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Send EmailPullulanase, Debranching Enzyme, 9075-68-7
Pullulanase / Debranching Enzyme / α-Dextrin Endo-1,6-α-Glucosidase
CAS Number: 9075-68-7
EC Number: 3.2.1.41 (Enzyme Commission Number)
| Parameter | Information |
|---|---|
| Product Name | Pullulanase |
| CAS Number | 9075-68-7 |
| EC Number | 3.2.1.41 |
| Chemical Class | Enzyme (debranching enzyme, hydrolase) |
| Systematic Name | α-Dextrin Endo-1,6-α-Glucosidase |
| Source | Microbial fermentation (Bacillus acidopullulyticus, Klebsiella pneumoniae, or other microorganisms) |
| Appearance | Light yellow to white powder or liquid enzyme preparation |
| Odor | Slight characteristic odor |
| Physical State (20°C) | Solid (powder) or liquid |
Pullulanase is a debranching enzyme that specifically hydrolyzes the α-1,6-glycosidic bonds in starch, amylopectin, pullulan, and related polysaccharides. It is widely used in the starch processing industry to improve the yield of glucose and maltose syrups by breaking down branched starch structures. It is also used in the brewing and beverage industries for sugar profile adjustment.
| Type | Name |
|---|---|
| Common Names | Pullulanase, Debranching Enzyme |
| Systematic Name | α-Dextrin Endo-1,6-α-Glucosidase |
| EC Number | 3.2.1.41 |
| CAS Number | 9075-68-7 |
| HS Code | 3507.90 |
Pullulanase is a hydrolytic enzyme that specifically cleaves α-1,6-glycosidic bonds. It is a single polypeptide chain with a molecular weight of approximately 100,000 Da (100 kDa), depending on the microbial source.
Structural Features:
N-Terminus
|
[Catalytic Domain]
|
[Starch Binding Domain]
|
C-Terminus
| Feature | Description |
|---|---|
| Molecular Weight | ~100,000 Da (100 kDa) |
| Structure | Single polypeptide chain |
| Active Site | Hydrolytic domain (cleaves α-1,6 bonds) |
| Optimal pH | 4.5 – 6.0 |
| Optimal Temperature | 50 – 60°C |
Pullulanase catalyzes the hydrolysis of α-1,6-glycosidic bonds in amylopectin, pullulan, and other branched polysaccharides, releasing linear chains.
Reaction:
Pullulan + H₂O → Maltotriose + Maltose + Glucose
(α-1,6 bonds cleaved)
General Reaction:
Amylopectin (α-1,6 bonds) + H₂O → Amylopectin (linear chains) + Oligosaccharides
| Property | Value |
|---|---|
| Appearance | Light yellow to white powder or liquid |
| Odor | Slight characteristic odor |
| Solubility | Soluble in water |
| Molecular Weight | ~100,000 Da (100 kDa) |
| Optimal pH Range | 4.5 – 6.0 |
| Effective pH Range | 4.0 – 7.0 |
| Optimal Temperature | 50 – 60°C |
| Effective Temperature Range | 30 – 70°C |
| Inactivation Temperature | >70°C |
| Calcium Dependency | Calcium-independent (some variants) |
| Moisture Content (Powder) | ≤ 8% |
| Ash Content | ≤ 2% |
| Heavy Metals | ≤ 10 ppm |
| Parameter | Value |
|---|---|
| Activity Definition | One unit (U) is defined as the amount of enzyme that liberates 1 µmol of reducing sugar (as glucose) per minute at pH 5.0 and 50°C using pullulan as substrate. |
| Typical Activity | Varies by manufacturer (typically 100-500 U/g or U/mL) |
| Substrate | Pullulan, amylopectin, glycogen, α-1,6-glycosidic bonds |
| Reaction Product | Maltotriose, maltose, glucose, and linear oligosaccharides |
| Step | Description |
|---|---|
| 1 | Pullulanase binds to branched regions of amylopectin, pullulan, or glycogen. |
| 2 | The enzyme recognizes and binds to α-1,6-glycosidic bonds at branch points. |
| 3 | Hydrolysis of the α-1,6-glycosidic bond occurs. |
| 4 | The branched structure is cleaved, releasing linear chains. |
| 5 | The enzyme continues to hydrolyze remaining α-1,6 bonds until the substrate is fully debranched. |
| Application | Function |
|---|---|
| Starch Hydrolysis | Breaks down branched starch structures (amylopectin) to increase glucose yield |
| High-DE Syrups | Improves dextrose equivalent (DE) values in glucose syrup production |
| Saccharification | Used in combination with amyloglucosidase for complete starch conversion |
| Yield Improvement | Increases glucose yield by 2-5% compared to using amyloglucosidase alone |
| Application | Function |
|---|---|
| High-Maltose Syrups | Produces high-maltose syrups for confectionery and brewing |
| Sugar Profile Adjustment | Controls the ratio of maltose to glucose in the syrup |
| Application | Function |
|---|---|
| Beer Production | Sugar profile adjustment; improves fermentability |
| Light Beer | Reduces carbohydrate content for light beer production |
| Low-Carbohydrate Beer | Increases fermentable sugar content for low-carb beers |
| Application | Function |
|---|---|
| High-Fructose Corn Syrup (HFCS) | Used in combination with amyloglucosidase and glucose isomerase |
| Sugar Alcohols | Used in the production of maltitol, sorbitol, and other sugar alcohols |
| Application | Function |
|---|---|
| Bioethanol Production | Improves starch conversion for ethanol fermentation |
| Animal Feed | Improves digestibility of starch in animal feed |
| Textile Industry | Desizing of starch-based sizing agents |
| Paper Industry | Starch modification |
| Application | Recommended Dosage | Conditions |
|---|---|---|
| Glucose Syrup Production | 0.1 – 0.5 U/g starch | pH 4.5-5.5, 55-60°C |
| Maltose Syrup Production | 0.2 – 1.0 U/g starch | pH 5.0-6.0, 50-55°C |
| Brewing (Light Beer) | 0.05 – 0.2 U/g malt | pH 4.5-5.5, 50-55°C |
| High-Fructose Corn Syrup | 0.1 – 0.3 U/g starch | pH 4.5-5.5, 55-60°C |
| Bioethanol Production | 0.1 – 0.5 U/g starch | pH 4.5-5.5, 50-55°C |
Note: Dosage should be optimized for each specific application through trial runs.
| Parameter | Recommendation |
|---|---|
| Dissolution | Dissolve in water or buffer (pH 4.5-6.0) at room temperature |
| Optimal Temperature | 50-60°C |
| Optimal pH | 4.5-6.0 |
| Reaction Time | 2-24 hours (depending on substrate and enzyme concentration) |
| Inactivation | Heat treatment (>70°C) or pH adjustment to inactivate the enzyme |
| Substrate Concentration | 20-40% w/w (starch dry solids) |
| Factor | Effect |
|---|---|
| Temperature | Optimal at 50-60°C; activity decreases above 65°C; >70°C inactivation |
| pH | Optimal at pH 4.5-6.0; significant activity at pH 4.0-7.0 |
| Substrate Concentration | Activity increases with substrate concentration up to saturation |
| Calcium Ions | Most pullulanases are calcium-independent |
| Inhibitors | Heavy metals (Cu²⁺, Hg²⁺, Pb²⁺) may inhibit activity |
| Competing Enzymes | May be used in combination with amyloglucosidase and α-amylase |
| Parameter | Value |
|---|---|
| Acute Oral Toxicity (LD50, Rat) | > 5,000 mg/kg (very low toxicity) |
| Acute Dermal Toxicity | Low |
| Acute Inhalation Toxicity | May cause respiratory sensitization (dust) |
| Skin Irritation | Mild irritant |
| Eye Irritation | May cause mild irritation |
| Skin Sensitization | May cause sensitization in sensitive individuals |
| Mutagenicity | Negative (Ames test) |
| Carcinogenicity | Not classified as a carcinogen |
| Reproductive Toxicity | Not classified as a reproductive toxin |
| Allergenicity | May cause allergic reactions in sensitive individuals (protein-based) |
| Hazard Class | Category | H-Statement |
|---|---|---|
| Respiratory Sensitization | Category 1 | H334 |
| Skin Sensitization | Category 1 | H317 |
Signal Word: WARNING
Hazard Pictograms: GHS07 (Exclamation Mark), GHS08 (Health)
Hazard Statements (H-Codes):
| Code | Statement |
|---|---|
| H317 | May cause an allergic skin reaction. |
| H334 | May cause allergy or asthma symptoms or breathing difficulties if inhaled. |
| Code | Statement |
|---|---|
| P261 | Avoid breathing dust/fume/gas/mist/vapors/spray. |
| P264 | Wash hands thoroughly after handling. |
| P271 | Use only outdoors or in a well-ventilated area. |
| P280 | Wear protective gloves/protective clothing/eye protection/face protection. |
| P302+P352 | IF ON SKIN: Wash with plenty of soap and water. |
| P304+P340 | IF INHALED: Remove person to fresh air and keep comfortable for breathing. |
| P305+P351+P338 | IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses if present. |
| P342+P311 | If experiencing respiratory symptoms: Call a POISON CENTER/doctor. |
| P333+P313 | If skin irritation or rash occurs: Get medical advice/attention. |
| P337+P313 | If eye irritation persists: Get medical advice/attention. |
| P362+P364 | Take off contaminated clothing and wash it before reuse. |
| P501 | Dispose of contents/container in accordance with local regulations. |
| Route of Exposure | Action to Be Taken |
|---|---|
| Inhalation | Move to fresh air. If respiratory irritation persists, seek medical attention. |
| Skin Contact | Wash with plenty of soap and water. Remove contaminated clothing. If irritation persists, consult a physician. |
| Eye Contact | Rinse thoroughly with plenty of water for at least 15 minutes. Remove contact lenses. If irritation persists, seek medical attention. |
| Ingestion | Rinse mouth with water. Drink plenty of water. Do not induce vomiting. If large amount is swallowed, seek medical attention. |
| Parameter | Information |
|---|---|
| Fire Hazard | Combustible organic powder; may burn at high temperatures. |
| Hazards During Fire | Thermal decomposition produces toxic fumes including carbon monoxide, carbon dioxide, and nitrogen oxides. |
| Suitable Extinguishing Media | Water spray, CO₂, dry chemical powder, alcohol-resistant foam. |
| Special Protective Equipment | Self-contained breathing apparatus (SCBA), full protective clothing. |
| Parameter | Information |
|---|---|
| Personal Protection | Dust mask, protective goggles, gloves, coverall. |
| Ventilation | Increase ventilation; use local exhaust in confined spaces. |
| Containment | Cover spilled area to prevent dust dispersion. |
| Cleaning Methods | Collect dry using vacuum or careful sweeping. Avoid washing with water (soluble). |
| Environmental Precautions | Prevent entry into drains, sewers, and water bodies. |
| Waste Disposal | Dispose of in accordance with local regulations. |
| Parameter | Information |
|---|---|
| Storage Conditions | Store in a cool, dry, and well-ventilated area at 4-25°C. |
| Container Requirements | Keep tightly closed in original packaging. Protect from moisture and direct sunlight. |
| Materials to Avoid | Strong oxidizers, high heat, moisture. |
| Shelf Life | 12-24 months (in unopened original packaging under proper storage). |
| Stability Note | Enzyme activity decreases over time; store at recommended temperatures to maintain activity. |
| Packaging Options | Aluminum foil bags, plastic containers. |
| Packaging Type | Quantity | Material |
|---|---|---|
| Aluminum Foil Bag | 1 kg | Laminated aluminum |
| Plastic Container | 1 kg, 5 kg, 10 kg | HDPE / Food-grade plastic |
| Fiber Drum | 25 kg | Cardboard / Plastic |
| Parameter | Information |
|---|---|
| UN Number | Not applicable (not classified as dangerous goods) |
| Hazard Class | Not classified as hazardous for transport |
| Packing Group | Not applicable |
| ADR/RID | Not regulated |
| IMDG Code | Not regulated |
| IATA (Air) | Not regulated |
| Marine Pollutant | No |
| Transport Temperature | Ambient temperature; protect from moisture. |
| Region / Authority | Status |
|---|---|
| European Union | Approved as a food enzyme (EU 1332/2008). |
| USA (FDA) | GRAS (Generally Recognized as Safe) for food applications. |
| Turkey | Approved as a food processing aid. |
| JECFA | Approved as a food enzyme. |
| EFSA | Re-evaluated and considered safe at current usage levels. |
| Kosher/Halal | Can be certified upon request. |
| Parameter | Information |
|---|---|
| Aquatic Toxicity | Low toxicity to aquatic organisms. |
| Biodegradability | Readily biodegradable (protein-based). |
| Bioaccumulation | Not expected. |
| Waste Disposal | Dispose of in accordance with local regulations. Incineration is recommended for contaminated materials. |
| Property | Pullulanase | Amyloglucosidase | α-Amylase | Isoamylase |
|---|---|---|---|---|
| Bond Cleaved | α-1,6 | α-1,4 | α-1,4 | α-1,6 |
| Substrate | Pullulan, amylopectin | Starch, dextrins | Starch, dextrins | Amylopectin |
| Optimal pH | 4.5-6.0 | 4.0-5.0 | 5.5-6.5 | 3.5-5.0 |
| Optimal Temperature | 50-60°C | 55-65°C | 70-90°C | 45-55°C |
| Product | Linear oligosaccharides | Glucose | Dextrins | Linear chains |
| Primary Use | Debranching | Saccharification | Liquefaction | Debranching |
Q1: What is pullulanase used for?
A: Pullulanase is used to hydrolyze α-1,6-glycosidic bonds (branch points) in starch, amylopectin, and pullulan. It is primarily used in the production of glucose syrups, maltose syrups, high-fructose corn syrup, and in the brewing industry.
Q2: Is pullulanase safe to eat?
A: Yes. Pullulanase is considered safe (GRAS by FDA, approved by EFSA and JECFA) for food applications. The enzyme is inactivated during processing and is not present in the final product.
Q3: What is the difference between pullulanase and isoamylase?
A: Both are debranching enzymes that hydrolyze α-1,6-glycosidic bonds. Pullulanase is more active on pullulan and is typically more thermostable. Isoamylase is more active on glycogen and amylopectin, with a lower optimal pH.
Q4: Can pullulanase be used with other enzymes?
A: Yes. Pullulanase is often used in combination with amyloglucosidase (for complete saccharification) and α-amylase (for liquefaction) to achieve high glucose yields.
Q5: What is the optimal temperature and pH for pullulanase?
A: The optimal temperature is 50-60°C, and the optimal pH is 4.5-6.0. The enzyme is inactivated above 70°C.
Q6: How should pullulanase be stored?
A: Store in a cool, dry place at 4-25°C, in tightly closed original packaging. Protect from moisture and direct sunlight.
| Property | Value |
|---|---|
| CAS Number | 9075-68-7 |
| EC Number | 3.2.1.41 |
| Molecular Weight | ~100,000 Da (100 kDa) |
| Appearance | Light yellow to white powder or liquid |
| Solubility | Soluble in water |
| Optimal pH | 4.5 – 6.0 |
| Optimal Temperature | 50 – 60°C |
| Inactivation Temperature | >70°C |
| Typical Activity | 100-500 U/g or U/mL |
| Primary Uses | Glucose syrup production, brewing, sweetener production |
| Shelf Life | 12-24 months |
| UN Number | Not applicable |
| GHS Signal Word | WARNING |
CRITICAL NOTICES:
Respiratory Sensitization: Pullulanase powder may cause respiratory sensitization (H334) in susceptible individuals. Use appropriate dust control measures and respiratory protection.
Skin Sensitization: May cause allergic skin reactions (H317) in sensitive individuals. Use protective gloves.
Enzyme Activity: Enzyme activity decreases with temperature, pH, and time. Optimize conditions for each application to achieve desired results.
Heat Inactivation: The enzyme is inactivated above 70°C. This ensures that no active enzyme remains in the final product after processing.
Calcium Dependency: Most microbial pullulanases are calcium-independent, simplifying process conditions compared to calcium-dependent enzymes.
Dosage Optimization: Recommended dosages are starting points. Conduct trial runs to optimize the dosage for specific applications and products.
Substrate Specificity: Pullulanase is highly specific for α-1,6-glycosidic bonds. The presence of branched structures is essential for activity.
Storage: Store at recommended temperatures (4-25°C). Higher temperatures accelerate activity loss.
BEST PRACTICE RECOMMENDATIONS:
Storage: Store in a cool, dry, and well-ventilated area at 4-25°C. Keep containers tightly closed. Protect from moisture and direct sunlight.
Handling: Use dust mask, protective gloves, and goggles. Avoid dust formation. Ensure adequate ventilation.
Dissolution: Dissolve in water or buffer (pH 4.5-6.0) at room temperature. Use immediately after dissolution.
Application: Add the enzyme to the starch solution at the appropriate stage of processing. Ensure uniform distribution.
Incubation: Allow sufficient reaction time at the optimal temperature for the enzyme to act. Reaction time varies by application.
Inactivation: Ensure the final product is heated to >70°C or pH-adjusted to inactivate the enzyme.
Combination: For maximum glucose yield, combine pullulanase with amyloglucosidase during saccharification.
Dosage: Start with recommended dosages and optimize through trial runs. Overdosing may cause excessive hydrolysis and undesirable sugar profiles.
Quality Control: Monitor enzyme activity and product quality regularly. Test for residual enzyme activity if required.
Waste Management: Dispose of waste in accordance with local regulations.
LEGAL DISCLAIMER:
This Technical Data Sheet (TDS) is for informational purposes only and is prepared based on available technical data. The user is solely responsible for determining the suitability of the product for their specific application and for complying with all local, national, and international regulations. For complete safety, storage, handling, transport, waste, and regulatory compliance information, the official Safety Data Sheet (SDS/MSDS) provided by the manufacturer/supplier must be consulted. This document does not substitute professional advice. Dosage and application conditions should be optimized through product-specific trials.