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Alpha Amylase, Bacterial Alpha-Amylase, 9000-90-2

Alpha Amylase, Bacterial Alpha-Amylase, 9000-90-2

ALPHA-AMYLASE ENZYMES

1. PRODUCT DEFINITION

Alpha-Amylase (EC 3.2.1.1) is a hydrolase enzyme that hydrolyzes α-1,4-glycosidic bonds in polysaccharides such as starch and glycogen, producing smaller sugars (maltose, glucose, dextrins, oligosaccharides). In the food industry, different types derived from various sources (fungal, bacterial, plant-based) are used depending on process conditions. The enzyme acts by randomly cleaving internal bonds (endo-effect) within the starch chain, rapidly reducing viscosity and achieving liquefaction.

2. CAS NUMBER CLARIFICATION AND VERIFICATION

There is widespread confusion regarding CAS numbers for alpha-amylase enzymes. The following table clarifies the correct CAS numbers:

CAS Number Enzyme Type Source Description
9001-19-8 Fungal Alpha-Amylase Aspergillus oryzae CORRECT: Official CAS number for fungal (mold) derived alpha-amylase. Suitable for baking and food applications.
9000-90-2 Bacterial Alpha-Amylase Bacillus species CORRECT: Official CAS number for bacterial derived alpha-amylase. Suitable for industrial applications.
9000-85-5 Alternative / Old Designation - Appears in some old catalogs as an alternative or general alpha-amylase designation. Not valid in current official classifications.

CRITICAL NOTICE:

  • CORRECT CAS number for Fungal Alpha-Amylase: 9001-19-8

  • CORRECT CAS number for Bacterial Alpha-Amylase: 9000-90-2

  • 9000-85-5 is an old/alternative designation and is not valid in current official classifications.

  • Always verify the correct CAS number on the supplier's Safety Data Sheet (SDS).

3. PRODUCT TYPES AND SOURCES

Type Source CAS No EC No Optimal pH Optimal Temperature Application Area
Fungal Alpha-Amylase Aspergillus oryzae 9001-19-8 3.2.1.1 4.5 – 6.0 50 – 60°C Baking, cakes, biscuits, confectionery, dough conditioning
Bacterial Alpha-Amylase Bacillus amyloliquefaciens 9000-90-2 3.2.1.1 6.0 – 7.0 60 – 75°C Brewing, starch sugars, modified starch, detergents
High Temperature Bacterial Alpha-Amylase Bacillus licheniformisGeobacillus stearothermophilus 9000-90-2 3.2.1.1 6.0 – 7.0 70 – 90°C High-temperature starch hydrolysis, HFCS, bioethanol
Plant-Based Alpha-Amylase Barley, wheat, rice 9000-90-2 3.2.1.1 5.5 – 6.5 30 – 40°C Malt, beer, whisky, traditional fermented beverages

4. CHEMICAL STRUCTURE AND MOLECULAR PROPERTIES

4.1. Enzyme Classification

Feature Description
Enzyme Class Hydrolase
Subclass Glycosidase
Systematic Name 1,4-alpha-D-glucan glucanohydrolase
Reaction Endohydrolysis of 1,4-alpha-D-glycosidic bonds
CAZy Family Glycoside Hydrolase Family 13 (GH13)
Structural Fold (β/α)₈ TIM Barrel

4.2. Molecular Structure

Alpha-Amylases belong to Glycoside Hydrolase Family 13 (GH13) and adopt a characteristic (β/α)₈ TIM Barrel structure. This structure consists of 8 parallel β-strands surrounded by 8 α-helices forming a barrel shape. The active site is located at the C-terminal end of the barrel and contains conserved aspartate and glutamate residues. Calcium ions are essential for structural stability and thermal stability of the enzyme.

Domain Architecture:

                    ALPHA-AMYLASE
                    (GH13 Family - TIM Barrel)
                           |
        +------------------+------------------+
        |                  |                  |
    Domain A         Domain B         Domain C
    (Catalytic)    (Loop/Calcium)   (C-terminal)
    (β/α)₈ TIM        Ca²⁺             β-sheet
    Barrel           Binding          Structure

Domain Description:

Domain Location Function
Domain A (Catalytic) Central core (β/α)₈ barrel Contains catalytic site; Responsible for substrate binding and hydrolysis
Domain B Insertion between β3-α3 loops Calcium binding site; Contributes to structural stability
Domain C C-terminal region β-sheet rich region; May be involved in substrate recognition and stability

4.3. Active Site Architecture

The active site of alpha-amylase contains conserved amino acid residues essential for catalytic activity:

              CATALYTIC RESIDUES
                    |
        +-----------+-----------+
        |           |           |
      Asp-X-Asp     Glu       Asp
    (Nucleophile) (Acid/Base) (Transition)
Residue Function Location
Asp (Aspartate) Nucleophile; Attacks the glycosidic bond Domain A (β4 strand)
Glu (Glutamate) Acid/Base catalyst; Protonates the leaving group Domain A (β4 strand)
Asp (Aspartate) Stabilizes the transition state Domain A (β5 strand)

4.4. Calcium Binding Site

Calcium ions (Ca²⁺) are essential for structural stability and catalytic activity of alpha-amylase:

              CALCIUM BINDING SITE
                    |
        +-----------+-----------+
        |           |           |
       Asp        Asp          Asp
       (Oxygen)   (Oxygen)    (Oxygen)
                    |
                   Ca²⁺
                    |
       Asp        Asn          His
       (Oxygen)   (Oxygen)    (Oxygen)
Feature Description
Calcium Ion Binds near the active site with high affinity
Coordination 7-8 coordination bonds with oxygen atoms from Asp, Asn, His residues
Function Maintains structural integrity; Protects against thermal denaturation; Essential for optimal activity
Replacement Other divalent cations (Mg²⁺, Mn²⁺) can partially substitute but with reduced stability

4.5. Structural Visualization

(β/α)₈ TIM Barrel Schematic Representation:

                     TOP VIEW
                 
                       α8
                  ____/____
                 /    α1    \
                /  ________  \
               |  / α2    \  |
               | |  ______  | |
               | | / α3   \ | |
               | | |  β4  | | |
               | | | /β5\ | | |
               | | | |   | | | |
               | | | \β6/ | | |
               | | |  β7  | | |
               | | \ α8  / | |
               | |  β8   | | |
               | | \____/ | | |
               | |  α9   | | |
               |  \ α10 /  | |
                \  α11  /  /
                 \____/ α12
                    |
              Catalytic Site
           (Asp, Glu, Asp residues)

Substrate Binding and Catalysis Scheme:

                   SUBSTRATE (STARCH)
                          |
                   α-1,4 Glycosidic Bond
                          |
            -O---[Glucose]---O---[Glucose]---O-
                      |                 |
                   Cleavage Site
                          |
                 CATALYTIC RESIDUES
                          |
          Asp (Nucleophile)  Glu (Acid/Base)
                          |
                 HYDROLYSIS PRODUCTS
                          |
            Oligosaccharides + Maltose + Glucose

4.6. Molecular Properties by Type

Property Fungal Bacterial High Temp. Plant-Based
Molecular Weight ~50-60 kDa ~50-70 kDa ~50-70 kDa ~45-55 kDa
Amino Acid Count ~450-500 ~500-520 ~500-520 ~400-480
Isoelectric Point (pI) ~4.0 – 5.0 ~4.5 – 5.5 ~4.5 – 5.5 ~5.5 – 6.5
Calcium Binding 1 Ca²⁺ 2 Ca²⁺ 3-4 Ca²⁺ 1 Ca²⁺
Disulfide Bonds 1-2 2-3 3-4 1-2
Glycosylation None None None None
Optimal pH 4.5 – 6.0 6.0 – 7.0 6.0 – 7.0 5.5 – 6.5
Optimal Temperature 50 – 60°C 60 – 75°C 70 – 90°C 30 – 40°C
Thermostability Low-Moderate High Very High Low

5. MECHANISM OF ACTION (DETAILED)

5.1. Catalytic Cycle

Step 1: Substrate Binding
The starch molecule binds to the enzyme's active site through hydrogen bonds and hydrophobic interactions. The substrate binds in a deep cleft at the C-terminal end of the TIM barrel. Conformational changes occur in the enzyme upon binding.

Step 2: Nucleophilic Attack
The catalytic aspartate residue acts as a nucleophile, attacking the anomeric carbon of the glycosidic bond. This forms a covalent glycosyl-enzyme intermediate.

Step 3: Acid/Base Catalysis
The catalytic glutamate residue acts as an acid/base catalyst. It donates a proton to the glycosidic oxygen (leaving group), facilitating bond cleavage.

Step 4: Transition State Stabilization
A second aspartate residue stabilizes the developing positive charge in the transition state, lowering the activation energy of the reaction.

Step 5: Product Release
The glycosyl-enzyme intermediate is hydrolyzed by water, releasing the product (oligosaccharide). The enzyme returns to its ground state, ready for another catalytic cycle.

                CATALYTIC CYCLE

    Enzyme + Starch ──────────────→ Enzyme-Starch Complex
                   (Substrate Binding)
    
    Enzyme-Starch Complex ────────→ Glycosyl-Enzyme + Leaving Group
                   (Nucleophilic Attack / Acid Catalysis)
    
    Glycosyl-Enzyme + Water ─────→ Enzyme + Hydrolysis Product
                   (Hydrolysis / Product Release)

5.2. Mode of Action (Endo vs Exo)

Feature Alpha-Amylase (Endo) Beta-Amylase (Exo) Glucoamylase (Exo)
Cleavage Mode Random internal cleavage Removes maltose from termini Removes glucose from termini
Viscosity Reduction Rapid Slow Slow
Products Dextrins, maltose, glucose Maltose Glucose
Liquefaction Rapid Slow Slow

6. PHYSICAL AND CHEMICAL PROPERTIES (DETAILED)

6.1. General Properties

Property Fungal Bacterial High Temp. Plant-Based
Appearance Light brown powder/liquid Light brown powder/liquid Light brown powder/liquid Light brown powder/liquid
Odor Slight enzyme odor Slight enzyme odor Slight enzyme odor Slight enzyme odor
Solubility Soluble in water Soluble in water Soluble in water Soluble in water
Optimal pH 4.5 – 6.0 6.0 – 7.0 6.0 – 7.0 5.5 – 6.5
Effective pH Range 4.0 – 8.0 5.0 – 9.0 5.0 – 8.5 4.5 – 7.5
Optimal Temperature 50 – 60°C 60 – 75°C 70 – 90°C 30 – 40°C
Effective Temperature Range 30 – 70°C 40 – 85°C 50 – 105°C 20 – 55°C
Inactivation Temperature >65-70°C >85-90°C >100-105°C >55-60°C
Thermostability Low-Moderate High Very High Low
Calcium Requirement Moderate High High Low
Isoelectric Point (pI) ~4.0 – 5.0 ~4.5 – 5.5 ~4.5 – 5.5 ~5.5 – 6.5
Molecular Weight ~50-60 kDa ~50-70 kDa ~50-70 kDa ~45-55 kDa

6.2. Activity Properties

Property Fungal Bacterial High Temp. Plant-Based
Specific Activity 10,000-20,000 U/g 10,000-30,000 U/g 10,000-30,000 U/g 5,000-15,000 U/g
Km (Starch) ~1-2 mg/mL ~2-5 mg/mL ~2-4 mg/mL ~1-3 mg/mL
Kcat ~500 s⁻¹ ~800 s⁻¹ ~1000 s⁻¹ ~400 s⁻¹
pH Stability 4.0 – 8.0 5.0 – 9.0 5.0 – 8.5 4.5 – 7.5
Thermal Stability Up to 70°C Up to 90°C Up to 105°C Up to 55°C
Calcium Effect 1.5-2x increase 2-3x increase 2-3x increase 1.2x increase

6.3. Activity Unit Definition

International Unit (U): 1 U is defined as the amount of enzyme that liberates 1 µmol of reducing sugar (as glucose equivalent) per minute at pH 6.0 and 40°C using liquefied starch as substrate. Measurement conditions and substrates may vary by type.

7. COMPARISON BETWEEN TYPES (20+ POINTS)

7.1. Source and Production Differences

  1. Fungal Alpha-Amylase: Derived from Aspergillus oryzae or A. niger. Produced by solid-state or submerged fermentation. Requires 24-48 hours fermentation time.

  2. Bacterial Alpha-Amylase: Derived from Bacillus amyloliquefaciens or B. subtilis. Produced by submerged fermentation. Faster production with 12-24 hours fermentation time.

  3. High Temperature Alpha-Amylase: Derived from Bacillus licheniformis or Geobacillus stearothermophilus. Requires thermophilic production conditions (55-65°C). 12-18 hours fermentation time.

  4. Plant-Based Alpha-Amylase: Extracted from cereals such as barley, wheat, rice, rye, oats. Requires germination (malting) process. 5-7 days germination time.

7.2. Physical and Chemical Differences

  1. Fungal Alpha-Amylase: Active at pH 4.5-6.0. Ideal for baking and acidic food processes.

  2. Bacterial Alpha-Amylase: Active at pH 6.0-7.0. Suitable for neutral and slightly alkaline industrial processes.

  3. High Temperature Alpha-Amylase: Active at pH 6.0-7.0. Remains active at 90-105°C, critical for starch liquefaction.

  4. Plant-Based Alpha-Amylase: Active at pH 5.5-6.5. Works at low temperatures (30-40°C), suitable for malt and beer production.

  5. Calcium Requirement: Fungal → Moderate (1 mM Ca²⁺); Bacterial and High Temp. → High (2-5 mM Ca²⁺); Plant-Based → Low (0.5 mM Ca²⁺).

  6. Inactivation Temperature: Fungal >65-70°C; Bacterial >85-90°C; High Temp. >100-105°C; Plant-Based >55-60°C.

  7. Molecular Weight: Fungal ~50-60 kDa; Bacterial ~50-70 kDa; High Temp. ~50-70 kDa; Plant-Based ~45-55 kDa.

  8. Isoelectric Point: Fungal 4.0-5.0; Bacterial 4.5-5.5; High Temp. 4.5-5.5; Plant-Based 5.5-6.5.

  9. Disulfide Bonds: Fungal 1-2; Bacterial 2-3; High Temp. 3-4; Plant-Based 1-2. More disulfide bonds provide higher thermal stability.

  10. Glycosylation: None in all types (prokaryotic and fungal enzymes are not glycosylated).

7.3. Substrate and Product Differences

  1. Substrate Preference: Fungal → Amylose > Amylopectin; Bacterial → Amylopectin > Amylose; High Temp. → Amylopectin > Amylose; Plant-Based → Amylose ≈ Amylopectin.

  2. Product Profile: Fungal → Dextrins + Maltose + Glucose (5-15%); Bacterial → Dextrins + Maltose (15-25%); High Temp. → Dextrins + Maltose (15-25%); Plant-Based → Maltose (60-80%) + Dextrins.

  3. Hydrolysis Rate: High Temp. > Bacterial > Fungal > Plant-Based (depending on temperature).

  4. Substrate Affinity (Km): Fungal ~1-2 mg/mL (high affinity); Bacterial ~2-5 mg/mL; High Temp. ~2-4 mg/mL; Plant-Based ~1-3 mg/mL.

7.4. Industrial Application Differences

  1. Main Application Area: Fungal → Baking, cakes, biscuits; Bacterial → Starch sugars, modified starch; High Temp. → HFCS, bioethanol; Plant-Based → Malt, beer, whisky.

  2. Process Conditions: Fungal → 50-60°C, pH 4.5-6.0; Bacterial → 60-75°C, pH 6.0-7.0; High Temp. → 70-90°C, pH 6.0-7.0; Plant-Based → 30-40°C, pH 5.5-6.5.

  3. Typical Dosage: Fungal → 2 g per 50 kg flour; Bacterial → 0.5-2.0 U/g starch; High Temp. → 0.5-2.0 U/g starch; Plant-Based → Varies with malt quantity.

  4. Regulatory Status: Fungal → FDA GRAS, EFSA approved (food); Bacterial → FDA GRAS, EFSA approved (industrial); High Temp. → FDA GRAS, EFSA approved (industrial); Plant-Based → Traditional use, GRAS.

  5. Shelf Life: Fungal → 12-24 months (4-25°C); Bacterial → 12-24 months (4-25°C); High Temp. → 12-24 months (4-25°C); Plant-Based → 6-12 months (4-25°C, more sensitive).

  6. Cost/Performance: Fungal → Moderate cost, high performance (food); Bacterial → Low cost, moderate-high performance; High Temp. → High cost, very high performance; Plant-Based → Low cost, low-moderate performance.

8. APPLICATION AREAS (DETAILED)

8.1. Baking Industry (Fungal Alpha-Amylase - CAS 9001-19-8)

Application Function Mechanism
Bread Making Breaks down starch in flour; releases fermentable sugars; improves crust color and flavor; increases bread volume and texture Endo-amylase activity cuts starch chains; creates sugar source for yeast
Cakes and Biscuits Provides more flexible dough; improves gas retention; creates finer and more uniform crumb structure Viscosity reduction and gas retention improvement
Confectionery Modifies starch structure to retard staling; extends freshness and shelf life Reduces amylopectin retrogradation
Whole Wheat Bread Breaks down bran starch; improves volume and texture Hydrolysis of bran starch
Frozen Dough Improves freeze-thaw stability Modification of starch structure
Gluten-Free Bread Breaks down starch in gluten-free flours; improves texture and volume Modification of gluten-free starch matrix

Typical Dosage: 2 g per 50 kg flour

8.2. Brewing Industry (Bacterial/Plant-Based Alpha-Amylase - CAS 9000-90-2)

Application Function Mechanism
Mashing Converts grain starch to fermentable sugars Starch liquefaction and saccharification
Viscosity Reduction Reduces mash viscosity; improves filtration and yield Breakdown of starch chains
Malt Production Breaks down starch in barley, wheat, and rice Complements endogenous enzyme activity
Lager Beer Creates suitable sugar profile for low-temperature fermentation Controlled hydrolysis
Ale Beer Creates suitable sugar profile for high-temperature fermentation Higher-temperature hydrolysis
Gluten-Free Beer Breaks down starch in gluten-free grains Gluten-free starch hydrolysis

Typical Dosage: 0.4 – 0.8 L/ton (beer)

8.3. Starch Sugar Industry (Bacterial/High Temp. - CAS 9000-90-2)

Application Function Mechanism
Glucose Syrup Complete hydrolysis of starch to produce glucose Liquefaction (alpha-amylase), then saccharification (glucoamylase)
High Fructose Corn Syrup (HFCS) High-temperature liquefaction stage Starch hydrolysis at 90-105°C
Maltose Syrup Controlled hydrolysis for maltose production Limited hydrolysis for high maltose content
Modified Starch Adjustment of starch chain length Limited hydrolysis for specific chain length
Maltodextrin Starch hydrolysis for maltodextrin production Hydrolysis to DE 10-20
Cyclodextrin Pretreatment for cyclodextrin production Starch liquefaction

Typical Dosage: 0.5 – 2.0 U/g starch

8.4. Other Industries

Sector Application Enzyme Type
Textile Desizing (removal of starch-based sizing agents) Bacterial / High Temp.
Detergent Removal of starch-based stains Bacterial / High Temp. (thermostable)
Animal Feed Digestibility enhancement Fungal / Bacterial
Bioethanol Conversion of starch to fermentable sugars Bacterial / High Temp.
Paper Modification of starch-based coatings Bacterial
Cosmetics Modification of starch-based ingredients Fungal
Pharmaceutical Starch modification as excipient Bacterial

9. QUALITY SPECIFICATIONS

9.1. Food Grade

Parameter Fungal Bacterial High Temp. Test Method
Appearance Light brown powder Light brown powder Light brown powder Visual
Odor Slight enzyme odor Slight enzyme odor Slight enzyme odor Olfactometric
Activity 10,000 – 20,000 U/g 10,000 – 30,000 U/g 10,000 – 30,000 U/g Colorimetric
Loss on Drying ≤ 8.0% ≤ 8.0% ≤ 8.0% Gravimetric
Total Plate Count ≤ 50,000 CFU/g ≤ 50,000 CFU/g ≤ 50,000 CFU/g Microbiological
Salmonella Negative Negative Negative Microbiological
E. coli Negative Negative Negative Microbiological
Yeast and Mold ≤ 100 CFU/g ≤ 100 CFU/g ≤ 100 CFU/g Microbiological
Heavy Metals (as Pb) ≤ 10 ppm ≤ 10 ppm ≤ 10 ppm AAS / ICP
Lead (Pb) ≤ 2 ppm ≤ 2 ppm ≤ 2 ppm AAS / ICP
Arsenic (As) ≤ 2 ppm ≤ 2 ppm ≤ 2 ppm AAS / ICP
Total Protein Variable Variable Variable Kjeldahl
pH (1% Solution) 5.0 – 6.5 6.0 – 7.5 6.0 – 7.5 pH Meter

9.2. Feed Grade

Parameter Fungal Bacterial Test Method
Activity 5,000 – 10,000 U/g 5,000 – 15,000 U/g Colorimetric
Loss on Drying ≤ 10.0% ≤ 10.0% Gravimetric
Total Plate Count ≤ 100,000 CFU/g ≤ 100,000 CFU/g Microbiological
Salmonella Negative Negative Microbiological
E. coli Negative Negative Microbiological

10. TOXICOLOGICAL PROFILE

Parameter Value
Acute Oral Toxicity (LD50, Rat) > 2,000 mg/kg (low toxicity)
Acute Dermal Toxicity Low toxicity; Not an irritant
Skin Irritation Not an irritant (enzyme proteins may cause mild irritation)
Eye Irritation Mild irritant
Inhalation Toxicity May cause allergic sensitization (enzyme dust)
Chronic Toxicity Not toxic at normal usage levels
Carcinogenicity Not classified as a carcinogen
Mutagenicity Not mutagenic (Ames test negative)
Reproductive Toxicity No reproductive toxicity at normal levels
Allergenicity Enzymes are potential allergens; May cause respiratory sensitization in sensitive individuals
ADI (Acceptable Daily Intake) Not determined (enzyme is inactivated during processing)
Food Safety FDA GRAS, EFSA approved, JECFA evaluated

11. GHS CLASSIFICATION

Hazard Class Category H-Statement
Respiratory Sensitization Category 1 H334
Skin Sensitization Category 1 H317
Water Hazard (WGK Germany) 3 (severe hazard to water) -

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.

12. PRECAUTIONARY STATEMENTS (P-CODES)

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.
P403+P233 Store in a well-ventilated place. Keep container tightly closed.
P501 Dispose of contents/container in accordance with local regulations.

13. FIRST AID MEASURES

Route of Exposure Action to Be Taken
Inhalation Move to fresh air immediately. If respiratory irritation develops, seek medical attention. Enzyme dust may cause allergic sensitization.
Skin Contact Wash with plenty of soap and water. Remove contaminated clothing. If irritation persists, seek medical attention.
Eye Contact Rinse immediately with plenty of water for at least 15 minutes. Keep eyelids open. Remove contact lenses if present. If irritation persists, seek medical attention.
Ingestion Rinse mouth. Drink plenty of water. If large amounts are swallowed or discomfort is felt, seek medical attention.
Note Enzymes are proteins; allergic reactions are possible in sensitive individuals.

14. FIRE-FIGHTING MEASURES

Parameter Information
Fire Hazard Combustible organic material (enzyme proteins are not highly flammable).
Hazards from Fire Thermal decomposition produces carbon monoxide, carbon dioxide, and nitrogen oxides.
Suitable Extinguishing Media Water spray, CO₂, dry chemical powder, foam.
Unsuitable Extinguishing Media Water jet (may disperse dust)
Special Protective Equipment Self-contained breathing apparatus (SCBA), full protective clothing.
Special Precautions Prevent dust from entering sewers and water courses. Avoid dust cloud formation.

15. ACCIDENTAL RELEASE MEASURES

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.

16. STORAGE AND SHELF LIFE

Parameter All Types
Storage Conditions Store in a cool, dry, well-ventilated place; PROTECT FROM LIGHT, HEAT, and MOISTURE
Container Requirements Keep tightly closed in moisture-proof containers (PE-lined bags, HDPE containers, aluminum-lined bags)
Materials to Protect From Light, heat, moisture, strong oxidizing agents, strong acids, strong bases
Recommended Storage Temperature 4 – 25°C (lower temperature recommended for fungal and plant-based; 4-10°C ideal)
Shelf Life 12 – 24 months (in unopened original packaging under proper storage conditions)
Stability Note Enzyme activity decreases over time; higher temperatures and moisture accelerate activity loss
Special Storage Requirements Store in a dark, cool, dry place; Avoid exposure to air and moisture; Opened containers must be immediately resealed

17. PACKAGING OPTIONS

Packaging Type Quantity Material
Aluminum Foil Bag 100 g, 500 g, 1 kg Aluminum foil laminated bag (moisture and light resistant)
Plastic Bag 1 kg, 5 kg, 10 kg PE bag (inner-lined, sealed, moisture-proof)
PE-Lined Paper Bag 10 kg, 20 kg, 25 kg Moisture-proof kraft paper bag
Plastic Drum 10 kg, 20 kg, 25 kg HDPE drum (moisture-proof)
FIBC / Big Bag 100 kg, 500 kg, 1000 kg PE-lined, moisture-proof PP woven bag
Liquid Container 1 L, 5 L, 10 L, 25 L HDPE / Food-grade plastic

18. TRANSPORT INFORMATION

Parameter Information
UN Number Not regulated (not dangerous)
Hazard Class Not applicable
Packing Group Not applicable
ADR/RID Not regulated
IMDG Code Not regulated (not dangerous)
IATA (Air) Not regulated (not dangerous)
Marine Pollutant No
Transport Temperature Ambient temperature; Protect from moisture and heat.

19. REGULATORY STATUS

Region / Authority Status
European Union Enzyme product approved for use in food and feed; EFSA approval required for specific applications
USA (FDA) GRAS for food use; 21 CFR 173.310 (α-Amylase from Bacillus subtilis)
Turkey Approved as a food processing aid; KKDIK compliance required
JECFA Approved for use as an enzyme preparation; Specifications established for food grade enzymes
IARC Not classified (not carcinogenic)
NTP Not listed
OSHA Regulated as dust (may cause respiratory sensitization)
California (Prop 65) Not listed
Halal Yes (with certification, from halal sources and production)
Kosher Yes (with certification)
Vegan Yes (microbial fermentation, no animal content)
WGK Germany 3 (severe hazard to water)

20. SUMMARY TABLE

Property Fungal Bacterial High Temp. Plant-Based
CAS No 9001-19-8 9000-90-2 9000-90-2 9000-90-2
EC No 3.2.1.1 3.2.1.1 3.2.1.1 3.2.1.1
Source Aspergillus oryzae B. amyloliquefaciens B. licheniformis Barley, wheat, rice
Opt. pH 4.5 – 6.0 6.0 – 7.0 6.0 – 7.0 5.5 – 6.5
Opt. Temp. 50 – 60°C 60 – 75°C 70 – 90°C 30 – 40°C
Inactivation >65-70°C >85-90°C >100-105°C >55-60°C
Mol. Weight ~50-60 kDa ~50-70 kDa ~50-70 kDa ~45-55 kDa
Calcium Moderate High High Low
Mode Endo Endo Endo Endo/Exo
Thermostability Low-Moderate High Very High Low
Main Use Baking Starch industry HFCS, bioethanol Malt, beer

21. CRITICAL NOTICES AND BEST PRACTICES

CRITICAL NOTICES:

  1. CAS Number Verification:

    • Fungal Alpha-Amylase: ✅ 9001-19-8

    • Bacterial Alpha-Amylase: ✅ 9000-90-2

    • High Temperature: ✅ 9000-90-2

    • Plant-Based: ✅ 9000-90-2

    • 9000-85-5 → Old/alternative designation; not valid in current official classifications.

  2. Potential Allergen: Enzyme dust may cause respiratory sensitization; use mask.

  3. Heat Sensitivity: Fungal and plant-based types are more heat-sensitive; pay attention to processing temperatures.

  4. Calcium Requirement: Calcium is important for bacterial and High Temperature types; water quality may affect activity.

  5. Activity Standardization: Activity varies by product grade; always check activity and adjust dosage accordingly.

  6. Optimum Conditions: pH and temperature should be optimized for each application; performance depends on substrate and process conditions.

  7. Inactivation: Fungal types are inactivated during baking (>70°C); bacterial and High Temp. types are inactivated at higher temperatures (>90-105°C).

  8. Vegan and Halal: All types are derived from microbial fermentation and are vegan/halal certified.

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. Enzyme dust may cause allergic sensitization; appropriate precautions should be taken when handling enzyme products.

CAS Number Verification:

  • 9001-19-8 (Fungal Alpha-Amylase - CORRECT)

  • 9000-90-2 (Bacterial, High Temp., Plant-Based - CORRECT)

  • 9000-85-5 (Old/alternative – NOT VALID)

Always verify the correct CAS number and specifications on the supplier's SDS.

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