Amylase Production on
by Bacillus spp.
Miss Karishma Gangwani
MSc. Biotechnology sem II
Hislop School Of
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-glycosidic bonds.
produced by a variety of living organisms, ranging from bacteria to plants
Classification Of Amylases
based on how they break down starch molecules :
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
The microbial amylases meet industrial demands a large number of
them are available commercially.
useful applications in food, brewing, textile, detergents and
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.
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)
Extraction Of Amylase From Fermentation
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).
Assay of Amylase:
The amylase activity is determined following the method of
Bernfeld. An assay mixture containing, enzyme extract, starch as a
substrate and DNSA as coupling reagent was used.
One unit of amylase activity was defined as the number of micro
moles of maltose liberated by 1ml of enzyme solution per minute.
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,
desizing , fermentation
ß- amylase From a strain of
γ-Amylase Aspergillus niger Manufacture of
dextrose syrup and
high fructose syrup