Glucoamylase, Amyloglucosidase, Glucose Amylase, 9032-08-0

Glucoamylase, Amyloglucosidase, Glucose Amylase, 9032-08-0

GLUCOAMYLASE (AMYLOGLUCOSIDASE)

Glucoamylase / Amyloglucosidase / Glucan 1,4-α-Glucosidase / γ-Amylase / Glucose Amylase

• CAS Number: 9032-08-0

• EC Number: 232-877-2

• Enzyme Class: EC 3.2.1.3

SECTION 1: PRODUCT DEFINITION AND CHEMICAL IDENTITY

Description:
Glucoamylase is an exo-enzyme that hydrolyzes α-1,4 and α-1,6 glycosidic bonds in starch, amylopectin, amylose, glycogen, and malto-oligosaccharides, sequentially releasing β-D-glucose units. Also known as amyloglucosidase or γ-amylase, this enzyme is one of the most important enzymes in industrial starch processing due to its ability to convert starch almost completely to glucose. Unlike alpha-amylase and beta-amylase, glucoamylase can cleave both α-1,4 and α-1,6 bonds, therefore it can completely convert starch to glucose without leaving limit dextrins. It is widely used in the food industry for high-glucose syrup, high-fructose syrup, beer, alcohol, and confectionery production.

SECTION 2: SYNONYMS AND OTHER NAMES

SECTION 3: ENZYMATIC STRUCTURE

3.1. Molecular Structure
Glucoamylase is a glycoprotein consisting of two main domains: the catalytic domain and the starch-binding domain. The catalytic domain has a (α/α)₆-barrel structure, also known as a six-fold alpha-helix/beta-sheet structure. The starch-binding domain (SBD) allows the enzyme to bind effectively to starch granules and carry out efficient hydrolysis. The active site contains aspartate and glutamate residues that play critical roles in the catalytic mechanism.

3.2. Catalytic Mechanism
Glucoamylase binds to the non-reducing end of the starch chain and carries out hydrolysis of glycosidic bonds through acid-base catalysis. The enzyme can cleave both α-1,4 bonds and α-1,6 branching bonds, making it ideal for complete conversion of starch to glucose. Carboxylate groups in the catalytic site attack the glycosidic bond of the substrate, releasing β-D-glucose units.

3.3. Molecular Weight
60,000 - 100,000 Da (varies by source; fungal sources typically 70,000-100,000 Da)

3.4. Basic Structural Properties

SECTION 4: PHYSICAL AND CHEMICAL PROPERTIES

SECTION 5: CHEMICAL REACTION

5.1. Catalyzed Reaction
Starch + n H₂O → n/2 C₆H₁₂O₆ (β-D-Glucose)

5.2. Reaction Mechanism
Glucoamylase removes successive glucose units from the non-reducing end of the starch chain. Unlike alpha-amylase, glucoamylase can hydrolyze both α-1,4 bonds and α-1,6 branching bonds. This property allows glucoamylase to convert starch to glucose with over 95% yield. As the reaction progresses and the chain shortens, the enzyme's affinity for the substrate changes, and hydrolysis of smaller oligosaccharides proceeds more slowly.

5.3. Substrate Specificity

SECTION 6: PHYSICAL AND CHEMICAL PROPERTIES TABLE

SECTION 7: ENZYMATIC ACTIVITY AND REACTION

SECTION 8: REACTION EQUATIONS

8.1. Complete Hydrolysis of Starch (General Reaction)
(C₆H₁₀O₅)n + n H₂O → n C₆H₁₂O₆ (β-D-Glucose)

8.2. Hydrolysis of Amylose
Amylose + H₂O → β-D-Glucose

8.3. Hydrolysis of Amylopectin
Amylopectin + H₂O → β-D-Glucose (including branch points)

8.4. Hydrolysis of Maltose
Maltose (C₁₂H₂₂O₁₁) + H₂O → 2 C₆H₁₂O₆ (Glucose)

8.5. Enzyme Inactivation (Thermal)
Glucoamylase (active) → Glucoamylase (inactive) (≥70°C, 10-15 minutes)

SECTION 9: PRODUCTION METHOD

Glucoamylase is primarily produced through microbial fermentation.

Alternative Production Method (Recombinant): High-yield glucoamylase production is possible using genetically modified Aspergillus or Saccharomyces species. Recombinant glucoamylases provide higher purity, specific activity, and thermal stability.

SECTION 10: APPLICATIONS AND INDUSTRIAL USES

10.1. Starch Processing and Glucose Syrup Production

Example Recipe – High-Glucose Syrup:

• Liquefied starch (30-35% dry solids): 1000 L

• Glucoamylase: 100-500 g (depending on activity)

• pH: 4.2-4.5

• Temperature: 55-60°C

• Reaction time: 24-72 hours

Adjust liquefied starch to pH 4.2-4.5 and heat to 55-60°C. Add glucoamylase and incubate until DE (Dextrose Equivalent) value reaches 95-98 (24-72 hours). Inactivate the enzyme by heat treatment and purify the syrup.

10.2. Beer and Alcohol Production

10.3. Confectionery and Food Industry

10.4. Other Applications

SECTION 11: ALTERNATIVES AND COMPARISON

11.1. Alpha-Amylase (EC 3.2.1.1)

11.2. Beta-Amylase (EC 3.2.1.2)

11.3. Pullulanase (EC 3.2.1.41)

11.4. Glucose Isomerase (EC 5.3.1.18)

SECTION 12: TOXICOLOGY AND SAFETY

12.1. Acute Toxicity

12.2. Regulatory Status

SECTION 13: GHS CLASSIFICATION (Powder Form)

Signal Word: Danger

Hazard Pictograms: GHS07 (Exclamation mark), GHS08 (Health Hazard)

Hazard Statements (H-Codes):

• H334: May cause allergy or asthma symptoms if inhaled

• H319: Causes serious eye irritation

• H335: May cause respiratory irritation

Precautionary Statements (Historical):

SECTION 14: PRECAUTIONARY STATEMENTS (P-CODES)

SECTION 15: FIRST AID MEASURES

SECTION 16: FIREFIGHTING MEASURES

SECTION 17: ACCIDENTAL RELEASE MEASURES

SECTION 18: STORAGE AND SHELF LIFE

SECTION 19: PACKAGING OPTIONS

SECTION 20: TRANSPORT INFORMATION

SECTION 21: ENVIRONMENTAL INFORMATION

SECTION 22: REGULATORY STATUS

SECTION 23: FREQUENTLY ASKED QUESTIONS (FAQ)

Q1: Is glucoamylase safe?
A1: Yes. Glucoamylase is classified as GRAS (Generally Recognized As Safe) by the FDA and approved as a food enzyme by EFSA. However, the powder form may cause allergic reactions when inhaled, so appropriate personal protective equipment should be used.

Q2: What is the main difference between glucoamylase and alpha-amylase?
A2: Alpha-amylase is an endo-enzyme that randomly cleaves the interior of the starch molecule for liquefaction. Glucoamylase is an exo-enzyme that removes successive glucose units from the chain ends. Glucoamylase can cleave both α-1,4 and α-1,6 bonds, therefore it can completely convert starch to glucose.

Q3: Is glucoamylase suitable for vegan and halal products?
A3: Microbial (fungal or bacterial) sourced glucoamylases are suitable for vegan and halal certification. Since the product is produced from microorganisms such as Aspergillus, it contains no animal ingredients. Relevant certificates can be obtained from manufacturers.

Q4: What is the shelf life of glucoamylase?
A4: In unopened original packaging, stored in a cool (4-25°C) and dry place away from direct sunlight, a shelf life of 24 months is recommended. For long-term storage, storing at 4°C minimizes activity loss.

Q5: How should glucoamylase be stored?
A5: Store in a cool (preferably 4-25°C), dry, well-ventilated area in tightly closed containers. Moisture, high temperature (>40°C), and direct sunlight cause activity loss. Contact with heavy metals (copper, zinc, iron) should be avoided.

Q6: What are the optimum working conditions for glucoamylase?
A6: The optimum pH of glucoamylase is between 4.0-5.0, and the optimum temperature varies between 55-65°C depending on the source. Fungal enzymes typically show best activity around 55-60°C. The enzyme is rapidly inactivated above 70°C.

Q7: Which factors inhibit glucoamylase?
A7: Glucoamylase is inhibited by heavy metals (Hg²⁺, Ag⁺, Cu²⁺), gluconic acid, SDS, and β-cyclodextrin. Calcium (Ca²⁺) ions activate enzyme activity in some forms. Glucose (product inhibition) slows enzyme activity at high concentrations.

Q8: Why is glucoamylase important in high-glucose syrup production?
A8: Glucoamylase is the only enzyme that can convert alpha-amylase-liquefied starch to glucose with over 95% yield. With its ability to cleave both α-1,4 and α-1,6 bonds, it breaks down limit dextrins left by other amylases, providing maximum glucose yield.

Q9: How is high-glucose syrup produced with glucoamylase?
A9: For high-glucose syrup production, starch is first liquefied with alpha-amylase (80-105°C). Then the pH is adjusted to 4.2-4.5, cooled to 55-60°C, and glucoamylase is added. After 24-72 hours of incubation, the DE (Dextrose Equivalent) value reaches 95-98. The enzyme is inactivated by heat treatment and the syrup is purified.

Q10: What are the effects of inhaling glucoamylase powder?
A10: Glucoamylase powder, when inhaled, can cause allergic reactions, asthma-like symptoms, and respiratory tract irritation. Occupational asthma known as "baker's asthma" can develop especially in baking and food processing industries upon repeated exposure to enzyme dust. Therefore, a dust mask (P2/P3) must always be used when handling the powder form, and local exhaust ventilation should be provided.

SECTION 24: QUICK REFERENCE TABLE

SECTION 25: CRITICAL WARNINGS AND BEST PRACTICES

CRITICAL WARNINGS:

1. Dust Control and Respiratory Sensitization: Glucoamylase powder can cause occupational asthma (baker's asthma) upon repeated exposure. Use local exhaust ventilation to prevent dust dispersion and always wear a dust mask (P2/P3). Liquid formulations are safer than powder forms.

2. Activity Loss - Temperature: Glucoamylase is rapidly and irreversibly inactivated above 70°C. Strictly maintain application temperatures within the optimum range (55-60°C). Temperature control is critical in saccharification processes.

3. Activity Loss - pH: Glucoamylase loses stability below pH 3.5 or above pH 6.0. Maintain application pH within the optimum range (4.0-5.0). Excessively acidic or basic conditions cause permanent enzyme inactivation.

4. Activity Loss - Heavy Metals: Mercury (Hg²⁺), silver (Ag⁺), and copper (Cu²⁺) ions inhibit glucoamylase. Avoid these metals in equipment that comes into contact with the enzyme. Stainless steel, HDPE, or glass materials are preferred.

5. Food Grade: Only FDA/EFSA-approved food-grade glucoamylase meeting specifications should be used in food applications. Industrial grade enzymes may contain impurities and are not suitable for human consumption.

6. Product Inhibition: High concentrations of glucose (product) inhibit enzyme activity. Process conditions should be optimized to ensure glucose concentration does not exceed certain levels during the reaction.

7. α-1,6 Bond Hydrolysis: Glucoamylase hydrolyzes α-1,6 bonds more slowly than α-1,4 bonds. Sufficient reaction time should be provided for complete conversion. If needed, combination with debranching enzymes such as pullulanase can be considered.

BEST PRACTICE RECOMMENDATIONS:

• Storage: Store in a cool (4-25°C), dry, well-ventilated area in tightly closed containers. For long-term storage, +4°C (refrigeration) is recommended. Use within a short period after opening.

• Handling: Use protective gloves, dust mask (P2/P3), and protective goggles. To prevent dust dispersion, preferably use liquid formulations.

• Dosage: Calculate the correct dosage according to enzyme activity. Check enzyme activity for each batch. Overdosing may adversely affect product quality and increase costs.

• Enzyme Inactivation: Inactivate the enzyme by heat treatment (≥70°C, 10-15 minutes, pH < 3.5) at the end of the process. This prevents unwanted enzymatic activity from affecting product quality.

• Quality Control: Perform activity testing for each batch. Maintain production and handling hygiene to prevent microbiological contamination. Perform regular DE value and sugar profile analysis.

• Waste Management: Dispose of enzyme and contaminated materials in accordance with local regulations. Do not discharge into sewers.

LEGAL DISCLAIMER:

This Technical Data Sheet (TDS) is for informational purposes only and is prepared based on available technical data. The user is 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, refer to the official Safety Data Sheet (SDS/MSDS) provided by the manufacturer/supplier. Glucoamylase powder may cause sensitization when inhaled; appropriate protective equipment should be used. This document does not replace professional or medical advice.