Anna T. Salek, Ph.D., D.Sc. (Dr. habil.)
TOPICS OF EXPERTISE AND RESEARCH
I. Hands-on experience in food biotechnology, i.e.:
- in microbial physiology (e.g. optimalisation of growing and fermentation processes),
- in biochemistry of enzymes and proteins (i.e., development, recovery and purification
- of protein: analytical and preparative scale), in regulation of microbial metabolism,
- in molecular cell biology,
- in applied microbiology, especially on the improvement of bacteria and yeast strains, both in the laboratory and in industry scale (Probiotics in Functional Food),
- in Food Safety Microbiology (HACCP-System for pathogens, Risk Assessment, Microbial Food Monitoring, International Food Standard).
- In environment (water and air): new methods of identification (biosensors).
- Work on the bacteria Escherichia coli, Enterobacter aerogenes and Salmonella typhimurium isolated from dairy products, i.e., microbiological and biochemical diagnostic, a chemical and a physical mutagenesis of cells, regulation of bacterial metabolism, especially work on biochemistry of ß-galactosidase and pyruvate ehydrogenase complex. Results were applied in cheese-making (see papers in “Publication List”).
- Microbiological work on the technological aspects of dairy products, optimalisation of fermentation processes (beer & alcohol beverage, mathematical modelling), probiotics and functional food as well as work on the animal feeds (probiotics, starter culture) (see 4 patents and papers in “Publication List”).
- Genetic work on the killer protein of yeast – secretion system (especially on bakery and wine strains), constructed by electro-transformation of dsRNA (and also co-electro-transformation with plasmid pADH) or of „Virus Like Particles“ with dsRNA into cells (stipend of UNDP/FAO- Roma and project at the University of Würzburg, Institute of Microbiology and Genetics, Department of Biotechnology, Germany). See papers in “Publication List”. Strains developed in ways described above were used in the food technology, in the agriculture, in the industrial fermentation processes and in the production of baker’s yeast as well as for alcoholic beverages, during the course of collaborative work (consulting and application of new strains) with the fermentation industry in Poland (the former government-owned „Polmos“ companies and the Institute of Biotechnology in Warsaw, Poland) from 1980 up to now.
- Work on the cytoskeleton of yeast cells (cell biology): a new technique that was developed and used within my research was enucleation, i.e., technique for the removal of nucleus from cell/protoplasts of Saccharomyces cerevisiae (including fluorescence microscopic analyses, micrography). It was used for the construction of cybrids (by electro-fusion technique), i.e., new yeast hybrids possessing mixed cytoplasms but only one nucleus after enucleation; such cybrids could be designed to obtain new yeast industry strains (for food biotechnology) with particular abilities, such as that of secretion of certain molecules (and also for research on RNA or DNA-viruses, i.e., VLPs). Work at the University of Würzburg, Institute of Genetic and Microbiology, Germany (see papers in “Publication List”).
- Biochemical work on the purification of killer yeast protein (glycoproteins), i.e., gel filtration, ion exchange chromatography, isoelectrofocusing (analytical and preparative-scale), as well as PAGE (native or with SDS, IPG/gradient pH or Gradient PA), 2-D(2-dimensional electrophoresis, i.e. IPG/2-D-SDS PAGE), an ultrafiltration and concentration („Amicon“ systems and freeze-drying/lyophilisation techniques) of proteins and also different methods for dialysis process, etc. Purified killer Protein could be used as biological control for pathogen infections. Work at the Technical University of Munich-Weihenstephan, Germany (see papers in “Publication List”);
- Work on the PCR (molecular biology) (Inter-LINE-PCR as well as on the RAPD-PCR and AP-PCR) and on the separation of chromosomes (by PFGE, pulse field gel electrophoresis) for the characterisation of medical and industrial: brewing, wine and bakery strains (at the GSF – National Research Centrum for Environment and Health, GmbH in Munich, and at the Technical University of Munich-Weihenstephan, Institute of Technical Microbiology and Technology of Brewing, Germany, respectively). Results were used for development of automated yeast identification system by the application of PCR technique (see papers in “Publication List”). Most of my research in cell biology has been on Saccharomyces spp., but some was on Kluyveromyces spp. and especially recently – on Pichia (Hansenula) spp. (with expression of killer protein-toxin, source of dsRNA and dsDNA plasmids), Candida spp. and Fusarium spp., which exist also in food (especially in wine).
- Food & Environment Safety Microbiology and Health Protection System: Food Safety, HACCP, IFS, GMO, Functional Food, Nutraceuticals. Routine & research work on biochemical, microbiological and immunological identification of pathogenic microorganisms in drink and waste water (bacteria, yeasts and fungi, i.e. Cryptosporidium parvum, Campylobacter, Escherichia coliserotype O157:H7 (EHEC), Legionella pneumophila, Candida albicans, Aspergillus fumigatus, Fusariums, etc.) as well as in food products (Salmonella, Listeria, Escherichia coli serotype O157:H7 (EHEC) and Campylobacter jejuni). Work for analytical laboratory of food products and environment, using Food Safety and Protection System, i.e. HACCP (Hazard Analysis Critical Control Point and predictive microbiology), microbiological monitoring and modern, rapid & safety methods (e.g. immunology, genetic, bio-sensors). For this case see paper (Salek A.:
A modern microbiology methods in food monitoring, HACCP systems and in diagnostics. The Journal of Food Industry. 2004).
II. Experience in teaching of food biotechnology
I have had several years experience in the teaching of general microbiology, food biotechnology (aspects of molecular biology), as well as food technology (functional food, especially dairy and yeast fermentation) at the University of Warsaw and also in industry (see enclosed CV) as well as at the University Würzburg and Munich (Germany).
I had also many congresses, seminars, written monographs and summer-schools for students and industrial microbiologists on the microbiological topics, i.e. applications of molecular biology in food technology, about transgenic and new microorganisms, like Legionella pneumophila, Cryptosporidium parvum or about biological control of pathogen infections or biofilm in practice (water in different systems).
III. Hands-on experience in biotechnology of pharmaceutical, i.e.:
- in molecular cell biology of yeast and bacteria,
- in biochemistry of yeast proteins (i.e. development, recovery and purification of recombinant proteins: analytical and preparative scale), in regulation of yeast metabolism,
- in yeast and bacteria physiology (optimalisation of growing and Fermentation processes),
- in applied of killer-yeast-proteins (drug-glycoprotein), especially in the industry scale,
- vet. medicine safety – microbiological control of pharmaceutical products, according to Ph. Eur. 2001.
- Regulatory Affairs (MRLs) work on the pharmacology (pharmacodynamics and pharmacokinetics: absorption, distribution, metabolism, excretion) as well as toxicology (toxicity, mutagenicity, teratogenicity, carcinogenicity) of yeast killerprotein using mammalian cells culture (killer toxin as a new antimicrobial agents for oral application, against pathogenic bacteria, yeasts and fungi, infected domestic animals); work at the firm „Inropharm“ GmbH & Co. and Technical University of Munich, Germany (see papers in “Publications List”).
- Registration of new antimicrobial agent and veterinary medicine in EU (Report on the Safety and Residue File) for EU in Brussels/London (EMEA), work as Regulatory Affair Scientist for „Inropharm“ GmbH & Co., Germany (3 Monographs) and for German Ministry of Health in Berlin (2 Monographs).
- Work at the pharmaceutical firm for a production and validation of methods for chemical analysis of veterinary medicines („Inropharm“ GmbH & Co., Germany).
- Sterility (Sterilität) activity of preservative (Konservierungsstoffe) control of veterinary products (especially medicines for injection), according to book Pharm. Eur. 2001, 2002.
- General hygiene control in pharmacy industry, according to GMP/GHP requirements.
IV. Experience as manager of food biotechnology and as pharmaceutical and environmental laboratory
Over 10 years I was the head of microbiology laboratory at the Agricultural University of Warsaw as well as at the Institute of Biotechnology (Warsaw, Poland) and while there, I managed several research projects in biotechnology area. Next, over 28 years I was the head of microbiology laboratory: at the pharmaceutical firm, environmental firm and also at the Microbiology Institute (Germany) as head of Food & Environment & Veterinary & Feed Microbiology, Biotechnology (incl. veterinary with Bovine Spongiforme Encephalopathy, BSE) and Immunology Laboratory. Last 12 years I have been manager Microbiology & Chemistry Labor for Analytic of Water & Environment (incl. fungi diagnostic). It was work on Legionella pneumophila problematic (Routine and Research with patent About new method for rapid identification biofim-bacteria, incl. Legionella pneumophila).
V. Future research in food & environment & pharmaceutical biotechnology
As a microbiologist with hands-on experience in the application of novel techniques to cell engineering, I have worked (patent) on the use of naturally-produced peptides or proteins from yeasts for the therapeutic treatment of infected animals, i.e. for diseases caused by pathogenic bacteria, yeasts and fungi (use of killer yeast toxins for biological control).
I have established strong evidence that some yeast toxins are good anti-microbial agents (like probiotics) and can be effective, as pre-therapeutics in the treatment of antibiotic-resistant intestinal (E.coli, Enterobacter aerogenes, Salmonella typhimurium, Enterococcusspp., etc.) and other infections (e.g. Styphylococcus aureus, Pasteurella haemolytica, Candida albicans, Acremonium or Aspergillus fumigatus, etc.) in several domestic animals (see papers in “Publications List”).
It could be a big interest to continue farther this topic. Future research in biotechnology they are modern & rapid & safety instrumental as well as electrochemical methods, connected with experiences of follow sciences:
- molecular biology
- biophysics and
See paper (Salek A.: A modern microbiology methods in food monitoring, HACCP systems and in diagnostics. The Journal of Food Industry. 2004).
VI. Experience in accreditation of biological laboratory according to ISO 17025:2008 norm
In 1999, 2000, 2004, 2005, 2010 und in 2014 were prepared documents (QMH, SOP´s and others) for accreditation microbiological and chemical laboratory (in water and food technology
field) according to ISO norm 17025:2005. Accreditation Office: DAP (now DAkkS), Berlin, Germany.