Isolation and screening of protease producers

/Isolation and screening of protease producers
Isolation and screening of protease producers 2018-02-23T10:55:35+00:00

Isolation and screening of protease producers

Isolation and screening of protease producers.

Introduction

Protease refers to a group of enzymes whose catalytic function is to hydrolyze (breakdown) peptide bonds of proteins. They are also called proteolytic enzymes or proteinases. Proteases differ in their ability to hydrolyze various peptide bonds. Each type of protease has a specific kind of peptide bonds it breaks. Examples of proteases include: fungal protease, pepsin, trypsin, chymotrypsin, papain, bromelain, and subtilisin. Proteases occur naturally in all organisms. These enzymes are involved in a multitude of physiological reactions from simple digestion of food proteins to highly regulated cascades (e.g., the blood-clotting cascade, the complement system, apoptosis pathways, and the invertebrate prophenoloxidase-activating cascade). Proteases can either break specific peptide bonds (limited proteolysis), depending on the amino acid sequence of a protein, or break down a complete peptide to amino acids (unlimited proteolysis). The activity can be a destructive change, abolishing a protein’s function or digesting it to its principal components; it can be an activation of a function, or it can be a signal in a signaling pathway. Proteolytic enzymes are very important in digestion as they breakdown the protein foods to liberate the amino acids needed by the body. Additionally, proteolytic enzymes have been used for a long time in various forms of therapy. Their use in medicine is gaining more and more attention as several clinical studies are indicating their benefits in oncology, inflammatory conditions, blood rheology control, and immune regulation.

Requirements

  1. Soil samples
  2. Distilled water
  3. Nutrient agar
  4. Petriplates
  5. Skimmed milk powder
  6. Toothpicks

Procedure

  1. Obtain soil samples and make serial dilutions.
  2. Spread 0.1ml of an appropriate dilution on a nutrient agar plate. Incubate overnight at 37°C.
  3. Now separate the colonies and streak them on nutrient agar plates to obtain pure cultures.
  4. Make nutrient agar plates enriched with 1% skimmed milk powder. Streak these plates with the pure cultures isolated and incubate overnight.
  5. Check for growth and zone of clearance around the colonies. A clearance zone indicates the presence of proteases.
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