Influence of Priestia megaterium 035 Culture Parameters on Keratinase Synthesis
DOI:
https://doi.org/10.15407/Keywords:
Priestia megaterium,, keratinase,, parameters of cultivationAbstract
The accumulation of a huge amount of keratin waste as by-products of poultry and livestock farming has become a global problem today. This problem can be solved by using keratinolytic enzymes that are able to break down poorly soluble keratin proteins, thereby reducing the negative anthropogenic effect on the environment. The aim of this work was to investigate the effect of various parameters and cultivation conditions on keratinase synthesis by Priestia megaterium 035 culture. Methods. P. megaterium 035 culture was grown under submerged cultivation conditions in 750 mL Erlenmeyer flasks at a temperature of 28 °C on a basic nutrient medium containing defatted chicken feathers as the only source of carbon and nitrogen for 3–5 days. The study of the influence of various parameters and cultivation conditions on keratinase synthesis by P. megaterium 035 culture was carried out according to the following criteria: the use of different carbon and nitrogen sources, substrate concentrations, cultivation temperatures, shaker rotation speed, volume of the nutrient medium, and initial pH values. Protein was determined by the Lowry method, keratinase activity was estimated by light absorption by hydrolysis products of keratin-containing materials in the ultraviolet part of the spectrum. Results. The study of the effect of additional carbon sources on keratinase activity of P. megaterium 035 showed that its highest level was observed on a medium with maltose; slightly lower results were obtained on a medium with galactose. The addition of glucose inhibited the synthesis of keratinase P. megaterium 035. The introduction of additional nitrogen sources into the nutrient medium showed that the use of bactotryptone and soybean flour increased the synthesis of keratinase P. megaterium 035 by 1.6 and 1.25 times, respectively. Complete inhibition of enzyme synthesis was observed when inorganic nitrogen sources (ammonium nitrate and sulfate) were added. The study of the influence of some physicochemical parameters on the keratinase activity of P. megaterium 035 showed that the maximum value of enzymatic activity was observed at pH 10.0, temperature 40 °C, stirring intensity 200 rpm, nutrient medium volume 50 mL, substrate concentration 0.75 g/L, and inoculum amount 15%. Conclusions. Selection of optimal conditions and cultivation parameters made it possible to increase the keratinase activity of P. megaterium 035 by twice compared to the original medium and significantly reduce the time of feather splitting from three to one and a half days.
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