This document discusses amylase, an enzyme that catalyzes the hydrolysis of starch into sugars. It begins by introducing amylase, noting its sources and industrial applications. It then classifies amylases and describes their mechanisms and effects. The document discusses microbial production methods, common producing strains, and materials and methods used. It outlines advantages and uses of microbial amylase and applications of specific amylases in industries like textiles, brewing, and syrup production.

1. School of Medical & Allied Sciences
Course Code: BPHT 6005 Course Name:Pharmaceutical Biotechnology
AMYLASE

2. INTRODUCTION
• Catalyses the hydrolysis of internal α-1,4-glycosidic linkages in starch in low molecular weight
products.
• Sources: Plants, Animals and Microorganisms.
• Large number of microbial amylases are available.
• Have almost completely replaced chemical hydrolysis of starch in starch processing industry.
• The amylases of microorganisms have a broad spectrum of industrial applications as they are more
stable than when prepared with plant and animal α-amylases.

3. • Amylase is an enzyme that catalyses the hydrolysis of starch into sugars.
• Present in the saliva of humans
• Hydrolysis of Starch with amylase will first result in the formation of a short polymer Dextrin and
then the disaccharide Maltose and finally glucose.
• Glucose is not as sweet as Fructose. Thus the next step would be the conversion of Glucose to
Fructose by the enzyme Glucose isomerase.

4. • Enzymes that break down starch or glycogen.
• Present in the saliva of humans and some other mammals, where it begins
the chemical process of digestion.
• All amylases are glycoside hydrolases and act on α-1,4-glycosidic bonds.
• Produced by a variety of living organisms, ranging from bacteria to plants
and humans.

5. CLASSIFICATION OF AMYLASES
α- Amylase
ß- Amylase
γ- Amylase

6. Glucose
Disaccharide maltose
Short chain polymer
Starch
Amylases
MECHANISM OF ACTION

7. • Also called as 1,4-α-D-glucan glucanohydrolase.
• Calcium metalloenzymes which cannot function in absence of calcium ions.
• Breaks down long carbohydrate chains of Amylose and Amylopectin.
• Amylose is broken down to yield maltotriose and Maltose molecules.
• Amylopectin is broken down to yield Limit dextrin and glucose molecules.
α- AMYLASE

8. • Found in saliva and pancreas.
• Found in plants, fungi (ascomycetes and basidiomycetes) and bacteria (Bacillus)
• Because it can act anywhere on the substrate, α-amylase tends to be faster-acting than β-amylase.
• In animals, it is a major digestive enzyme, and its optimum pH is 6.7–7.0

9. ß- AMYLASE
• Also called as 1,4-α-D-glucan maltohydrolase.
• Synthesized by bacteria, fungi, and plants.
• Working from the non-reducing end, β-amylase catalyzes the hydrolysis of the second α-1,4
glycosidic bond, cleaving off two glucose units (maltose) at a time.
• During the ripening of fruit, β-amylase breaks starch into maltose, resulting in the sweet flavor of
ripe fruit.
• The optimum pH for β-amylase is 4.0–5.0

10. • Also termed as Glucan 1,4-α-glucosidase.
• Cleaves α(1–6) glycosidic linkages, as well as the last α(1–4) glycosidic linkages at the nonreducing
end of amylose and amylopectin, yielding glucose.
• The γ-amylase has most acidic optimum pH of all amylases because it is most active around pH 3.
γ- AMYLASE

11. • Break down the starch
polymer but does not
give free sugar
Starch-
Liquefying
• Gives free sugars
Saccharogenic
EFFECTS OF α-AMYLASES

12. • Submerged fermentation (SmF)
• Solid state fermentation
Microbes utilized:
• Bacteria:
Bacillus licheniformis,
Bacillus stearothermophilus,
Bacillus subtilis,
Bacillus amyloliquefaciens
B. vulgarus,
• Fungi
Aspergillus and Penicillium
PRODUCTION METHODS AND MICROORGANISMS

13. • Bacteria – B. cereus, B.subtilis, B. amyloliquefaciens, B. polymyxa, B.
licheniformis etc
• Fungi – Aspergillus oryzae, Aspergillus niger, Penicillum, Cephalosporin,
Mucor, Candida etc.
PRODUCING STRAINS

14. • MICROORGANISM: Bacillus spp. was isolated from soil and maintained on nutrient agar slants and subcultured for
every 10 days.
• INOCULUM AND FERMENTATION MEDIUM: The inoculum was prepared by addition of sterile distilled water
into the freshly grown nutrient agar slants, from this 0.5ml of cell suspension was inoculated into 100ml of sterilized
fermentation medium and incubated at 35°C for 10 hrs.
• The composition of fermentation medium are:
Bacteriological peptone –6gm
Magnesium sulfate –0.5gm
Potassium chlorate –0.5gm
Starch –1gm (pH -7)
MATERIALS AND METHODS FOR PRODUCTION

15. EXTRACTION OF AMYLASE FROM FERMENTATION MEDIUM:
• After incubation the fermentation medium was harvested by centrifugation at 5000 rpm for 20 min at 4°C.The
supernatant was collected and subjected to estimate the amylase activity.
• EFFECT OF PH:
The fermentation medium was prepared by varying pH values(5.0, 6.0, 7.0, 8.0) for the production of amylase.
• EFFECT OF TEMPERATURE:
To study the effect of temperature on amylase production, the submerged fermentation was carried out at different
temperatures (25°C, 30°C, 35°C and 40°C).

16. • production of amylases is in economical bulk production capacity.
• microbes are also easy to manipulate to obtain enzymes of desired characteristics.
• The microbial amylases meet industrial demands a large number of them are available commercially.
• “BACILLUS LECHENIFORMIS”
“BACILLUS AMYLOLIQUIFACIENS”
“ASPERGILLUS NIGER”
• useful applications in food, brewing, textile, detergents and pharmaceutical industries.
• In detergents production, they are applied to improve cleaning effect and are also used for starch de-sizing in textile
industry.
• mainly employed for starch liquefaction (a process of dispersion of insoluble starch in aqueous solution) to reduce
their viscosity, production of maltose, oligosaccharide mixtures.
ADVANTAGES AND USES OF MICROBIAL AMYLASE

17. ENZYMES SOURCE APPLICATIONS
Alpha- amylase Bacterial α amylase (e.g., Bacillus
subtilis ), Fungal α amylase (e.g.,
Aspergillus niger )
Textiles, starch syrups, laundry and
dishwashing detergents, paper
desizing , fermentation
ß- amylase From a strain of Bacillus Brewing, maltose syrup
γ-Amylase Aspergillus niger Manufacture of dextrose syrup and
high fructose syrup production.
APPLICATIONS OF INDUSTRIALAMYLASES

18. Swati.Verma@galgotiasuniversity.edu.in