Mikrobiol. Z. 2021; 83(2):12-19.
Screening of Enzyme Producers with Keratinase Activity among Marine Actinobacteria
K.V. Avdiyuk1, V.A. Ivanytsia2, L.D. Varbanets1
1Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
2Odessa Mechnikov National University
2 Dvoryanskaya Str., Odessa, 65029, Ukraine
About 2 million tons of feathers are produced annually around the world as a by-product of poultry farming. Due to the lack of funds and the complexity of processing, they have become one of the main environmental pollutants. The biodegradation of feathers by keratinolytic microorganisms has proven to be an effective, environmentally friendly and cost-effective method of bioconverting feather waste into a nutritious, balanced and easily digestible product that contains free amino acids, peptides and ammonium ions. Aim. To investigate the ability of marine actinobacteria to synthesize enzymes with keratinolytic activity and to study some of the physicochemical properties of the most active enzyme preparation. The object of the study was 10 strains of actinobacteria isolated from bottom sediments in the area of the Pradneprovsky trench of the Black Sea shelf. Methods. Caseinolytic (general proteolytic) activity was determined by the Anson method modified by Petrova, based on the quantitative determination of tyrosine, which is formed during the enzymatic hydrolysis of casein. Keratinase activity was determined by UV absorption at 280 nm of the hydrolysis products of keratin-containing raw materials. The cultivation of actinobacteria was carried out in a liquid nutrient medium with the addition of defatted chicken feathers as the main source of carbon and nitrogen. Results. The ability to hydrolyze keratin was found in five cultures. Moreover, all the strains studied were practically unable to break down casein. The Acty 9 strain (12 U/ml) showed the highest keratinase activity. Additional introduction of NaCl to the nutrient medium did not have a positive effect on the enzymes synthesis. The study of the physicochemical properties of the enzyme preparation Acty 9 showed that the pH and thermooptimum were 9.0 and 60°C, respectively. It retained 100% of the initial activity in the range of pH 7.0–10.0 after 3 h and 95% activity at pH 8.0 after 24 h of incubation. The studied enzyme preparation was thermostable, since it remained active for 3 h at 50°C and 1 h at 60°C. Conclusions. The extracellular keratinase synthesized by actinobacterium Acty 9 is promising for further research, since the enzyme is pH and thermostable and is not inferior in its physicochemical properties to those previously described in the literature.
Keywords: actinobacteria, keratinolytic enzymes, producers, caseinolytic activity.
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