Mykola Kukhtyn, Khrystyna Kravcheniuk, Ludmila Beyko, Yulia Horiuk, Oleksandr Skliar, Serhii Kernychnyi


Microbial films formation on the dairy equipment creates a serious problem, because they are difficult to eliminate by washing and disinfecting means that results in contaminating dairy products by microorganisms. The aim of the work was to study the influence of Savinase®Evity 16L proteolytic enzyme on the process of destructing biofilms, formed by Staphylococcus aureus on stainless steel with different surface roughness.  

It has been established, that surface roughness of stainless steel influences the process of Savinase®Evity 16L enzyme penetration in a hollow and prevents the destruction of the biofilm matrix, created by Staphylococcus aureus.

It has been revealed, that after the influence of a proteolytic enzyme on Staphylococcus aureus biofilms, created on steel with roughness 0,16±0,018 mcm, the density decreased in 4,0 times (р≤0,05), comparing with a condition before processing. At roughness 0,63±0,087 mcm the density of formed biofilms decreased at the effect of Savinase®Evity 16L in 3,3times (р≤0,05) and the biofilm was characterized as a weak one. At the same time at stainless steel surfaces with roughness 2,68–0,95mcm, the density of biofilms decreased in 2,3–2,1times (р≤0,05), comparing with a condition before processing, and they were characterized as ones of the middle density. It has been also revealed, that the degradation intensity of biofilms under the influence of Savinase®Evity 16L enzyme at roughness 2,68–0,95 mcm was 1,7–1,9 times (р≤0,05) lower than at the surface with roughness 0,16±0,018 mcm.

So, the revealed degradation features of a biofilm, created by Staphylococcus aureus at surfaces of stainless steel of different roughness at the influence of Savinase®Evity 16L proteolytic enzyme give a possibility to substantiate the addition of proteolytic enzymes to the composition of washing means for dairy production. It is also offered to process the surface to the roughness no more than 0,63 mcm for producing food steel for raising the effectiveness of biofilms destruction by enzymes and for the sanitary processing.


enzyme Savinase®Evity 16L; destruction; density of biofilms; roughness of dairy equipment; sanitary processing

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Verran, J., Packer, A., Kelly, P., Whitehead, K. A. (2010). The retention of bacteria on hygienic surfaces presenting scratches of microbial dimensions. Letters in Applied Microbiology, 50 (3), 258–263. doi:

Kukhtyn, M., Berhilevych, O., Kravcheniuk, K., Shynkaruk, O., Horyuk, Y., Semaniuk, N. (2017). Formation of biofilms on dairy equipment and the influence of disinfectants on them. Eastern-European Journal of Enterprise Technologies, 5 (11 (89)), 26–33. doi:

Kukhtyn, M., Berhilevych, O., Kravcheniuk, K., Shynkaruk, O., Horyuk, Y., Semaniuk, N. (2017). The influence of disinfectants on microbial biofilms of dairy equipment. EUREKA: Life Sciences, 5, 11−17. doi:

Shaheen, R., Svensson, B., Andersson, M. A., Christiansson, A., Salkinoja-Salonen, M. (2010). Persistence strategies of Bacillus cereus spores isolated from dairy silo tanks. Food Microbiology, 27 (3), 347–355. doi:

Zhao, K., Tseng, B. S., Beckerman, B., Jin, F., Gibiansky, M. L., Harrison, J. J. et. al. (2013). Psl trails guide exploration and microcolony formation in Pseudomonas aeruginosa biofilms. Nature, 497 (7449), 388–391. doi:

Verran, J., Airey, P., Packer, A., Whitehead, K. A. (2008). Chapter 8 Microbial Retention on Open Food Contact Surfaces and Implications for Food Contamination. Advances in Applied Microbiology, 223–246. doi:

Langsrud, S., Moen, B., Møretrø, T., Løype, M., Heir, E. (2016). Microbial dynamics in mixed culture biofilms of bacteria surviving sanitation of conveyor belts in salmon-processing plants. Journal of Applied Microbiology, 120 (2), 366–378. doi:

Cherif-Antar, A., Moussa–Boudjemâa, B., Didouh, N., Medjahdi, K., Mayo, B., Flórez, A. B. (2015). Diversity and biofilm-forming capability of bacteria recovered from stainless steel pipes of a milk-processing dairy plant. Dairy Science & Technology, 96 (1), 27–38. doi:

Merritt, K., An, Y. H. (2000). Factors Influencing Bacterial Adhesion. Handbook of Bacterial Adhesion, 53–72. doi:

García, S., Trueba, A., Vega, L. M., Madariaga, E. (2016). Impact of the surface roughness of AISI 316L stainless steel on biofilm adhesion in a seawater-cooled tubular heat exchanger-condenser. Biofouling, 32 (10), 1185–1193. doi:

Marchand, S., De Block, J., De Jonghe, V., Coorevits, A., Heyndrickx, M., Herman, L. (2012). Biofilm Formation in Milk Production and Processing Environments; Influence on Milk Quality and Safety. Comprehensive Reviews in Food Science and Food Safety, 11 (2), 133–147. doi:

Cloete, T. E. (2003). Resistance mechanisms of bacteria to antimicrobial compounds. International Biodeterioration & Biodegradation, 51 (4), 277–282. doi:

Davin-Regli, A., Pages, J. M. (2012). Cross-resistance between biocides and antimicrobials: an emerging question. Revue Scientifique et Technique de l’OIE, 31 (1), 89–104. doi:

Simões, M., Simões, L. C., Vieira, M. J. (2010). A review of current and emergent biofilm control strategies. LWT – Food Science and Technology, 43 (4), 573–583. doi:

Oliveira, N. M., Martinez-Garcia, E., Xavier, J., Durham, W. M., Kolter, R., Kim, W., Foster, K. R. (2015). Correction: Biofilm Formation As a Response to Ecological Competition. PLOS Biology, 13 (8), e1002232. doi:

Monds, R. D., O’Toole, G. A. (2009). The developmental model of microbial biofilms: ten years of a paradigm up for review. Trends in Microbiology, 17 (2), 73–87. doi:

Lopez, D., Vlamakis, H., Kolter, R. (2010). Biofilms. Cold Spring Harbor Perspectives in Biology, 2 (7), a000398–a000398. doi:

Römling, U., Kjelleberg, S., Normark, S., Nyman, L., Uhlin, B. E., Åkerlund, B. (2014). Microbial biofilm formation: a need to act. Journal of Internal Medicine, 276 (2), 98–110. doi:

Krushelnytska, N. V. (2013). The influence of PH medium on the ability to form microbial biological tape by microorganisms extracted from milking equipment and raw milk. Naukovo-tekhnichnyi biuleten Instytutu biolohiyi tvaryn i Derzhavnoho naukovo-doslidnoho kontrolnoho instytutu vetpreparativ ta kormovykh dobavok, 14 (3-4), 82–86. Available at:

Lequette, Y., Boels, G., Clarisse, M., Faille, C. (2010). Using enzymes to remove biofilms of bacterial isolates sampled in the food-industry. Biofouling, 26 (4), 421–431. doi:

Sandhya, C., Sumantha, A., Pandey, A. (2004). Proteases. Enzyme Technology. Asiatech Publishers Inc., New Delhi, 312–325.



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