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DEPARTMENT OF MICROBIAL BIOSYNTHESIS AND BIOTECHNOLOGIES
Mladenka Paunova Ilieva-Stoilova, DSc, ProfessorHead of Department
Tel: (+359 32) 642 430E-mail: [email protected]
1939 Born March 24, Kostinbroad, Bulgaria1962 MS, Faculty of Chemistry, Sofia University ‘St. Kliment Ohridski’1963 Research Scientist, Higher Institute of Food and Flavour Industries, Plovdiv1973 PhD in Biotechnology, Higher Institute of Food and Flavour Industries1980 Associate Professor, Higher Institute of Food and Flavour Industries1982 Associate Professor, Institute of Microbiology, BAS2000 DSc in Biotechnology, Institute of Microbiology, BAS1993 Coordinator of the Research Group in Plovdiv, Institute of Microbiology, BAS2000 DSc, Institute of Microbiology, BAS2005 Head of the Department of Microbial Biosynthesis and Biotechnologies, Institute
of Microbiology, BASTeaching: 1963-95 Higher Institute of Food and Flavour Industries
STAFF
Bonka Tzvetkova, PhD, Associate Professor [email protected] Alexander Ratkov, PhD, Associate ProfessorPenka Alexieva, PhD, Associate Professor [email protected] Veneta Ivanova, PhD, Associate Professor [email protected] Emilina Simova, PhD, Associate Professor [email protected] Christozova, PhD, Associate Professor [email protected]; lbpmbas @ yahoo . com Dora Beshkova, PhD, Associate Professor [email protected]; [email protected] Kostantza Pavlova, PhD, Associate Professor [email protected]; lbpmbas @ yahoo . com Ginka Frengova, PhD, Associate Professor lbpmbas @ yahoo . com Adriana Gousterova, PhD, Associate Professor marine _ bg @ abv . bg Atanas Pavlov, PhD, Associate Professor [email protected]; [email protected] Victoria Gesheva, PhD, Research Scientist [email protected] Vessela Raykovska, MS, Research ScientistLiliana Nacheva, MS, Research Scientist lin 1@ abv . bg Dimka Tuneva, PhD, Research Assistant lbpmbas @ yahoo . com Mariana Kolarova, MS, Research AssistantKrasya Aleksieva, MS, Research AssistantVelitchka Ivanova, MS, Research AssistantJulia Krusteva, MS, Research AssistantPhilip Philipov, MS, Research AssistantDonka Paskaleva, MS, Research AssistantKalin Todorov, MS, Research AssistantMart Mihailov, MS, Research AssistantIgnat Dimov, MS, Research AssistantTatyana Gocheva, MS, Research Assistant lbpmbas @ yahoo . com Vasil Georgiev, MS, Research Assistant [email protected]; [email protected] Svetla Kostadinova, Research Assistant [email protected] Elka Genova, Laboratory Assistant [email protected] Stefka Vassileva, Laboratory AssistantVasilkaUsherova, Laboratory Assistant [email protected]
MAIN RESEARCH TOPICS
The research is concentrated on the metabolism of prokaryotic and eukaryotic cells and the possibilities for its regulation. On this base the biotechnology of production of biologically active substances for medicine and starters and additives for food industry are developed. The following aspects are studied:
Yeasts – physiology and biochemistry of methylotrophic, beaker’s and fodder’s yeast and biosynthesis of biologically active substances; biological diversity of psyhrophilic yeasts from Livingston island – Antarctica and production of biologically active substances (enzymes, lipids and polysaccharides) by them.
Actinomycetes and filamentous fungi - physiology, biochemistry, biosynthesis and identification of primary and secondary metabolites. The investigations are focused on: Actinomycetes – taxonomy, physiology, biochemistry and biosynthesis of enzymes, antibiotics and stimulators; Isolation, purification and structure of biologically active substances; Filamentous fungi – optimization of conditions for production of extracellular hydrolytic enzymes by free and immobilized cells.
Characteristics of the obtained enzyme preparations and investigation of possibilities of their application.
Lactic acid bacteria - physiology and resources for development of dairy products whit improved nutritional value. The work is focused on the physiology and resources of lactic acid bacteria, and health solutions. That involves creation of effective bio-consortium cultures for fermented milk with improved physiological and nutritional value by: Incorporation of antimicrobial agents with wide inhibitory spectrum to selected starter cultures for implementation of the protective functions and of biocontroling agents – regulation of the microflora, directing of the processes of forming and ripening of Bulgarian dairy products and participation in the biotransformation of the milk substances; The proteolytic systems of lactic acid bacteria and association of selected lactobacillus strains with high peptidolytic profilesis studied. New approaches for development of dairy products with high biological (health) benefits and nutritive value, quality and safety.
Amino acids – biotechnologies for their production.
Biological active substances by plant in vitro systems. The possibilities of different plant in vitro systems for the production of low molecular weight biologically active substances (including additives and antioxidants for food industries), enzymes and polysaccharides are investigated. The research concerns optimization of the biosynthetic processes, isolation, purification and identification of the obtained metabolites. Special attention is drawn to application of different strategies for enhancement of the yield and effectiveness of biotechnological processes (selection, elicitation, two-phase cultivation etc.).
MOST IMPORTANT ACHIEVEMENTS
A method for production of extra cellular α-galactosidase was developed by submerged cultivation of Aspergillus niger M47 , Penicillium chrisogenum LN33 and Humicula lutea 1205 on the medium, composed by wastes from food industry. The method was submitted for patent.
A Lab biotechnology for galanthamine production by Leucojum aestivum shoot culture was developed. This is the first bioreactor process reported for production of Galanthamine by in vitro plant system.
A detailed “know-how” for production of L-lysine and a Lab biotechnology for production of L-valine were developed.
From culture medium of a thermophilic strain two indole alkaloids (microbiareton and microbiaeratinin) were isolated and their cytotoxic and antialgic activities were established.
An alkaline hydrolysate from animal wastes was obtained and their influence on chemical and microbiological properties of the poor soils were investigated. The obtained results outlined the possibilities for its application as an organic fertilizer.
The growth of methylotrophic yeasts Hansenula and Pichia on glucose, methanol and glycerol as carbon source was investigated. The presence of catalase and superoxide dismutase (SOD) activity in fractions of mitochondria, cytosol and peroxisomes were investigated. It was established that peroxisomal catalase was located in the mitochondrial matrix and SOD( besides the typical places for localization- mitochondria and cytosol)was detected in peroxisomes as well.
The proteolytic system of starter lactobacteria was characterized and it was established that this system is a criterion for their participation in starters for milk products.
In situ baceriocins production of LB. bulgaricus BB18 and LB. lactis BCM5 was studied and their strain compatibility with two technological strains S. thermophilus 11A and Lb. bulgaricus LX1 was demonstrated for forming starters for fermented milks with health benefits.
The strain Streptomyces sp., producer of antibiotic complex, was identified by hemotaxonomic and genetic methods as Streptomyces iakurus DSM 41873. The antibiotic complex is composed of 5 new components, which were characterized by their molecular masses and chemical structures.
The colloid properties of glucomannan, synthesized by psychrophilic stain were studied. The comparison of the cream emulsions with glucomannan and those with emulsifiers rofetan and arlacel (used in cosmetic industry) showed that they were with equal stability.
The yeast strain, producer of phytase was isolated from Antarctic samples. The appropriate culture medium and parameters of cultivation were established for high production of phytase. The enzyme kinetics and properties were determined as well.
By flow cytometry the genom sizes of the cells of Beta vulgaris intact plants and hairy root cultures were determined. It was established that in contrast to the primary plant tissues, the obtained hairy roots are mixoploids., due to several cycles of endoreduplication. Such difference currently is not reported in the available scientific literature.
SELECTED PUBLICATIONS
Ilieva, M., Pavlov, A., Kovatcheva, E. (2002): Further Research into Lavandula Species: Cell Cultures of Lavandula vera and Rosmarinic Acid Production - In: Genus Lavandula – series “Medicinal and Aromatic Plants (M. Lis-Balchin - ed) – Taylor & Francis Publishers, 214-226– ISBN 0-415-28486.
Hristozova, Ts., Michailova, L., Tuneva, D., Gotcheva, V., Angelov, A., Roshkova, Z. (2002): Mutant Hansenula polymorpha with constitutive alcohol oxidase and peroxisome biosynthesis. Z. Naturforsch. 57c, 858-862.
Hristozova, Ts., Rasheva, T., Nedeva, T., Kujumdzieva, A. (2002): Superoxide dismutase duringng glucose repression of Hansenula polymorpha CBS 4732. Z. Naturforsch. 57c, 313-318.
Hoshino,Y., Ivanova, V., Yazawa, K., Ando, A., Mikami, Y., Zaki, S., Karam, Z., Youssef, Y., Graefe, U. (2002): Queenslandon, a new antifungal compound produced by Chrysosporium queenslandicum: Production, isolation and structure elucidation. J. Antibiotics 55, 516-520.
Ivanova,V., Aleksieva, K., Kolarova, M., Chipeva, V., Schlegel, R., Schlegel, B., Graefe, U. (2002): Neuropogonines A, B and C, new depsidon-type metabolites from Neuropogon sp., an Antarctic lichen. Die Pharmazie 57, 73-74.
Ivanova, V., Hoshino, Y., Yazawa, K., Ando, A., Mikami, Y., Graefe, U. (2002): Isolation and structure elucidation of two new antibacterial compounds, produced by Chrysosporium queenslandicum. J. Antibiotics 55, 914-919.
Simova, E., Beshkova, D., Angelov, A., Hristozova, T., Frengova, G., Spasov, Z. (2002): Lactic acid bacteria and yeast in kefir grains and in kefir obtain with them. J. Ind. Microbiol. Biotechnol. 28, 1-6.
Gesheva, V. (2002): Rhizosphere microflora of some citrus as a source of antagonistic actinomycetes. Eur. J. Soil Biol. 38, 85-88.
Aleksieva, P., Spasova, D., Radoevska, S. (2003): Acid phosphatase distribution and localization in the fungus Humicola lutea. Z. Naturforsch. 58c, 239-243.
Aleksieva, P., Tchorbanov, B., Michailova, L., Nacheva, L. (2003): Improvement of acid phosphatase production by immobilization of Humicola lutea mycelium in polyurethane sponge. World J. Microbiol. Biotechnol. 19, 247-253.
Berkov, S., Pavlov, A. (2004): A rapid densitometric method for the analysis of hyoscyamine and scopolamine in solanaceous plants and their transformed root cultures. Phytochem Analysis 15, 141-145.
Simova, E., Frengova, G., Beshkova, D. (2004): Exopolysaccharides produced by mixed culture of yeast Rhodotorula rubra GED10 and yogurt starter (Strepto-coccus thermopilus 13a + Lactobacillus bulgaricus 2-11). J. Appl. Microbiol. 97, 512-519.
Pavlova, К., Koleva, L., Kratchanova, M., Panchev, I. (2004): Production and charactereization of an exopolysaccharide by yeast. World J Microbiol. Biotechnol. 20(4), 435-440.
Goshev, I., Gousterova, A., Vasileva-Tonkova, E., Nedkov, P. (2005): Characterization of the enzyme complexes produced by two newly isolated thermophylic actinomycete strains during growth on collagen-rich materials. Process Biochem. 40, 1627-1631.
Gousterova, A., Braikova, D., Goshev, I., Christov, P., Tishinov, K., Vasileva-Tonkova, E., Haertlé, Т., Nedkov, P. (2005): Degradation of keratin and collagen containing wastes by newly isolated thermoactinomycetes or by alkaline hydrolysis . Lett. Appl. Microbiol. 40, 335-34.
Angelov, A., Gotcheva, V., Hristozova, Ts., Gargova, S. (2005): Application of pure and mixed probiotic lactic acid bacteria and yeast cultures for oat fermentation. J. Science Food and Agriculture 85, 2134-2141.
Gesheva, V. (2005): Microbiota of subantarctic soils from South Georgia Island. Cyprus J. Sci. 3, 75-81.
Gesheva, V., Ivanova, V., Gesheva, R. (2005): Effects of nutrients on the production of AK-111-81 macrolide antibiotic by Streptomyces hygroscopicus. Microbiological Research 160, 243-248.
Pavlov, A., Popov, S., Kovacheva, E., Georgiev, M., Ilieva, M. (2005): Volatile and polar compounds in Rosa damascene Mill 1803 cell suspension. Journal of Biotechnology 118, 89-97.
Pavlov, A, Georgiev, V, Ilieva, M. (2005): Betalain biosynthesis by red beet (Beta vulgaris L.) hairy root culture. Process Biochemistry 40, 1531-1533.
Simova, E., Simov, Zh., Beshkova, D., Frengova, G., Dimitrov, Zh., Spasov, Z. (2006): Amino acid profiles of lactic acid bacteria, isolated from kefir grains, and kefir starter made from them. Int. J. Food Microbiol. 107, 112-123.
Simova, E., Beshkova, D. (2006): Effect of growth phase and growth medium on the peptidase activities of starter lactobacteria. Le Lait, Dairy Science Technol. (in press).
Hristozova, Ts., Angelov, A., Tzvetkova, B., Paskaleva, D., Gotcheva, V., Gargova, S., Pavlova, K. (2006): Effect of furfural on carbon metabolism key enzymes of lactose-assimilating yeasts. Enzyme Microb. Technol. 39,1108-1112.
Georgiev, M., Kuzeva, S, Pavlov A., Kovacheva, E., Ilieva, M. (2007): Elicitation of rosmarinic acid by Lavandula vera MM cell suspension culture with abiotic elicitors - Word Journal of Microbiology and Biotechnology 23, 301-304.
Pavlov, A., Berkov, S., Courot, E., Gotcheva, T., Tuneva, D., Pandova, B., Georgiev, M., Georgiev, V., Yanev, S., Burrus, M., Ilieva, M. (2007): Galanthamine production by Leucojum aestivum in vitro systems, Process Biochem. in press, doi: 10.1016/j.procbio.2006.12.2006
Georgiev, M.., Pavlov, A., Bley, T. (2007): Hairy root type in vitro systems as sources of bioactive substances. Appl. Microbiol. Biotechnol. (in press), DO1 1o.1007/300253-007-0856-5.