Beta Amylase, Glucanohydrolase, Glucanase, Glucan Maltohydrolase, 9000-91-3

Beta Amylase, Glucanohydrolase, Glucanase, Glucan Maltohydrolase, 9000-91-3

BETA-AMYLASE (β-Amylase)

Beta-Amylase / β-Amylase / Glucanohydrolase / 1,4-α-D-Glucan Maltohydrolase

• CAS Number: 9000-91-3

• EC Number: 232-566-1

• Enzyme Class: EC 3.2.1.2

SECTION 1: PRODUCT DEFINITION AND CHEMICAL IDENTITY

Description:
Beta-Amylase is an exo-enzyme that hydrolyzes α-1,4-glycosidic bonds in polysaccharides such as starch and glycogen, sequentially removing maltose units from the non-reducing ends of the chain. It occurs naturally in plants, bacteria, and fungi. In the food industry, it plays a critical role in beer production (mashing), baking (dough fermentation), and high-purity maltose syrup production. Unlike alpha-amylase, it does not randomly cleave the interior of the starch molecule; instead, it systematically removes maltose units from the chain ends.

SECTION 2: SYNONYMS AND OTHER NAMES

SECTION 3: ENZYMATIC STRUCTURE

3.1. Molecular Structure
Beta-Amylase is a globular protein consisting of a polypeptide chain. Its secondary structure contains alpha-helix and beta-sheet motifs. The active site contains specific amino acid residues (particularly aspartate and glutamate) that bind to the terminal end of the starch molecule and carry out catalytic activity. Disulfide bonds in the enzyme's structure have a significant effect on thermal stability.

3.2. Catalytic Mechanism
The enzyme binds to the non-reducing end of the starch chain and hydrolyzes the glycosidic bond through acid-base catalysis. Carboxylate groups in the catalytic site attack the glycosidic bond of the substrate, resulting in the cleavage of maltose units.

3.3. Molecular Weight
30,000 - 60,000 Da (varies by source; plant ~50,000-60,000 Da, bacterial ~30,000-35,000 Da)

3.4. Basic Structural Properties

SECTION 4: PHYSICAL AND CHEMICAL PROPERTIES

SECTION 5: SPECIFICATIONS (TYPICAL FOOD GRADE)

SECTION 6: ENZYMATIC ACTIVITY AND REACTION

SECTION 7: REACTION MECHANISM

Beta-Amylase catalyzes the hydrolysis of successive maltose units from the non-reducing end of the starch chain. This process can occur through two different mechanisms:

7.1. Multi-Chain Attack Mechanism:
The enzyme binds to one substrate molecule, cleaves one maltose unit, and then moves to a new substrate molecule. This mechanism allows the enzyme to move rapidly between different starch chains.

7.2. Single-Chain Attack Mechanism:
The enzyme remains on the same substrate chain and sequentially cleaves multiple maltose units as it progresses. This mechanism provides higher maltose yield but is slower.

7.3. Limit Dextrin Formation:
Beta-Amylase cannot cleave the α-1,6 bonds at the branching points of amylopectin. Therefore, at the end of the reaction, a high molecular weight residue called "limit dextrin" remains. This limit dextrin consists of short side chains located near the branching points.

SECTION 8: REACTION EQUATIONS

8.1. Hydrolysis of Starch (General Reaction)
(C₆H₁₀O₅)n + n H₂O → n/2 C₁₂H₂₂O₁₁ (Maltose)

8.2. Hydrolysis of Amylose
Amylose + H₂O → Maltose + Limit Dextrin

8.3. Hydrolysis of Amylopectin
Amylopectin + H₂O → Maltose + Limit Dextrin (branch points remain)

8.4. Enzyme Inactivation (Thermal)
Beta-Amylase (active) → Beta-Amylase (inactive) (≥70°C)

SECTION 9: PRODUCTION METHOD

Beta-Amylase is primarily produced through microbial fermentation or plant extraction.

Alternative Production Method (Recombinant): High-yield beta-amylase production is possible using genetically modified microorganisms. This method provides higher purity and specific activity.

SECTION 10: APPLICATIONS AND INDUSTRIAL USES

10.1. Beer Production (Malting and Brewing)

Example Recipe – Beer Mashing:

• Malt: 100 kg

• Water: 300 L

• Beta-Amylase: 5-10 g (depending on activity)

• Mashing temperature: 62-65°C, 60-90 minutes

Grind the malt, mix with water, and bring to mashing temperature. Add beta-amylase and carry out mashing for the specified time. Filter the mash and collect the wort.

10.2. Baking (Bread, Biscuits, Crackers)

Example Recipe – Sandwich Bread:

• Flour: 100 kg

• Water: 60 L

• Yeast: 2 kg

• Salt: 1.5 kg

• Beta-Amylase: 5-10 g (depending on activity)

Mix all dry ingredients, add water and yeast. Knead the dough, allow to rest, shape, and bake.

10.3. Starch Processing and Maltose Syrup Production

Example Recipe – High-Purity Maltose Syrup:

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

• Beta-Amylase: 100-500 g (depending on activity)

• pH: 5.0-5.5

• Temperature: 55-60°C

• Reaction time: 24-48 hours

Adjust liquefied starch to pH 5.0-5.5 and heat to 55-60°C. Add beta-amylase and incubate until maltose content reaches maximum (24-48 hours). Inactivate the enzyme by heat treatment and purify the syrup.

10.4. Other Applications

SECTION 11: ALTERNATIVES AND COMPARISON

11.1. Alpha-Amylase (EC 3.2.1.1)

11.2. Glucoamylase (Amyloglucosidase, EC 3.2.1.3)

11.3. Pullulanase (EC 3.2.1.41)

11.4. Maltogenic Alpha-Amylase (EC 3.2.1.133)

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 (P-Codes - 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 beta-amylase safe?
A1: Yes. Beta-amylase 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 beta-amylase and alpha-amylase?
A2: Alpha-amylase is an endo-enzyme that randomly cleaves the interior of the starch molecule. Beta-amylase is an exo-enzyme that systematically removes maltose units from the chain ends. This makes beta-amylase the preferred enzyme for high-purity maltose production.

Q3: Is beta-amylase suitable for vegan and halal products?
A3: Microbial (bacterial or fungal) or plant-derived beta-amylases are suitable for vegan and halal certification. There are no animal-derived beta-amylases. Relevant certificates can be obtained from manufacturers.

Q4: What is the shelf life of beta-amylase?
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 beta-amylase 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 beta-amylase?
A6: The optimum pH of beta-amylase is between 4.5-6.0, and the optimum temperature varies between 50-65°C depending on the source. Plant-derived enzymes typically show best activity around 55°C, while bacterial enzymes can remain active up to 60°C. The enzyme is rapidly inactivated above 70°C.

Q7: Which factors inhibit beta-amylase?
A7: Beta-amylase is inhibited by heavy metals (Cu²⁺, Hg²⁺, Ag⁺), SDS (Sodium Dodecyl Sulfate), cysteine, and β-cyclodextrin. Calcium (Ca²⁺) ions and some sulfhydryl compounds activate enzyme activity.

Q8: Why is beta-amylase important in beer production?
A8: In beer production, the natural beta-amylase in malt converts starch to fermentable sugars (especially maltose) during mashing. These sugars are then fermented by yeast to alcohol and CO₂. Beta-amylase activity directly affects the alcohol content, taste, and body of the beer.

Q9: How is high-maltose syrup produced with beta-amylase?
A9: For high-purity maltose syrup production, starch is first liquefied with alpha-amylase. Beta-amylase is then added and incubated at 55-60°C for 24-48 hours. This process converts approximately 50-60% of the starch to maltose. The enzyme is then inactivated by heat treatment and the syrup is purified.

Q10: What are the effects of inhaling beta-amylase powder?
A10: Beta-amylase 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 the baking industry 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: Beta-amylase 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: Beta-amylase is rapidly and irreversibly inactivated above 70°C. Strictly maintain application temperatures within the optimum range (50-65°C). Temperature control is critical in mashing or hydrolysis processes.

3. Activity Loss - pH: Beta-amylase loses stability below pH 4.0 or above pH 8.0. Maintain application pH within the optimum range (4.5-6.0). Excessively acidic or basic conditions cause permanent enzyme inactivation.

4. Activity Loss - Heavy Metals: Copper (Cu²⁺), mercury (Hg²⁺), and silver (Ag⁺) ions inhibit beta-amylase. 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 beta-amylase meeting specifications should be used in food applications. Industrial grade enzymes may contain impurities and are not suitable for human consumption.

6. Limit Dextrin: Beta-amylase cannot cleave the α-1,6 bonds at the branching points of amylopectin. When complete hydrolysis is required, it should be used in combination with a debranching enzyme such as pullulanase.

7. Natural Source: Plant-derived beta-amylases (barley, wheat) may contain gluten. For applications for celiac patients or those with gluten sensitivity, microbial (Bacillus, Aspergillus) beta-amylase should be preferred.

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.

• Enzyme Inactivation: Inactivate the enzyme by heat treatment (≥70°C, 10-15 minutes) 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.

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