Laboratory of Gene Engineering and Biotechnology of Lipophilic Bioactive Compounds

Faculty of Chemical and Food Technology

Department/Institute: Department of Biochemical Technology
Contact person:         Assoc. Prof. Milan Čertík, PhD.
Contact:                   +421 917 669 410
e-mail:                      milan.certik@stuba.sk
web page:                 http://www.fchpt.stuba.sk/generate_page.php?page_id=2218

Description:

The laboratory of Assoc. Prof. Milan Čertík is focusing on study of microbial lipids and pigment biosynthesis as well as on biotechnological regulation and overproduction of biologically active metabolites such as polyunsaturated fatty acids, pigments and glycoproteins. The laboratory is also focused on gene engineering and characterization of the main genes coding biosynthesis of biologically active lipids and carotenoid pigments and their functional expression into microorganisms and plants. The team employs various microbial strains for submerged fermentations (zygomycetes filamentous fungi, “red” yeasts, oleaginous yeasts) and solid state fermentations (zygomycetes filamentous fungi) to reach the overproduction of desired metabolites and to describe the metabolic key steps involved to biosynthesis of lipid structures ad pigments. Especially fungal solid state fermentations resulted in several cereal-based bioproducts functionally enriched with polyunsaturated fatty acids and pigments. These bioproducts have been tested as a food and feed additives. The laboratory is furnished with equipments for various types of microbial cultivations, for gene engineering techniques as well as for detailed analysis of studied metabolites. Scientific level of the team is given by its respected world-known experiences as well as by high publication activity and invited lectures in prestigious congresses. The laboratory consists of two employees (Assoc. Prof. Milan Čertík, PhD, Tatiana Klempová, PhD.), PhD, MSc and Bc students. The laboratory is internationally accepted and actively collaborates with several domestic and foreign Universities and Institutions.

 

Projects and publications:

Significant projects for last years

1. Improvement of nutrition value of various cereals by polyunsaturated fatty acids. National Research and Development Project “Quality, safety and functionality of primary food resources” No. 2003 SP 27/028 0E 02/028 0E 02. Ministerstvo of Agriculture SR, 2003-2005  
2. Evaluation of agroindustrial materials using biotechnological means by application of microorganisms producing metabolites for food, agriculture, pharmacy and veterinary uses. Project VEGA No. 1/2390/05, Ministry of Education SR, 2005-2007
3. Biotechnological production of bioactive lipids from agro-industrial byproducts. Bilateral scientific & technological project between the Slovak Republic and the Hellenic Republic, Gr/Slov/FCHPT/05, Ministry of Education SR, 2005-2006
4. Regulation of biotechnological production of conjugated fatty acids (CLA). National Research and Development Project No. 2003 SP 270280 E01 028 0E01. Ministry of Industry SR, 2003-2005
5. Phytosterols and fatty acid derivatives. National Research and Development Project No. 2004 ŠP 26 028 0C 05 „Utilization of national resources: Complex utilization of plant resources“. Ministry of Industry SR, 2004-2006
6. Biotechnological evaluation of national renewable resources to biologically active products applied for food, agriculture, pharmacy and veterinary uses. Project VEGA, Ministry of Education SR, 2008-2010
7. Yeast as a tool for producing biotechnologically valuable sterols: the biochemical and genetic approach. Project APVV-0681-07, 2008-2010
8. Biomembranes: Membrane structure and dynamics in relation to cell functions, Project of excellence VVCE-0064-07, 2008-2011
9. Investigations of factors affecting oxidation of frying oils and fried products. International Project with PepsiCo International (USA), 2008 – 2010
10. Vybudovanie kompetenčného centra pre výskum a vývoj v oblasti molekulárnej medicíny. Projekt Štrukturálne fondy EU, Operačný program Výskum a Vývoj, ITMS projektu: 26240220071, 2011-2014
11. Biotechnological evaluation of domestic agricultural resources for bioproducts with aimed nutritional and functional properties. Project VEGA No. 1-0975-12, Ministry of Education SR, 2012-2014
12. Scale-up of solid state fermentation process for polyunsaturated fatty acids production by fungi. National fellowship program – SAIA, Thailand, 2012
13. Investigations of antioxidant compounds preventing oxidation of frying oils and fried products. International Project with PepsiCo International (USA),, 2011-2012
14. Biotechnological preparation of new types of functional cereals and cereal products enriched with polyunsaturated fatty acid and pigments, Project APVV 0662-11, 2012 – 2015
15. Nonconvetional approach of preparation of cereals with enhanced economical potencial, Project APVV-0294-11, 2012 – 2015
16. Regulation of scale-up of solid state fermentation process for polyunsaturated fatty acids production by fungi. National fellowship program – SAIA, Thailand, 2013
17. New potential of primary products of national plant production, Departmental Project of Ministry of Agriculture, 2013 – 2015
18. Univerzitný vedecký park STU Bratislava, Projekt Štrukturálne fondy EU, Operačný program Výskum a Vývoj, ITMS projektu:26240220084, 2013-2015

Significant papers for last years (totally 112, chapters 8, conference contributions 269 – Invitel lectures - 14)

1. Čertík, M., Andráši, P., Šajbidor, J.: Effect of extraction methods on lipid yield and fatty acid composition of lipid classes containing g-linolenic acid extracted from fungi. J. Am. Oil Chem. Soc., 73(3), 1996, 357-365
2. Čertík, M., Nakahara, T., Kamisaka, Y.: Oleate 6-hydroxylase activity in the membrane fraction from an oleaginous fungus Mortierella ramanniana var. angulispora. Biochem. Biophys. Acta, 1304(1), 1996, 56-64
3. Čertík, M., Baltészová, L., Šajbidor, J.: Lipid formation and g-linolenic acid production by Mucorales fungi grown on sunflower oil. Lett. Appl. Microbiol., 25, 1997, 101-105
4. Pillai, M.G., Čertík, M., Nakahara, T., Kamisaka, Y.: Characterization of triacylglycerol biosynthesis in subcellular fractions of an oleaginous fungus Mortierella ramanniana var. angulispora. Biochem. Biophys. Acta, 1393(1), 1998, 128-136
5. Čertík, M., Sakuradani, E., Shimizu, S.: Desaturase-defective fungal mutants: useful tools for the regulation and overproduction of polyunsaturated fatty acids. Trends Biotechnol., 16, 1998, 500-505
6. Čertík, M., Shimizu, S.: Biosynthesis and regulation of microbial polyunsaturated fatty acid production. J. Biosci. Bioeng., 87(1), 1999, 1-14
7. Čertík, M., Horenitzký, R.: Supercritical CO₂ extraction of fungal oil containing g-linolenic acid. Biotechnol. Techniques, 13(1), 1999, 11-15
8. Čertík, M., Sakuradani, E., Kobayashi, M., Shimizu, S.: Characterization of the second form of NADH-cytochrome b₅ reductase gene from arachidonic acid-producing fungus Mortierella alpina 1S-4. J. Biosci. Bioeng., 88(6), 1999, 667-671
9. Čertík, M., Megová, J., Horenitzký, R.: Effect of nitrogen sources on the activities of lipogenic enzymes in oleaginous fungus Cunninghamella echinulata. J. Gen. Appl. Microbiol., 45(6), 1999, 289-293
10. Čertík, M., Shimizu, S.: Kinetic analysis of oil biosynthesis by arachidonic acid-producing fungus, Mortierella alpina 1S-4. Appl. Microbiol. Biotechnol., 54(2), 2000, 224-230
11. Rapta, P., Polovka, M., Zalibera, M., Breierová, E., Žitňanová, I., Márová, I., Čertík, M.: Scavenging and antioxidant properties of compounds synthesized by carotenogenic yeasts stressed by heavy metals – EPR spin trapping study. Biophysical Chemistry, 116(1), 2005, 1-9
12. Čertík, M., Sláviková, L., Masrnová, S., Šajbidor, J.: Enhancement of nutritional value of cereals with g-linolenic acid by fungal solid state fermentations. Food Technology and Biotechnology, 44(1), 2006, 75-82
13. Breierová, E., Gregor, T., Marová, I., Čertík, M., Kogan, G.: Enhanced antioxidant formula based on a selenium-supplemented carotenoid producing yeast biomass. Chemistry & Biodiversity, 5(3), 2008, 440-446
14. Čertík, M., Adamechová, Z., Hanusová, V., Breierová, E.: Biotechnology as a useful tool for nutritional improvement of cereal-based materials enriched with polyunsaturated fatty acids and pigments. Acta Agronomica Hungarica, 56(4), 2008, 377-384
15. Fakas, S., Čertik, M., Papanikolaou, S., Aggelis, G., Komaitis, M., Galiotou-Panayotou, M.: γ-linolenic acid production by Cunninghamella echinulata growing on complex organic nitrogen sources. Bioresource Technology, 99(13), 2008, 5986-5990
16. Čertík, M., Adamechová, Z.: Cereal-based bioproducts containing polyunsaturated fatty acids. Lipid Technology, 21(11-12), 2009, 250-253
17. Laoteng, K., Čertík, M., Cheevadhanark, S.: Mechanisms controlling lipid accumulation and polyunsaturated fatty acid synthesis in oleaginous fungi. Chemical Papers, 65(2), 2011, 97-103
18. Laho, T., Váradyová, Z., Mihaliková, K., Kišidayová, S., Adamechová, Z., Čertík, M., Jalč, D.: Effects of pre-fermented cereals (ground barley and rye bran) enriched with fungal γ-linolenic acid on rumen fermentation parameters and lipid metabolism in vitro., Journal of Applied Microbiology, 111(3), 2011, 537-546
19. Marova, I., Carnecka, M., Halienova, A., Certik, M., Dvorakova, T., Haronikova, A.: Use of several waste substrates for carotenoid-rich yeast biomass production. Journal of Environmental Management, 95, 2012, 338-342
20. Salar, R.K., Certik, M., Brezova, V.: Modulation of phenolic content and antioxidant activity of maize by solid state fermentation with Thamnidium elegans CCF 1456. Biotechnology and Bioprocess Engineering, 17(1), 2012, 109-116
21. Zorádova Murínová, S., Dudášová, H., Lukáčová, L., Čertík, M., Šilhárová, K., Vrana, B., Dercová, K.: Adaptation mechanisms of bacteria during the degradation of polychlorinated biphenyls in the presence of natural and synthetic terpenes as potential degradation inducers. Applied Microbiology and Biotechnology, 94(1), 2012, 1375 - 1385
22. Čertík, M., Adamechová, Z., Laoteng. K.: Microbial production of gamma-linolenic acid: Submerged versus solid-state fermentations. Food Science and Biotechnology, 21(4), 2012, 921-926
23. Čertík, M., Breierová, E., Oláhová, M., Šajbidor, J., Márová, I.: Effect of selenium on lipid alternations in pigment-forming yeasts. Food Science and Biotechnology, 22(S), 2013, 45-51
24. Mike, N., Papp, G., Certik, M., Czibulya, Z., Kunsági-Máté, S.,  Ember, I., Vágvölgyi, C., Pesti, M., Gazdag, Z.: Regulation of cytotoxic, non-estrogenic, oxidative stress-induced processes of zearalenone in the fission yeast Schizosaccharomyces pombe. Toxicon, 73, 2013, 130-143
25. Čertík, M., Adamechová, Z., Guothová, L.: Simultaneous enrichment of cereals with polyunsaturated fatty acids and pigments by fungal solid state fermentations. Journal of Biotechnology, 168, 2013, 130-134
26. Klempova T., Mihalik, D., Certik, M.: Characterization of membrane bound fatty acid desaturases. General Physiology and Biophysics, 32, 2013, 445-458
27. Klempova, T., Basil, E.,, Kubatova,  A., Certik, M.: Biosynthesis of gamma-linolenic acid and beta-carotene by Zygomycetes fungi. Biotechnology Journal, 8, 2013, 794-800
28. Čertík, M., Klempová, T., Guothová, L., Mihálik, D., Kraic, J.: Biotechnology for the functional improvement of cereal-based materials enriched with polyunsaturated fatty acids and pigments. European Journal of Lipid Science and Technology, 115(11), 2013, 1247-1256
29. Cibulková, Z., Čertík, M., Dubaj, T.: Thermooxidative stability of poppy seeds studied by non-isothermal DSC measurements. Food Chemistry, 150, 2014, 296-300
30. Mihalik, D., Gubisova, M., Klempova,T., Certik, M., Ondreickova, O., Hudcovicova, M., Klcova, L., Gubis, J., Dokupilova, I., Ohnoutkova L., Kraic, J.: Transformation of barley with artificial gene encoding for delta-6-desaturase. Biologia Plantarum, 2014, accepted

Book chapters:

1. Čertík, M., Shimizu, S.: Progress in polyunsaturated fatty acid production by fungi. In: Recent Research Developments in Oil Chemistry (Pandalai, S.G., ed), Transworld Research Network, Trivandrum, India, Vol. 2, 1998, 89-103. (ISBN: 81-86846-15-8) (CA: 130-278990g)
2. Čertík, M.: Fermentation physiology and regulation of microbial polyunsaturated fatty acid biosynthesis. In: Research Advances in Bioscience & Bioengineering (Mohan, R.M. ed.), Global Research Network, Kerala, India, Vol. 1, 2000, 45-64. (ISBN: 81-87736-16-X)
3. Čertík, M.: Biotransformation of oils to value-added compounds. In: Biocatalysis and Bioenergy (Hou, C.T., Shaw, J.-F., eds.,). John Wiley & Sons, New Jersey, USA, 2008, 571-585. (ISBN: 978-0-470-13404-7)
4. Čertík, M., Hanusová, V., Breierová, E., Márová, I., Rapta, P.: Biotechnological production and properties of carotenoid pigments. In: Biocatalysis and Agricultural Biotechnology (Hou, C.T., Shaw, J.-F., eds.,). Tailor and Francis-CRC Press, Boca Raton, FL, USA, 2009, 355-375 (ISBN: 1420077031, 9781420077032)
5. Laoteng, K., Čertík, M.: Biotechnological production and application of high-value microbial oils. In: Industrial Fermentation: Food Processes, Nutrient Sources and Production Strategies (Krause, J., Fleischer, O., eds.). Nova Science Publisher, Inc., Hauppauge (NY, USA), 2010, 187-215 (ISBN: 978-1-60876-550-8)
6. Čertík, M., Adamechová, Z., Sláviková, L.: Biotechnological enrichment of cereals with polyunsaturated fatty acids. In: Biocatalysis and Biomolecular Engineering (Hou, C.T., Shaw, J.-F., eds.,). John Wiley & Sons, Hoboken (NJ, USA), 2010, 175-193 (ISBN: 978-0-470-48759-4)
7. Márová, I., Čertík, M., Breierová, E.: Production of Enriched Biomass by Carotenogenic Yeasts – Application of Whole-Cell Yeast Biomass to Production of Pigments And Other Lipid Compounds. In: Biomass - Detection, Production and Usage. Darko Matovic (Ed.), InTech, 2011, 345-384 (ISBN: 978-953-307-492-4)
8. Obruča, S., Márová, I., Čertík, M.: Whey: Attractive Substrate for Microbial Production of Substances and Materials for Medical Applications. In: Whey: Types, Composition and Health Implications (Rafael Mauro Benitez, Gustavo M. Ortero, Eds.) Nova Publishers (New York, USA), 2012, 53-92 (ISBN: 978-1-61942-862-1)

 

Collaborations:

Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia

Plant Production Research Center, Piešťany, Slovakia

Institute of Animal Biochemistry and Genetics, Slovak Academy of Sciences, Ivanka pri Dunaji, Slovakia

Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia

Universtity of Veterinary Medicine and Pharmacy, Košice, Slovakia

Faculty of Chemistry, Brno University of Technology, Czech Republic

King Mongkut's University of Technology Thonburi (KMUTT), Bangkok, Thailand

National Center for Genetic Engineering and Biotechnology (BIOTEC), Bangkok, Thailand

Faculty of Sciences, University of Pécs, Pécs, Hungary

Centre National de la Recherche Scientifique (CNRS), Micalis, Jouy-en-Josas, France

Institute of Biochemistry, Technische Universität Graz, Graz, Austria

University of Patras, Patras, Greece

 

Photos:

Gene engineering laboratory

Analytical laboratory (GC/MS/FID, HPLC/DAD/RI, TLC/UV-VIS scanner, UV-VIS spectrofotometer)

Laboratory for cultivation of microorganisms and extraction/purification of lipophilic compounds