Submerged culture fermentation for production of amylase and down-stream processing


Submerged fermentation is a type of fermentation where the organism can grow beneath the surface of the medium. Submerged culture fermentation has been widely used for production of enzyme because in submerged fermentation unwanted metabolites are not produced and purification of enzymes takes place in easy way.


  1. Aspergillus niger or Bacillus culture.
  2. Soluble starch
  3. Peptone
  4. MgSO4
  5. KCl
  6. Glucose
  7. Lactose
  8. Mannitol
  9. Sucrose
  10. Urea
  11. NH4Cl
  12. Tween 80
  13. 0.1mM acetate Buffer
    14. DNS reagent
    15. Whatman filter paper no. 1


  1. Submerged fermentation was carried out in the Erlenmeyer flasks by taking 100 mlof amylase production medium containing containing Peptone (6.0g/L), MgSO4(0.5g/L), KCl (0.5g/L), Starch (1g/L), Carbonsources (each 2g/L) included are glucose, mannitol, starch,sucrose, lactose. Nitrogen sources included are NH4Cl, NaNO3,KNO3, Peptone, Urea and Yeast extract (2g/L) as nitrogen source.
  2. Make primary inoculum by inoculating 10 ml of the above media with the given culture. Keep it overnight at 30°C on a shaker with 120rpm.
  3. Using the primary inoculum, make secondary inoculum by transferring 5% of the primary inoculum to fresh media. Keep it overnight at 30°C on a shaker with 120rpm.
  4. Transfer 5% of the secondary inoculum to the production media.
  5. The medium was maintained at a pH range of 7 at 30°C on a shaker with 120rpm for 6 to 18 days.
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