INDUSTRIAL PRODUCTION OF AMYLASE FROM FUNGUS

AMYLASE, TYPES,METHODS,APPLICATIONS

1. Industrial Production Of Amylase From Fungus Shraddha Mahadeorao Madghe B.sc 3 rd year 6 th sem {MBZ} Department of Microbiology Guided By
Dr. C. S.Ukesh Mam Dr. Ashruta Gawali Mam

2. Contents Introduction
Sources Of Enzymes
Amylase Types Of Amylase Methods Of Manufacturing Production Of Amylase By Surface Culture Method
Industrial Applications

3. Introduction Enzymes are proteins hence posses physical characteristics of proteins. Each enzyme has its own pH and temperature for optimum activity. Enzyme show specificity for the particular substrate that they attack for example amylase acts on starch and protease on proteins.

4. Sources of Enzyme Since all the living cells produce enzyme it is possible to obtain useful enzymes from : 1. Animal tissue : Not considered A significant source for the commercial production of amylase . Agricultural wastes are used in the production. 2 . Plant tissue :. Ptyalin, A salivary a-amylase (40% total salivary protein) calcium containing metalloenzyme , hydrolyzes Α -1,4 linkages of starch.
3. Microorganism : Fungal ( Aspergillus niger , Aspergillus oryzae, Penicillium , thermomyces , Lanuginosus ), bacterial (bacillus spp.)

5. Amylase Amylase is an enzyme that catalyses the hydrolysis of starch into sugars.
Amylase is present in the saliva of humans and some other mammals, where it begins the chemical process of digestion.
These enzymes randomly cleave internal glycosidic linkages in starch molecules to hydrolyze them and yield dextrins and oligosaccharides.

6. Types of Amylase All amylases are glycoside hydrolyses and act on a-1,4-glycosidic bonds
Amylase can be classified into 3 types: 1. α - Amylase: .2. β -Amylase 3. γ -Amylase

7. General Methods Of Manufacture Surface Culture Method : In this method the organisms are grown on the surface of the solid, semi solid or liquid medium. Submerged Culture Method : In this method the organisms are grown in submerged condition in a liquid medium.

8. Production Of Fungal Amylase By The Surface Culture Method Organism Selected Inoculum Preparation Medium Preparation Sterilization And Inoculation Incubation
Recovery

9. Organism selected Amylase is the extracellular enzyme which hydrolysis the starch. Starch glucose maltose
Selected strains of Aspergillus oryzae (ATCC 4814) is selected for amylase production by Koji process. The stock culture is maintained as a pure culture on Czapek agar or Wort agar

10. Inoculum preparation Spores of Aspergillus oryzae are used as inoculum for the enzyme production spores are prepared in a small seed bottle rather than in a large bottle The medium for small seed bottle is prepared by mixing 15 grams of wheat bran and 12 ml of water. Small seat bottle are inoculated with the spore material from agar slant of Aspergillus oryzae . The medium for large seed bottle is the mixture of the following ingredients : wheat bran : 100 grams peptone : 2.5 grams water : 110 ml In a large sheet bottle 400 grams of this mixture is placed in the opening is closed by gauge. Bottles are sterilized for 30 minutes at 120°C on the three successive days. Each bottle is then inoculated with a small quantity of dry spore material from small seat bottle and incubated at 30°C until the material is heavily grown sporulated and dried.

11. Medium Preparation Sterilization And Inoculation Medium is prepared in large rotary drum. Appropriate quantity of wheat bran and water added in a rotary drum and it is sterilized by direct streaming at a pressure of 15 LBS per inch for 3 hours
After that filtered cool air is blown through drum, cold water is also circulated over the drum, until mixture is cooled to 31°C. Then mixture is inoculated by adding dry spore culture for 20 large bottles and rotating drum for 1 hour to ensure uniform distribution of inoculum.

12. Incubation The inoculated brand sprayed on the perforated trays in a layer of about 1 inch deep and trays are then placed into the incubation chamber. The temperature of air is raised to 37°C to initiate the growth and during active growth period temperature is maintained between 38°C to 41°C. During incubation period mold mycelium growth throughout the bran mass and forms Koji. Maximum amylase potency is obtained after incubation for 24 hours to 36 hours.

13. Recovery The enzyme is extracted into water by passing water through Koji, then pumping the solution from the bottom of the first percolator to the top of the next circulator in the series and so on. The first 50 gallons of the concentrated extract is processed for the for enzyme recovery and 2 nd 50 gallons of the week extract is used to study the extraction of the next batch of the Koji. Enzyme can be precipitate from the extract by adding cold 95% special denatured alcohol agitating slowly for about an hour and then allow the precipitate to settle. The clear supernant liquid is decanted off. Precipitate slurry and equal volume of alcohol is mixed in the tank. Enzyme precipitate was filtered and dried at 41°C in an atmospheric dryer. The drains and concentrate is removed from the dryer and ground to a fine powder and transfer to a moisture proof container. The enzyme concentrate is sampled and assayed in laboratory for amylase activity to know it’s potency.

14. Applications Hydrolysis of starch into the extreme forming a less viscous starch suspension.
In bakers industry, converting starch in two smaller fermentable sugars.
In textile industry, used to removal of starch sizing agent for woven fabric. Fuel alcohol production, converting starch into smaller fermentable.
Sugars which are acted upon by east to produce alcohol.

15. Thank you !