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https://doi.org/10.15414/2019.9788055220703

4 International Scientific Conference Abstracts Book
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PROTEOLYTIC ACTIVITY OF MICROORGANISMS ISOLATED FROM BEES
AND WAX MOTHS
Tetiana Bulyhina, Oksana Brovarska

D.K. Zabolotny Institute of Microbiology and Virology of the National Academy of Science of Ukraine,
Kyiv, Ukraine; E-mail.: tati20@ukr.ner
Various types of bees have physical, physiological, behavioral, and chemical adaptations
which in addition to potential relationships with microbes and other organisms (wax moth),
may be particularly important for control spoilage of food stores rich in protein, lipid, and
carbohydrate content. It is known that Bacillus species isolated from provisions of bees
produced numerous extracellular enzymes involved in particularly in protein metabolism.
Protease activity has been commonly related to several functions, such as protein transport,
cellular, and tissue structuring, defense, and degradation/activation of proteins.
Therefore, the aim of this work was to study the ability of microorganisms isolated from
bees and wax moths to produce extracellular proteases.
To determine the enzymatic activity bacteria were grown on a liquid medium, the
following composition (g/l): KH2PO4 – 1.6; MgSo4 ·7H2O – 0.75; ZnSO4 ·7H2O – 0.25; (NH4)2SO4
– 0.5; maltose – 10.0; gelatin – 10.0; yeast autolysate – 0.15, pH – 6.5–6.7. Micromycetes were
grown on Chapek's medium (g/l): NaNO3 – 2.0; KH2PO4 – 1.0; KCl – 0.5; MgSO4 · 7H2O – 0.5;
FeSO4 · 7H2O – 0.015; gelatin – 20.0. The total proteolytic activity was carried out as a result of
casein hydrolysis. The elastase activity was determined colorimetrically by measuring the
change in the color intensity of the solution during enzymatic hydrolysis of elastin-congo red.
To establish the collagenase activity the optical density of the solution was measured after the
reaction of the collagen cleavage products with the ninhydrin reagent. Fibrinolytic activity
was identified after the formation of fibrin cleavage products.
Three fungi and one bacterium were isolated from bees, two fungi and one bacterium
from a wax moth, and one fungal and one bacterial culture from a cocoon of a wax moth.
Bacteria that were isolated from the samples (bees, wax moth and cocoons of the wax moth)
were very similar in character growth and morphological and cultural properties. As a result
of screening of 7 isolated microorganisms, only one fungal and one bacterial culture displayed
proteolytic activity, which was 0.01 and 0.0167 units/ml, respectively. Elastase activity
identified only in one fungal culture was 6.5 (when cultured in a test tube) and 12.3 units/ml
(when cultured in a flask). Fibrinolytic activity was 0.25 and 0,69 units/ml for bacterial and
fungal cultures, respectively. Collagenase activity was detected for both fungal and bacterial
culture (11.4 and 1.5 units/ml, respectively).
Our findings show that the fungal culture produces enzymes with elastolytic, fibrinolytic
and collagenolytic activity. In the future, fungal culture with elastolytic activity can be used in
bees pest control.

Keywords: proteolytic activity, bees, wax moths.

|76 4 International Scientific Conference Agrobiodiversity Nutrition, Health and Quality of Human and Bees Life
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September 11–13, 2019