Amylase production

1. Amylase Production
Lasina .j
Maris Stella College

2. What Are Amylases?
 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-glycosidicbonds.
 produced by a variety of living organisms, ranging from bacteria toplants
and humans.

3. Classification Of
Amylases
 based on how they break down starch molecules :
1. α-Amylase
2. β-Amylase
3. γ-Amylase

4. Advantages And Uses Of Microbial Amylase
 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”
“BACILLUSAMYLOLIQUIFACIENS”
“ASPERGILLUS NIGER”
 useful applications in food, brewing, textile, detergentsand
pharmaceutical industries.
 In detergents production, they are applied to improve cleaningeffect
and are also used for starch de-sizing in textileindustry.
 mainly employed for starch liquefaction (a process of dispersion of
insoluble starch in aqueous solution) to reduce their viscosity,
production of maltose, oligosaccharide mixtures.

5. Materials And Methods For Production
 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)

6. 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 supernatantwas
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 OFTEMPERATURE:
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).

7. Assay of Amylase:
 The amylase activity is determined
following the method of Bernfeld. An
assay mixture containing, enzyme
extract, starch asa substrate and DNSA
as coupling reagent wasused.
 One unit of amylase activity was
defined as the number of micro moles
of maltose liberated by 1ml of enzyme
solution per minute.

8. Applications Of Industrial Amylases:
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.

9. THANK YOU!!