized. • Acid: Neutralization, corrosion control, and additional purification steps.

Tradeoffs and considerations:

• Enzymes can be costlier on a per-weight basis and may exhibit limited solvent tolerance and thermal stability; immobilization and process engineering mitigate these constraints.
• For harsh feedstocks or high-solids slurries, pretreatment and enzyme-compatible pH adjustment are required; acid catalysis may remain advantageous where extreme conditions are unavoidable.

Conclusion: For lab- and pilot-scale carbohydrate transformations, β-fructosidase is typically the greener, safer choice with superior selectivity and reduced waste.

Pharmaceutical Uses

No therapeutic claims are made; this product is for research use only.

General roles in pharmaceutical and bioprocess R&D (literature):

• Biocatalysis in process development: β-Fructosidase can serve as a selective catalyst for hydrolysis of β-D-fructofuranosidic protecting groups in carbohydrate intermediates under mild, aqueous conditions.
• Analytical support: Enzymatic conversion of sucrose-containing excipients to monosaccharides for quantitative assays (e.g., LC, IC), facilitating impurity profiling or stability studies.
• Prebiotic carbohydrate synthesis: Transfructosylation to generate defined fructooligosaccharides (FOS) for research on excipient interactions, microbiome-model media, or formulation screening.
• Immobilized enzyme reactors: Continuous-flow generation of invert sugar or FOS for formulation development studies; immobilization enhances operational stability and simplifies separation from product streams.
• Compatibility considerations: Stabilizers, preservatives, or carrier proteins in the enzyme preparation (if present) may interfere with downstream analytics (e.g., MS ion suppression). Verify on the Spec Sheet before use in regulated workflows.

Item-specific regulatory or compendial status (e.g., USP/NF, Ph. Eur.), residual solvent limits, bioburden/endotoxin levels, and excipient acceptability are Not specified for this item; refer to CoA/Spec Sheet. All applications should be validated within the intended quality system.

Physical Properties

As a protein enzyme preparation, conventional small-molecule physical constants (bp, mp, logP, refractive index) are not applicable.

Item-specific physical data:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight (of preparation or specific isoform): Not specified for this item; refer to CoA/Spec Sheet.

General/literature characteristics (will vary with source and formulation):

• Typical monomer size: 55–95 kDa per subunit for many GH32 β-fructofuranosidases; some enzymes are secreted glycoproteins and may be monomeric or oligomeric (literature).
• Isoelectric point (pI): Broadly variable (pI ~4–5.5 common for yeast/fungal forms), influenced by glycosylation and sequence (literature).
• Solubility: Readily soluble in aqueous buffers; activity retained in mildly ionic media. High organic cosolvent (>10–20% v/v) generally reduces activity; 10–50% glycerol often stabilizes during storage (literature).
• Optimal pH: Often acidic, pH 4.5–5.5 for yeast/fungal invertases; some bacterial forms show neutral optima (literature).
• Optimal temperature: Commonly 40–60°C depending on source; thermal stability varies widely (literature).
• Metal ion effects: Activity typically independent of metal cofactors; heavy metals (e.g., Hg2+, Ag+) can be inhibitory (literature).
• Kinetic parameters: Km for sucrose typically in the low to mid millimolar range; kcat and specificity depend strongly on enzyme source and assay (literature).

Note: Actual activity units, specific activity, formulation (buffer, salts, stabilizers), moisture, and protein content are Not specified for this item; refer to CoA/Spec Sheet and SDS for definitive values.

Quality and Grades

Item-specific quality information:

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
• Specific activity, unit definition, protein content, stabilizers/preservatives: Not specified for this item; refer to CoA/Spec Sheet.

General guidance on enzyme quality attributes (literature/industry practice):

• Activity units: β-Fructosidase is typically specified in “Units (U),” where 1 U liberates 1 µmol fructose (or reducing sugar equivalents) per minute from sucrose under defined conditions (e.g., pH ~4.5–5.5, 37–55°C). Always confirm the exact unit definition on the CoA, as assay pH/temperature strongly impact reported activity.
• Purity descriptors: Terms such as “crude,” “partially purified,” or “≥x U/mg protein” indicate enrichment level; protease-free and nucleic-acid-free grades are preferred for molecular biology contexts to minimize adventitious degradation.
• Endotoxin/bioburden: For cell-based work, low bioburden and low endotoxin are desirable; not usually specified for bulk biocatalysis.
• Stabilizers: Enzyme formulations may include salts, sugars, BSA, or polyols (e.g., glycerol) to enhance stability. Presence of any stabilizer or preservative should be verified on the Spec Sheet; this affects compatibility with downstream assays (e.g., mass spectrometry, HPLC).
• Batch-to-batch consistency: For kinetic work, match the same lot or normalize by specific activity. Always record activity units per reaction rather than mass of material.
• QC testing: Typical tests include activity assay on sucrose, SDS-PAGE profile, residual protease assay, and microbial limits, as applicable.

Reaction and Applications

Core catalytic function (literature): β-Fructosidase (invertase; EC 3.2.1.26) catalyzes hydrolysis of terminal, nonreducing β-D-fructofuranosidic linkages. Principal reaction examples:

• Sucrose → D-glucose + D-fructose (invert sugar)
• Raffinose → melibiose + D-fructose
• Fructans/inulin-type oligomers → fructose (exo-inulinase activity in some enzymes)

Hydrolysis versus synthesis:

• Retaining mechanism enables transfructosylation at high substrate concentrations, forming fructooligosaccharides (FOS; e.g., 1-kestose, nystose) valued as prebiotic carbohydrates. Reaction outcome is tunable by water activity, substrate level (0.5–2.5 M sucrose), temperature, and enzyme loading (literature).

Research and process applications (literature):

• Carbohydrate analysis: Generation of monosaccharides for HPAEC-PAD or GC-MS profiling; enzymatic confirmation of linkage types.
• Biocatalysis: Selective cleavage of β-fructofuranosides in protecting-group strategies; synthesis of defined FOS via transfructosylation.
• Food/bioprocess modeling: Bench-scale production of invert sugar for rheology and fermentation studies; saccharification of raffinose-family oligosaccharides to mitigate anti-nutritional factors in plant matrices.
• Immobilization: Enzyme immobilized on silica, polymers, or magnetic supports improves operational stability, reuse, and compatibility with continuous flow.

Practical tips (general):

• Maintain pH near the enzyme’s optimal range (often 4.5–5.5 for yeast/fungal preparations). Buffer capacity should accommodate acid/base shifts from substrate/product.
• Control water activity to bias hydrolysis (high water) vs transfructosylation (reduced water, high sucrose).
• Avoid heavy-metal contamination and strong oxidants; pre-filter crude feeds to reduce fouling and proteolysis.

Reaction Conditions

The following are general, literature-based conditions for β-fructosidase; optimize for your specific enzyme source and formulation. Item-specific activity, unit definitions, and stabilizers are Not specified for this item; refer to CoA/Spec Sheet.

Typical hydrolysis of sucrose:

• Substrate: 10–200 mM sucrose in 50 mM sodium acetate, pH 5.0.
• Enzyme loading: 0.01–1 U per mL reaction (adjust to desired rate).
• Temperature: 30–55°C (commonly 37–50°C for yeast/fungal enzymes).
• Time: 5–60 min for analytical conversions; 1–12 h for preparative scales.
• Monitoring: DNSA reducing sugar assay, GOD–POD enzymatic glucose assay, or HPAEC-PAD for sugar profiles.

Transfructosylation to FOS (favoring synthesis):

• Substrate: 1.0–2.5 M sucrose to reduce water activity.
• Buffer: 50–100 mM acetate or citrate–phosphate, pH 5.0–5.5.
• Enzyme loading: 1–20 U per mL (depends on target DP and rate).
• Temperature: 40–60°C, balancing activity vs stability.
• Time: 1–24 h; quench by heating (e.g., 80–90°C, brief) or pH shift, then desalt and purify by chromatography.

Compatibility/inhibitors:

• Heavy metals (Hg2+, Ag+) and strong oxidants inhibit activity; EDTA (0.1–1 mM) often protective.
• Organic cosolvent tolerance is limited; keep ≤10–20% v/v unless using specially engineered/immobilized forms.

Immobilized enzyme operations:

• Packed-bed at 30–50°C with 10–60 min residence time can provide steady conversion; monitor pressure drop and fouling.

All values above are literature guidance; verify with small-scale trials.

Safety and Handling

Authoritative safety information must be taken from the product SDS. The following are general considerations for enzyme powders/solutions and β-fructosidase preparations.

Item-specific hazard data from Product Data:

• Signal Word: Not specified for this item; refer to SDS.
• H-Statements: Not specified for this item; refer to SDS.
• GHS Classification/Pictograms: Not specified for this item; refer to SDS.

General precautions (literature/industry practice):

• Potential hazards: Enzyme proteins can be respiratory sensitizers; avoid aerosolization and inhalation. May cause skin/eye irritation in susceptible individuals.
• PPE: Lab coat, nitrile gloves, safety glasses; use a fume hood or biosafety cabinet to minimize aerosols when handling dry powders or vigorous pipetting.
• Handling: Avoid creating dust. For solutions, minimize foaming; surfactants can denature enzymes. Keep cold except during brief handling to maintain activity.
• Incompatibilities: Strong oxidizers, extremes of pH, heavy metal ions (e.g., Hg2+, Ag+), and organic solvents can denature/inhibit enzyme activity (general guidance).
• First aid (overview; defer to SDS): If inhaled, move to fresh air and seek medical attention for persistent symptoms. In case of skin contact, wash with soap and water. For eye exposure, rinse cautiously with water for several minutes; remove contact lenses if present and easy to do. If ingested, rinse mouth; seek medical advice.
• Spill response: Avoid dust generation; gently dampen and absorb with inert material. Dispose according to institutional and local regulations.
• Fire safety: Proteinaceous materials are combustible; use standard extinguishing media (CO2, foam, dry chemical). Thermal decomposition can produce nitrogen oxides and carbon oxides.

Solvent Selection

This product is an aqueous-enzyme reagent rather than a solvent; selection pertains to buffer systems and cosolvents that preserve catalytic activity.

General/literature guidance:

• Primary medium: Aqueous buffers. Acetate, citrate-phosphate, or MES buffers are commonly used near the acidic pH optimum (pH 4.5–6.0), depending on enzyme source.
• Ionic strength: 10–100 mM buffer is typical; excessive ionic strength (>0.5 M) can reduce activity or alter substrate binding.
• Cosolvents: 10–50% v/v glycerol is often used for storage stability at −20°C; low percentages of ethanol, DMSO, or PEG may be tolerated but generally lower catalytic rates. Validate tolerances empirically if substrates require limited organic content.
• Detergents: Nonionic detergents (e.g., 0.01% Tween-20) can reduce nonspecific adsorption in some assays but may destabilize certain preparations; evaluate case by case.
• Metal ions and additives: No metal cofactor requirement is typical. Chelators (EDTA 0.1–1 mM) are often compatible and can mitigate trace-metal inhibition; however, some formulations may already contain stabilizers—verify on Spec Sheet.

When to choose alternatives:

• For neutral/alkaline pH processes or substrates incompatible with acid conditions, consider fructosidases from bacterial sources with higher pH optima, or alternative hydrolases (e.g., α-glucosidase for α-linkages). For robust organic media, immobilized β-fructosidase or engineered variants may offer improved solvent tolerance (literature).

Storage and Reconstitution

Item-specific storage and shipping (from Product Data):

• Storage Conditions: Store at -20°C.
• Shipped In: Ice chest + Ice pads.

Additional general guidance (literature/practice; verify against Spec Sheet for compatibility):

• Short-term handling: Keep on ice during use to minimize activity loss. Avoid repeated freeze–thaw cycles; prepare single-use aliquots upon first thaw.
• Reconstitution (if supplied as a powder): Dissolve in a cold, compatible buffer (e.g., 50 mM sodium acetate, pH ~5.0) to the desired activity concentration. Gentle mixing; avoid vortexing to prevent foaming/denaturation.
• Storage formulations: Many enzymes are stabilized by 10–50% v/v glycerol at −20°C (remains liquid) or by freezing at −80°C without glycerol. Addition of inert carrier protein (e.g., BSA) and sugars (e.g., trehalose) can enhance stability, but only if compatible with your application. Do not add stabilizers that conflict with downstream analyses.
• Containers: Use low-protein-binding tubes; pre-chill prior to aliquoting. Rinse tips with buffer containing small amounts of a benign additive (e.g., 0.01% Tween-20) if adsorption is problematic and compatible with assays.
• Stability monitoring: Periodically verify activity using a standard sucrose assay to track any loss over storage time.

Note: Any stabilizers, preservatives, concentration, or specific activity for this catalog item are Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

Brief description: β-Fructosidase (commonly called invertase; EC 3.2.1.26) is an endo/exo glycoside hydrolase that cleaves β-D-fructofuranosidic linkages (e.g., in sucrose, raffinose, and fructans), generating fructose (and glucose from sucrose). As a protein enzyme, it does not have a single small-molecule structure.

Item-specific identifiers (from Product Data):

• SKU: F1444988
• Product Name: β-Fructosidase
• CAS: 37288-56-5
• Storage: Store at -20°C
• Shipped In: Ice chest + Ice pads

Small-molecule identifiers for this item:

• 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 protein enzymes; not specified for this item.
• InChIKey: Not applicable to protein enzymes; not specified for this item.

Literature identity and structural features (general):

• Enzyme Commission: EC 3.2.1.26 (β-fructofuranosidase)
• Enzyme families: Commonly classified within Glycoside Hydrolase families GH32 or GH68 (literature; family depends on source organism).
• Active-site features: Catalytic acid/base and nucleophile typically as conserved carboxylates (e.g., Asp/Glu) mediating a retaining mechanism via a fructofuranosyl-enzyme intermediate (literature).
• Substrate scope: β-D-fructofuranosides including sucrose (to glucose + fructose), raffinose (to melibiose + fructose), and various fructans; many sources also exhibit transfructosylation activity producing fructooligosaccharides (literature).

2D/architectural description (general):

• Typical GH32 enzymes adopt a five-bladed β-propeller catalytic domain with an associated β-sandwich/sucrose-binding subdomain; the active site accommodates the fructofuranose ring and leaving group pocket (literature).

Synthetic Utility

β-Fructosidase provides chemo-, regio-, and stereoselective transformations on β-D-fructofuranoside motifs, enabling operations that are difficult under purely chemical conditions.

Key transformations (literature):

• Selective hydrolysis: Cleavage of sucrose and related β-fructofuranosides in complex matrices without affecting α-linkages or other protecting groups, allowing stepwise deconvolution of oligosaccharides.
• Transfructosylation (retaining GH mechanism): At high sucrose concentration, transfer of fructosyl groups to acceptor hydroxyls (water-poor conditions) forms FOS with controlled DP and linkage patterns (e.g., β-(2→1) fructans).
• Late-stage editing: Enzymatic trimming of fructan chains for structural elucidation or for tuning physicochemical properties (solubility, viscosity) of carbohydrate-based materials.
• Orthogonal selectivity: Complementary to α-glucosidase and β-galactosidase, enabling orthogonal deprotection or remodeling in glycoconjugate synthesis.

Process considerations:

• Immobilized formats improve operational stability, solvent tolerance (limited), and reusability; suitable supports include epoxy-activated resins, chitosan, silica, or magnetic nanoparticles.
• Reaction engineering: Water activity, temperature, and substrate concentration strongly bias hydrolysis vs transfer. Product inhibition by glucose/fructose can be mitigated via in situ removal (e.g., yeast fermentation of glucose, membrane separation).

Limitations:

• Sensitive to heavy metals and strong oxidants; narrow optimal pH/temperature window depends on source. Organic cosolvent tolerance is generally limited; screen conditions for each substrate.

Note: Specific activity and formulation details for this catalog item are Not specified for this item; refer to CoA/Spec Sheet.

Target Specificity

This section is primarily applicable to antibodies, probes, or affinity reagents. For this enzyme product, no target-binding specificity (antigen, clone, isotype, species reactivity) is applicable.

Item-specific note: Not specified for this item; refer to CoA/Spec Sheet if any additional characterization (e.g., substrate spectrum profiling) is provided.