Enzyme preparation of microbial origin and evaluation of its effectiveness in the processing of collagen-containing raw materials

S. L. Tikhonov¹, I. S. Brashko¹, N. V. Tikhonova¹, M. S. Tikhonova^2
1Ural State University of Economics, ul. 8 Marta/Narodnoi Voli, 62/45, Ekaterinburg, 620144, Russian Federation
²Ural State Medical University, ul. Repina, 3, Ekaterinburg, 620028, Russian Federation

Abstract. The purpose of the studies was to obtain an enzyme preparation based on collagenase of microbial origin and to assess its proteolytic properties. For this purpose, Cl. histolyticum was cultured on a nutrient medium containing phytopeptone from lupine seeds followed by the separation of the metabolic product, i.e. a collagenase enzyme, from the nutrient medium by ultrafiltration on membrane transmitting macromolecules weighing up to 130 kDa. The degree of hydrolysis of the lupine protein in phytopeptone was determined by the content of amine nitrogen. The proteolytic activity of the enzyme preparation was assessed by hydrolysis of collagen-containing raw materials – crushed broiler chicken skin. For the experiment, we formed two batches of samples of 5 kg each. The first was the control one; the samples from the second batch were treated with an enzyme preparation in the amount of 0.3% of the raw material mass for 8 hours. The amine nitrogen content in the crushed skin hydrolyzate increased strongly in the first hours. In 2, 3, 4, and 5 hours of hydrolysis, it was 2.9 g/L, 3.8 g/L, 4.5 g/L, and 5.2 g/L, respectively. These indicators were higher than in control samples 14.5, 19.0, 22.5, and 26.0 times respectively. The enzyme preparation was characterized by a high proteolytic activity against collagen-containing raw materials, as evidenced by the high content of amine nitrogen (5.4 g/L), nitrogen of sarcoplasmic and myofibrillar proteins (15.9% and 2.7%, respectively), non-protein and polypeptide nitrogen (8.1% and 3.3%, respectively) to the total hydrolyzate nitrogen after 6 hours of hydrolysis, which was 27, 41, 14, 82, and 66 times higher than in the samples of crushed skin from the control batch. Histological examination of collagen-containing raw materials against the background of the introduction of the enzyme showed loosening of the fibres and their homogenization with the formation of cavities of the protein mass.

Keywords: nutrient medium; cultivation; microorganisms; hydrolysis; collagen-containing raw materials; enzymes.

Author Details: S. L. Tikhonov, D. Sc. (Tech.), prof. (Этот адрес электронной почты защищён от спам-ботов. У вас должен быть включен JavaScript для просмотра.); I. S. Brashko, post graduate student; N. V. Tikhonova, D. Sc. (Tech.), prof.; M. S. Tikhonova, student.

For citation: Tikhonov SL, Brashko IS, Tikhonova NV, et al. [Enzyme preparation of microbial origin and evaluation of its effectiveness in the processing of collagen-containing raw materials] Dostizheniya nauki i tekhniki APK. 2021;35(9):67-72. Russian. doi: 10.53859/02352451_2021_35_9_67.