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An Enzymatic Process for Debittering of Protein Hydrolysates using Immobilized Peptidases


Protein hydrolysis is widely practiced in food industry for variety of purposes like nutritional supplement. They are also gaining acceptance as components of sports and weight control diets. One of the attributes of protein hydrolysates that negatively influence their consumer acceptance is bitterness. Chicken intestine is a rich source of diverse aminopeptidases, which are largely (70-89%), located in the mucosa. Using chicken intestinal mucosa as the source of peptidases, a novel method for the enzymatic debittering of the protein hydrolysates has been developed after rendering it free of micro flora by gamma irradiation (20 kGy) but retaining the enzymatic activity. The radiation hygienised mucosa was entrapped in calcium alginate beads. The immobilized beads exhibited aminopeptidase activities. Hydrolysis of hydrophobic residues by exopeptidases could lead to reduction in bitterness. Thus, when bitter protein hydrolysate is passed over the column containing these beads, bitter peptides are cleaved, resulting in debittered protein hydrolysate, which comes out from another end. Use of immobilized enzymes ensures use of the column for continuous debittering of protein hydrolysates. 

Protein hydrolysis is widely practiced in food industry for variety of purposes which includes, improvement of nutritional characteristics, retarding deterioration, modification of functional properties and the removal of toxic or inhibitory ingredients. Protein hydrolysates also form an important part of medical diets for the treatment of short bowel syndrome, Crohn?s disease and diets for elderly. They are also gaining acceptance as components of sports and weight control diets. One of the attributes of protein hydrolysates that negatively influence their consumer acceptance is bitterness which has been assigned to low molecular weight (> 10 kDa) peptides containing pro, leu, tyr, phe, ala etc in specific combinations.

 ADVANTAGES

  • The process involves use of exopeptidases from poultry intestine, a processing byproduct, available abundantly at very low cost.

APPLICATIONS

This technology has applications in following areas:

  • Protein food industries.
  • Pharmaceutical industries.

Protein hydrolysis is widely practiced in food industry for variety of purposes which includes, improvement of nutritional characteristics, retarding deterioration, modification of functional properties and the removal of toxic or inhibitory ingredients. Protein hydrolysates also form an important part of medical diets for the treatment of short bowel syndrome, Crohn?s disease and diets for elderly. They are also gaining acceptance as components of sports and weight control diets. One of the attributes of protein hydrolysates that negatively influence their consumer acceptance is bitterness which has been assigned to low molecular weight (> 10 kDa) peptides containing pro, leu, tyr, phe, ala etc in specific combinations. Debittering of the hydrolysate would thus make product better acceptable by consumers.

WORKING PRINCIPLE

Chicken intestine is a rich source of diverse aminopeptidases, which are largely (70 - 89 %) located in the mucosa. Using chicken intestinal mucosa as the source of peptidases, a novel method for the enzymatic debittering of the protein hydrolysates has been developed after rendering it free of the micro flora by gamma irradiation (20 kGy). Irradiation is a cold sterilization process that hygienises the tissue with least effect on proteolytic enzymes. The radiation hygienised mucosa was entrapped in calcium alginate beads, which is accepted as a food grade system. The immobilized beads exhibited aminopeptidase activities with preference to neutral and hydrophobic N terminal amino acids, which incidentally are responsible for the strong bitterness of peptides. Hydrolysis of hydrophobic residues by exopeptidases could lead to reduction in bitterness. Thus, when bitter protein hydrolysate is passed over the column containing these beads, bitter peptides are cleaved, resulting in debittered protein hydrolysate, which comes out from another end. Use of immobilized enzymes ensures use of the column for continuous debittering of protein hydrolysates.

PRODUCT CHARACTERIZATION

  • The product has been characterized for amino acid profile, peptide chain length, functional properties, sensory analysis and RP HPLC (Profile is an indicator of debittering process).

ADVANTAGES

  • The process involves use of exopeptidases from poultry intestine, a processing byproduct, available abundantly at very low cost.

APPLICATIONS

This technology has applications in following areas:

  • Protein food industries.
  • Pharmaceutical industries.

REQUIREMENTS

Infrastructure

  • Washing room: Small room (10? X 10?) with water connection and tap fitting, for initial storage and superficial cleaning of the intestinal tissue.
  • Walk in cooler: set at 40C temperature (size: 8? X 8? X 7.5?)
  • Gamma Chamber 5000: Setting up this facility is optional as radiation processing of the mucosa could be outsourced.
  • A small room (10? X 10?) for preparation of beads.
  • A Small room (10? X 10?) with water and electricity connection for setting up column containing immobilized beads.

Equipment required

  • Peristaltic Pump
  • A glass column
  • Weighing Balance
  • Column holders
  • A circulating water bath

Statutory requirements

  • The registration/ license as per FSSAI guidelines.

Manpower requirement

  • For 6 kg manual processing daily, 1 unskilled and 1 skilled worker are required.


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General License Fee : ₹ 71,000.00 (Seventy One Thousand)

General Royalty : 0%