Lipase, Triacyl Glycerol Lipase, Fat-Splitting Enzyme, Glycerol Ester Hydrolase, 9001-62-1

Lipase, Triacyl Glycerol Lipase, Fat-Splitting Enzyme, Glycerol Ester Hydrolase, 9001-62-1

LIPASE (LIPASE ENZYME)

Lipase / Triacylglycerol Lipase / Fat-Splitting Enzyme / Glycerol Ester Hydrolase

• CAS Number: 9001-62-1

• EC Number: 232-619-9

• Enzyme Class: EC 3.1.1.3

SECTION 1: PRODUCT DEFINITION AND CHEMICAL IDENTITY

Description:
Lipase is an enzyme that hydrolyzes triglycerides (fats) into free fatty acids and glycerol. Lipases are active at the oil-water interface and exhibit a unique property known as "interfacial activation." This enzyme plays a critical role in fat metabolism in biological systems. In the food industry, it is widely used for flavor development in cheese production, fat hydrolysis, and improvement of taste and texture. It is also used in the pharmaceutical industry for digestive enzyme formulations, in the detergent industry as a fat stain remover, and in biodiesel production.

SECTION 2: SYNONYMS AND OTHER NAMES

SECTION 3: ENZYMATIC STRUCTURE

3.1. Molecular Structure
Lipase is a globular protein consisting of a single polypeptide chain. The enzyme's active site has a characteristic three-dimensional structure known as the α/β hydrolase fold. The active site contains a catalytic triad consisting of serine-histidine-aspartate/glutamate. Some lipases have a "lid" domain that covers the active site and opens during interfacial activation. This lid domain enables the enzyme's activation at the oil-water interface.

3.2. Catalytic Mechanism
Lipase catalyzes the hydrolysis of triglycerides at the oil-water interface. The catalytic mechanism proceeds similarly to serine proteases:

1. Interfacial Activation: Lipase binds to the oil-water interface and the lid domain opens, making the active site accessible to the substrate.

2. Catalytic Attack: The serine residue in the active site performs a nucleophilic attack on the ester bond of the triglyceride molecule.

3. Intermediate Formation: An acyl-enzyme intermediate is formed.

4. Deacylation: A water molecule attacks the acyl-enzyme intermediate, releasing free fatty acid and glycerol.

3.3. Molecular Weight
30,000 - 60,000 Da (varies by source; fungal sources ~30,000-40,000 Da; bacterial sources ~30,000-60,000 Da; pancreatic lipase ~48,000 Da)

3.4. Basic Structural Properties

SECTION 4: PHYSICAL AND CHEMICAL PROPERTIES

SECTION 5: SUBSTRATE SPECIFICITY

SECTION 6: ENZYMATIC ACTIVITY AND REACTION

SECTION 7: REACTION MECHANISM

Lipase catalyzes the hydrolysis of triglycerides through a mechanism involving the following steps:

7.1. Interfacial Activation
Lipase binds to the oil-water interface. During this binding, the enzyme's lid domain opens and the active site becomes accessible to the substrate. This phenomenon is known as "interfacial activation."

7.2. Substrate Binding
The triglyceride molecule binds to the active site. Fatty acid chains are positioned into the hydrophobic pockets of the enzyme.

7.3. Catalytic Attack
The serine residue (Ser) in the active site performs a nucleophilic attack on the ester bond of the triglyceride. The histidine (His) residue deprotonates serine, making it a stronger nucleophile. The aspartate/glutamate (Asp/Glu) residue stabilizes the protonation state of histidine.

7.4. Acyl-Enzyme Intermediate
The nucleophilic attack results in the formation of a tetrahedral intermediate. The intermediate collapses, forming an acyl-enzyme intermediate and diglyceride.

7.5. Deacylation
A water molecule attacks the acyl-enzyme intermediate, releasing free fatty acid and regenerating the active enzyme.

7.6. Positional Specificity

• 1,3-Specific Lipases: Hydrolyze ester bonds at positions 1 and 3 of triglycerides. Products: 2-monoglyceride and free fatty acids.

• Non-Specific Lipases: Hydrolyze all ester bonds at positions 1, 2, and 3 of triglycerides. Products: glycerol and free fatty acids.

SECTION 8: REACTION EQUATIONS

8.1. Triglyceride Hydrolysis (General Reaction)
Triglyceride + 3 H₂O → Glycerol + 3 Free Fatty Acids

8.2. 1,3-Specific Lipase Reaction
Triglyceride + 2 H₂O → 2-Monoglyceride + 2 Free Fatty Acids

8.3. Non-Specific Lipase Reaction
Triglyceride + 3 H₂O → Glycerol + 3 Free Fatty Acids

8.4. Partial Hydrolysis (Diglyceride Formation)
Triglyceride + H₂O → Diglyceride + Free Fatty Acid

8.5. Enzyme Inactivation (Thermal)
Lipase (active) → Lipase (inactive) (≥60°C, 10-15 minutes)

8.6. Enzyme Inhibition (Orlistat)
Lipase + Orlistat → Lipase-Orlistat Complex (inactive)

SECTION 9: LIPASE TYPES

SECTION 10: APPLICATIONS AND INDUSTRIAL USES

10.1. Cheese and Dairy Industry

Example Recipe – Cheese Flavor Development:

• Milk: 1000 L

• Lipase: 1-10 g (depending on activity)

• Application temperature: 35-40°C

• Incubation time: 1-4 hours

Heat milk to 35-40°C. Dissolve lipase in a small amount of milk and add to the mixture. Incubate for the specified time. Continue with cheese making.

10.2. Oil and Fat Industry

10.3. Baking and Cereal Industry

10.4. Detergent Industry

10.5. Pharmaceutical and Nutraceutical Industry

10.6. Biodiesel Production

10.7. Other Industrial Applications

SECTION 11: ALTERNATIVES AND COMPARISON

11.1. Chemical Hydrolysis (Acid or Base)

11.2. Esterase (EC 3.1.1.1)

11.3. Chymotrypsin (EC 3.4.21.1)

11.4. Phospholipase (EC 3.1.1.4)

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

Safety 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 lipase safe for food applications?
A1: Yes. Lipase is classified as GRAS (Generally Recognized As Safe) by the FDA and approved as a food enzyme by EFSA. It is widely used in cheese production, baking, and fat modification. However, the powder form may cause allergic reactions when inhaled, so appropriate personal protective equipment should be used.

Q2: What is the role of lipase in fat hydrolysis?
A2: Lipase hydrolyzes triglycerides (fats) into free fatty acids and glycerol. This hydrolysis releases short-chain fatty acids (butyric, capric, caprylic acid), which are responsible for characteristic flavor development, especially in cheese production. Additionally, it modifies the functional properties of fats and supports flavor development.

Q3: What is the difference between lipase and esterase?
A3: Lipase is active at the oil-water interface and hydrolyzes triglycerides (fats). It exhibits interfacial activation. Esterase hydrolyzes water-soluble esters, does not hydrolyze triglycerides (or very little), and does not require interfacial activation. Lipases are suitable for large, fatty substrates, while esterases are suitable for small, water-soluble esters.

Q4: Is lipase suitable for vegan and halal products?
A4: Microbial (fungal or bacterial) sourced lipases are suitable for vegan and halal certification. However, animal-derived pancreatic lipases are not vegan and require special approval for halal certification. Bacterial and fungal lipases are preferred for vegan and halal products as they contain no animal ingredients. Relevant certificates can be obtained from manufacturers.

Q5: What is the shelf life of lipase?
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. Formulations containing calcium ions provide better stability.

Q6: How should lipase 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 (copper, mercury) and metal chelators (EDTA) should be avoided.

Q7: What are the optimum working conditions for lipase?
A7: The optimum pH of lipase is between 6.0-8.0, and the optimum temperature varies between 30-45°C depending on the source. The enzyme is stable up to 50°C and rapidly inactivated above 60°C. Some lipases require calcium (Ca²⁺) ions. It is active at the oil-water interface, so emulsion formation is important for activity.

Q8: Why is lipase used in cheese production?
A8: In cheese production, lipase hydrolyzes milk fat to produce short-chain fatty acids (butyric, capric, caprylic acid). These fatty acids give cheese its characteristic sharp and pungent flavor. Lipase use is especially common in Italian cheeses such as Parmesan, Romano, and Provolone. Lipase is also used in enzyme-modified cheese (EMC) production to obtain concentrated cheese flavor.

Q9: How is lipase used in biodiesel production?
A9: In biodiesel production, lipase catalyzes the transesterification reaction that converts triglycerides (fats) to fatty acid methyl esters (biodiesel). Enzymatic biodiesel production offers milder conditions (low temperature, normal pressure), fewer byproducts, and higher purity glycerol byproduct compared to conventional chemical methods. However, enzyme cost and reaction time are disadvantages compared to chemical methods.

Q10: What are the effects of inhaling lipase powder?
A10: Lipase 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. Liquid formulations are safer alternatives.

SECTION 24: QUICK REFERENCE TABLE

SECTION 25: CRITICAL WARNINGS AND BEST PRACTICES

CRITICAL WARNINGS:

1. Dust Control and Respiratory Sensitization: Lipase 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. Interfacial Activation: Lipase is active at the oil-water interface. The substrate (fat) must be in emulsion form for optimal enzyme activity. Homogenization or emulsion formation significantly increases activity.

3. Activity Loss - Temperature: Lipase is rapidly and irreversibly inactivated above 60°C. Strictly maintain application temperatures within the optimum range (30-45°C). Temperature control is critical in cheese production and fat modification.

4. Activity Loss - pH: Lipase loses stability below pH 5.0 or above pH 9.0. Maintain application pH within the optimum range (6.0-8.0). Excessively acidic or basic conditions cause permanent enzyme inactivation.

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

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

7. Source Origin: Microbial (fungal or bacterial) sources are suitable for vegan and halal products. Animal-derived lipases (pancreatic) are not suitable for vegan applications and require additional certification for halal. Always verify source with the manufacturer.

8. Positional Specificity: Different lipases show different positional specificity. 1,3-specific lipases hydrolyze only positions 1 and 3, while non-specific lipases hydrolyze all positions. Select the correct lipase type 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. Calcium-containing formulations have better stability.

• 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.

• Emulsion Formation: Substrate (fat) must be in emulsion form for lipase activity. Formulate emulsions using homogenization or high-shear mixing. Emulsifiers (e.g., lecithin) may be used.

• 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 0.001-0.01% of milk for cheese applications and 0.1-1% of formulation for detergent applications.

• Enzyme Inactivation: Inactivate the enzyme by heat treatment (≥60°C, 10-15 minutes) at the end of the process to prevent over-hydrolysis. For cheese products, pasteurization or other heat treatments will inactivate residual enzyme activity.

• Calcium Addition: Consider adding calcium ions (CaCl₂) to the application solution to improve enzyme stability and activity, particularly in calcium-free systems.

• Quality Control: Perform activity testing for each batch. Maintain production and handling hygiene to prevent microbiological contamination. Monitor fatty acid profile and degree of hydrolysis 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. Lipase powder may cause sensitization when inhaled; appropriate protective equipment should be used. This document does not replace professional or medical advice.