Production technology and biocatalytic properties of proteolytic enzyme preparation

The purpose of the study is to develop a technology for obtaining a proteolytic enzyme preparation based on collagenase isolated from the entrails of fish and to study the biocatalytic activity depending on pH and temperature. The proteolytic activity of the enzyme preparation was determined by the Anson method. A technology for producing an enzyme preparation has been developed, which includes isolation of chum salmon entrails, processing of fish raw materials with an ultra-high pressure of 100-200 MPa for 60 seconds, grinding to particles of 500-600 microns, extraction and homogenization in an isotonic solution in a ratio of 1:3 at a temperature of 35-37 °C for 3-4 hours with a rotation frequency of 100-200 revolutions per minute, centrifugation (3000 rpm); ultrafiltration through ceramic membranes with a transmission limit of up to 21 kDa, packaging in plastic bags with a capacity of 50 ml, sterilized with an ultra-high pressure of 300 MPa for 60-120 seconds. The activity of the enzyme preparation was determined by the express method, which consists in hydrolysis of gelatin with a collagenase solution. As a result of the study, a technology for the production of an enzyme preparation from fish raw materials treated with an ultrahigh pressure of 100-200 MPa for 60 seconds was developed. The enzyme preparation was controlled using a temperature regime in order to prevent inactivation of the enzyme. Extraction and homogenization were carried out in an isotonic solution in a ratio of 1:3 at a temperature of 35-37 °C for 3-4 hours with a blade rotation speed of 100-200 revolutions per minute. The temperature during extraction was set using a control unit that regulates the operation of the compressor, which is part of a complex synchronous mixing unit.The applied technology of ultrafiltration of homogenizate consists in the separation of the initial solution supplied to the circulation tank into an enzyme preparation and an aqueous solution continuously discharged from the apparatus. When producing an enzyme preparation using this technology, in order to prevent the inactivation of the enzyme, the temperature regime during homogenization was monitored by setting the necessary temperature that regulates the operation of the compressor with the cooling liquid.During ultrafiltration of the temperature control homogenizate, a classic Dewar vessel in the form of a sealed foam container with ice placed inside was used. The influence of pH and temperature on the biocatalytic activity of the enzyme preparation was established in the course of the studies. optimum of the activity of the samples of the enzyme preparation-observed at a pH of 7 to 9 of the activity of the enzyme preparation, temperature. The pH shift to the alkaline side leads to a decrease in activity. It was found that the optimum of the catalytic activity of the enzyme preparation is at a temperature range of 37 to 42 °C.