Mikrobiol. Z. 2020; 82(4):53-62. Ukrainian.
doi: https://doi.org/10.15407/microbiolj82.04.053

The Influence of Cultivation Parameters on Penicillium restrictum
α-L-Rhamnosidase Activity

O.V. Gudzenko, N.V. Borzova, L.D. Varbanets

Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

The efficiency of the process of obtaining biotechnologically active substances, in particular enzymes, by microbiological synthesis largely depends on the successful optimization of the conditions of producer cultivation. The aim was to determine the influence of the main components of nutrient medium and culture conditions on the level of extracellular α-L-rhamnosidase activity of Penicillium restrictum. Methods. The micromycetes culture was grown in a submerged cultivation condition at 25°C for 7 days. α-L-Rhamnosidase activity in the supernatant culture liquid of P. restrictum was determined using n-nitrophenyl-α-L-rhamnopyranoside. Xylose, arabinose, glucose, galactose, rhamnose, mannose, lactose, maltose, sucrose, and mannitol were used as carbon sources. Sodium nitrate, sodium nitrite, ammonium sulfate, ammonium nitrate, yeast autolysate, yeast extract, peptone, urea, and soy flour were used as nitrogen sources. To study the effect of cultivation conditions, the strain was grown on medium optimized for carbon and nitrogen sources, changing the initial pH of the medium (3.0–8.0), temperature (25–42°C) and volume of the medium (50–250 ml). Results. It was found that the highest rates of P. restrictum α-Lrhamnosidase activity were observed on the 7th day of cultivation in case of rhamnose using a as carbon source at a concentration of 5 g/l and ammonium sulfate as nitrogen source – 0.8 g/l. The most effective for α-L-rhamnosidase synthesis by P. restrictum was the use a medium with an initial pH value of 6.0, temperature of 25°C and 100 ml of nutrient medium in flasks. Conclusions. It was shown the possibility of P. restrictum to synthesize a new highly active α-Lrhamnosidase in submerged cultivation conditions. The combination of rhamnose and ammonium sulfate was found to provide α-rhamnosidase activity (1.2 U/ml). The established physical and chemical conditions of P. restrictum cultivation can be used for production of α-L-rhamnosidase for biotechnological purposes.

Keywords: α-L-rhamnosidase, Penicillium restrictum, cultivation parameters, carbon and nitrogen sources.

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