Invertase, Sucrase, Beta-Fructofuranosidase, Saccharase, Invert Sugar Enzyme, 9001-57-4

Invertase, Sucrase, Beta-Fructofuranosidase, Saccharase, Invert Sugar Enzyme, 9001-57-4

INVERTASE (SUCRASE, BETA-FRUCTOFURANOSIDASE)

Invertase / Sucrase / Beta-Fructofuranosidase / Saccharase / Invert Sugar Enzyme

• CAS Number: 9001-57-4

• EC Number: 232-608-5

• Enzyme Class: EC 3.2.1.26

SECTION 1: PRODUCT DEFINITION AND CHEMICAL IDENTITY

Description:
Invertase is an enzyme that hydrolyzes sucrose into glucose and fructose. The resulting mixture of glucose and fructose is called "invert sugar." Invertase is also known as beta-fructofuranosidase or sucrase. The enzyme hydrolyzes the α,β-1,2-glycosidic bond in the sucrose molecule, releasing glucose and fructose. In the food industry, it is widely used especially in confectionery and dessert production for invert sugar production, preventing sugar crystallization, increasing moisture retention capacity, and improving taste and texture. It is also used in honey production, brewing, and invert sugar syrup production.

SECTION 2: SYNONYMS AND OTHER NAMES

SECTION 3: ENZYMATIC STRUCTURE

3.1. Molecular Structure
Invertase is a globular glycoprotein consisting of a polypeptide chain. Invertase from yeast typically contains 40-60% carbohydrate (mannan). The enzyme has two main active sites that catalyze the hydrolysis of glycosidic bonds. The carbohydrate portion of the molecule increases the enzyme's stability at high temperatures and under challenging conditions. Invertase exists in extracellular and intracellular forms. The extracellular form has higher molecular weight and is more highly glycosylated.

3.2. Catalytic Mechanism
Invertase hydrolyzes sucrose through an acid-base catalysis mechanism:

1. Substrate Binding: The sucrose molecule binds to the active site of the enzyme. Carboxylate groups in the active site approach the glycosidic bond of the substrate.

2. Catalytic Attack: A glutamic acid residue in the active site donates a proton to the glycosidic bond. This causes the fructose moiety to leave and forms a carbon cation intermediate.

3. Nucleophilic Attack: A water molecule attacks the carbon cation intermediate, releasing glucose and fructose.

4. Product Release: The glucose and fructose products are released from the active site.

3.3. Molecular Weight
50,000 - 60,000 Da (from yeast, monomer); 100,000 - 500,000 Da (aggregate forms)

3.4. Basic Structural Properties

SECTION 4: PHYSICAL AND CHEMICAL PROPERTIES

SECTION 5: ENZYMATIC ACTIVITY AND REACTION

SECTION 6: REACTION MECHANISM

Invertase catalyzes the hydrolysis of sucrose through a mechanism involving the following steps:

6.1. Substrate Binding
The sucrose molecule (α-D-glucose-β-D-fructose) binds to the active site of the enzyme. Binding is determined by the enzyme's affinity for the substrate.

6.2. Proton Transfer
A glutamic acid residue in the active site donates a proton to the oxygen atom of the glycosidic bond. This weakens the bond and causes the fructose moiety to leave.

6.3. Carbon Cation Formation
Cleavage of the glycosidic bond creates a carbon cation intermediate on the glucose moiety. This intermediate is stabilized in the enzyme's active site.

6.4. Nucleophilic Attack
A water molecule (nucleophile) attacks the carbon cation intermediate, releasing glucose and fructose. In this step, α-D-glucose and β-D-fructose are formed.

6.5. Product Release
The glucose and fructose products are released from the active site, and the enzyme becomes ready for another catalytic cycle.

SECTION 7: REACTION EQUATIONS

7.1. Sucrose Hydrolysis (Main Reaction)
C₁₂H₂₂O₁₁ (Sucrose) + H₂O → C₆H₁₂O₆ (Glucose) + C₆H₁₂O₆ (Fructose)

7.2. Invert Sugar Formation
Sucrose + H₂O (Invertase) → Invert Sugar (Glucose + Fructose)

7.3. Raffinose Hydrolysis (Partial)
Raffinose + H₂O → Melibiose + Fructose

7.4. Enzyme Inactivation (Thermal)
Invertase (active) → Invertase (inactive) (≥65°C, 10-15 minutes)

7.5. Product Inhibition
Invertase + Glucose → Invertase-Glucose Complex (inactive)

SECTION 8: INVERTASE TYPES

SECTION 9: APPLICATIONS AND INDUSTRIAL USES

9.1. Confectionery and Dessert Production

Example Recipe – Invert Sugar Syrup Production:

• Sucrose: 100 kg

• Water: 50 L

• Invertase: 10-50 g (depending on activity)

• Application temperature: 50-55°C

• pH: 4.5-5.0

• Incubation time: 2-4 hours

Dissolve sucrose in water and heat to 50-55°C. Dissolve invertase in a small amount of water and add to the mixture. Maintain pH at 4.5-5.0. Incubate for the specified time. Invert sugar syrup is obtained. Inactivate the enzyme by heat treatment.

9.2. Confectionery Products (Candies, Caramels, Fondant)

Example Recipe – Fondant Crystallization Prevention:

• Sugar syrup (sucrose): 100 kg

• Invertase: 1-5 g (depending on activity)

• Application temperature: 55°C

• Incubation time: 60-120 minutes

Heat sugar syrup to 55°C. Dissolve invertase in a small amount of water and add to the mixture. Incubate for the specified time. Invertase converts a portion of sucrose to invert sugar, preventing crystallization.

9.3. Honey and Bee Products

9.4. Brewing and Fermentation Industry

9.5. Baking and Cereal Industry

9.6. Pharmaceutical and Nutraceutical Industry

9.7. Other Applications

SECTION 10: ALTERNATIVES AND COMPARISON

10.1. Acid Hydrolysis (Chemical)

10.2. Glucose Isomerase (EC 5.3.1.18)

10.3. Amylase (EC 3.2.1.1)

10.4. Lactase (EC 3.2.1.108)

SECTION 11: TOXICOLOGY AND SAFETY

11.1. Acute Toxicity

11.2. Regulatory Status

SECTION 12: 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

Safety Statements (Historical):

SECTION 13: PRECAUTIONARY STATEMENTS (P-CODES)

SECTION 14: FIRST AID MEASURES

SECTION 15: FIREFIGHTING MEASURES

SECTION 16: ACCIDENTAL RELEASE MEASURES

SECTION 17: STORAGE AND SHELF LIFE

SECTION 18: PACKAGING OPTIONS

SECTION 19: TRANSPORT INFORMATION

SECTION 20: ENVIRONMENTAL INFORMATION

SECTION 21: REGULATORY STATUS

SECTION 22: FREQUENTLY ASKED QUESTIONS (FAQ)

Q1: Is invertase safe for food applications?
A1: Yes. Invertase is classified as GRAS (Generally Recognized As Safe) by the FDA and approved as a food enzyme by EFSA. It is widely used in confectionery, desserts, and bakery products. However, the powder form may cause allergic reactions when inhaled, so appropriate personal protective equipment should be used.

Q2: What is the effect of invertase on sucrose?
A2: Invertase hydrolyzes sucrose into glucose and fructose. The resulting mixture of glucose and fructose is called "invert sugar." Invert sugar is sweeter than sucrose, more hygroscopic, and does not crystallize. Therefore, it is preferred in confectionery and dessert production.

Q3: What is the difference between invertase and acid hydrolysis?
A3: Invertase provides enzymatic hydrolysis; it is specific, works under mild conditions, and produces no byproducts. Acid hydrolysis is a chemical method; it requires high temperature, can produce byproducts (HMF), and may cause corrosion. Enzymatic hydrolysis is preferred for high-quality invert sugar production.

Q4: Is invertase suitable for vegan and halal products?
A4: Microbial (yeast or fungal) sourced invertases are suitable for vegan and halal certification. Since the product is produced from microorganisms such as Saccharomyces cerevisiae or Aspergillus, it contains no animal ingredients. Relevant certificates can be obtained from manufacturers.

Q5: What is the shelf life of invertase?
A5: In unopened original packaging, stored in a cool (4-25°C) and dry place away from direct sunlight, a shelf life of 12-24 months is recommended. For long-term storage, storing at 4°C minimizes activity loss.

Q6: How should invertase be stored?
A6: 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 (silver, copper, mercury) should be avoided.

Q7: What are the optimum working conditions for invertase?
A7: The optimum pH of invertase is between 4.5-5.5, and the optimum temperature varies between 50-60°C depending on the source. Yeast invertases typically show best activity around 55-60°C. The enzyme is stable up to 60°C and rapidly inactivated above 65°C. Substrate inhibition is observed at high substrate concentrations.

Q8: Why is invertase used in confectionery production?
A8: In confectionery production, invertase converts a portion of sucrose to invert sugar, preventing sugar crystallization. This keeps the product smooth, creamy, and homogeneous. Additionally, the hygroscopic property of invert sugar keeps the product moist and extends shelf life. It is widely used in fondant, caramel, marshmallow, and candy production.

Q9: How is invertase used in invert sugar syrup production?
A9: For invert sugar syrup production, a sucrose solution is prepared, adjusted to pH 4.5-5.0, and heated to 50-55°C. Invertase is added and incubated for 2-4 hours. Sucrose is hydrolyzed to glucose and fructose. The enzyme is inactivated by heat treatment, and invert sugar syrup is obtained. This syrup is used in confectionery, beverage, and pharmaceutical industries.

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

SECTION 23: QUICK REFERENCE TABLE

SECTION 24: CRITICAL WARNINGS AND BEST PRACTICES

CRITICAL WARNINGS:

1. Dust Control and Respiratory Sensitization: Invertase powder can cause occupational 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. Substrate Inhibition: Invertase shows substrate inhibition at high sucrose concentrations (>1M). Conduct experiments to determine optimum substrate concentration. Excessively high sucrose concentrations reduce enzyme activity.

3. Activity Loss - Temperature: Invertase is rapidly and irreversibly inactivated above 65°C. Strictly maintain application temperatures within the optimum range (50-60°C). Temperature control is critical in confectionery and invert sugar production.

4. Activity Loss - pH: Invertase loses stability below pH 3.5 or above pH 6.5. Maintain application pH within the optimum range (4.5-5.5). Excessively acidic or basic conditions cause permanent enzyme inactivation.

5. Product Inhibition: Glucose inhibits invertase activity (product inhibition). Reaction rate decreases at high glucose concentrations. Activity is higher in continuous systems or processes where product is removed.

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

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

8. Glycosylation and Stability: Yeast invertase is highly glycosylated and therefore more stable. Fungal invertases have lower glycosylation levels and work better at lower temperatures. Select the correct source for your application.

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. Prepare working solutions fresh for each application.

• Substrate Preparation: Mix the sucrose solution thoroughly and ensure homogeneity. Maintain substrate concentration at optimum level. Pre-heating facilitates sucrose hydrolysis by the enzyme.

• Dosage: Calculate the correct dosage according to enzyme activity. Check enzyme activity for each batch. Start with low dosage and adjust based on results. Typical dosage ranges are 0.01-0.1% for invert sugar production and 0.005-0.02% for confectionery applications.

• Enzyme Inactivation: Inactivate the enzyme by heat treatment (≥65°C, 10-15 minutes) at the end of the process to prevent over-hydrolysis. For invert sugar syrup, pasteurization will inactivate residual enzyme activity.

• Quality Control: Perform activity testing for each batch. Maintain production and handling hygiene to prevent microbiological contamination. Monitor invert sugar content, sweetness level, and crystallization status during processing for consistent product quality.

• Waste Management: Dispose of enzyme and contaminated materials in accordance with local regulations. Do not discharge into sewers. If released into wastewater, inactivate the enzyme by heat treatment before disposal.

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. Invertase powder may cause sensitization when inhaled; appropriate protective equipment should be used. This document does not replace professional or medical advice.