Ilya Tolstorukov, PhDResearch Professor
Areas of Expertise
Pichia pastoris, P.methanolica, Hansenula polymorpha, S.cerevisiae, Yeast Molecular Biology, Microbial Genetics and Biotechnology, Regulation of Gene Expression, Control of Mating Type in Yeasts, Yeast Lipid Metabolism, Genomics, Recombination, Expression of Recombinant Proteins, Production of Antibodies in *P.pastoris*, Isothermal DNA Amplification, Biology and Genetics of Methylotrophic Yeasts
Dr. Tolstorukov earned his degrees in genetics, PhD in 1973 and DSc in 1989, in the Soviet Biotechnological Industry's State Institute of Genetics and Selection of Industrial Microorganisms ("NIIGenetika") in Moscow, USSR. In 1992 by decision of the Higher Attestation Commission of the USSR he was awarded the title of full Professor in Genetics. Before coming to Keck Graduate Institute of Applied Life Sciences in 2001, Dr. Tolstorukov spent more than 30 years working as Researcher, Group Leader, Professor and Section Head at the NIIGenetika. He was invited as Visiting Scientists into National Cancer Research Institute, Frederick, MD, USA (1989-1990) and University of Groningen, Haren, the Netherlands (1995), and as Visiting Professor into Osaka University, Osaka, Japan (1997-1998). During 1976-1998 he consulted several Soviet state plants manufacturing yeast as a source of single cell protein for animal feed and a number of privet biotechnological businesses in Russia.
His discovery of a complex genetic system controlling homo- and heterothallism in yeast (PhD thesis, 1969-1974) has had a substantial impact on the development of the "cassette model" of mating type interconversion in S. cerevisiae and the discovery of gene silencing in yeast. Beginning from 1975, Dr. Tolstorukov pioneered the genetic investigations of methylotrophic yeast species. His work with Pichia methanolica resulted in the discovery of genetic manipulation methods of the yeast sexual cycle and the development of other classical genetic methods for this group of yeasts, including mutagenesis, mating, complementation, tetrad analysis and plasmid transformation (Tolstorukov et al., 1977-1994). Dr. Tolstorukov's work served researchers in Western Europe and the United States, including Dr. James Cregg, as a template for developing methods for genetic manipulations for other methylotrophic yeast species, Hansenula polymorpha and Pichia pastoris. Dr. Tolstorukov holds several patents issued in the Soviet Union.
In 2001 Dr. Tolstorukov was invited as a Visiting Research Scientist to KGI. He became a Senior Research Scientist and later was appointed as Research Professor at the Dr. Cregg's lab and has taken responsibilities for most of the academic and applied projects in the lab related to the expression of proteins in the methylotrophic yeast Pichia pastoris and the improvement of the Pichia pastoris expression system. Dr. Tolstorukov developed a novel proprietary technology for manufacturing antibodies in yeast (US patents issued in 2011-2012). He also initiated projects for analysis of new promoters for expression of heterologous proteins in Pichia pastoris. He has cloned and demonstrated potential efficiency of a number of such promoters (US Patent issued in 2012 and patent applications filed in 2012). As co-founder and principal scientist of Biogrammatics, Inc., a start-up company working on improvement of the Pichia pastoris expression system, Dr. Tolstorukov has been involved in construction of a novel user-friendly host-vector system for this yeast as well as in creating Pichia pastoris producers expressing proteins of interest for a number of academic laboratories and companies.He currently works on several research projects related to the application of the P. pastoris expression system for production of important proteins, such as phytase - an animal feed enzyme widely used in the diets of pigs and chickens, alpha- and beta-galactosidases, that have potentials in using as animal and human food additives. He is also working in collaboration with the lab of Dr. Angelika Niemz to construct, clone and sequence of targeting DNA molecules, grow microorganisms of interest and establish the PROXAR reaction (Isothermal PROXimity DNA Amplification Reaction) for DNA-based detection of multidrug resistance mutations in Mycobacterium tuberculosis.
- Selected Publications
- Naumov GI, Tolstorukov II. "Comparative Genetics of Yeasts .Communication X. Reidentification of Mutators of Mating Types in Saccharomyces". Genetika 1973;9(1):82-91.
- Tolstorukov II, Dutova TA, Benevolensky SV, Soom YO. "Hybridization and Genetic-Analysis of Methanol-Utilizing Yeast Pichia Pinus". Genetika 1977;13(2):322-329
- Tolstorukov II, Benevolensky SV, Efremov BD. "Genetic-Control of Cell Type and Complex Organization of the Mating Type Locus in the Yeast Pichia-Pinus". Current Genetics 1982;5(2):137-142
- Tolstorukov II, Efremov BD. "Genetic-Mapping of the Yeast Pichia-Pinus. Communication II. Mapping by Tetrad Analysis". Genetika 1984;20(7):1099-1107
- Nikoshkov AB, Kistanova EN, Eremaschvili MS, Tolstorukov I. "Expression of the human leukocyte interferon A gene in Saccaromyces cerevisiae yeast under the control of yeast genes PHO5, GALI, and GAL10 regulatory elements". Molecularnaya Genetika, Microbiologia and Virusologiya 1987;6:42-45.
- Sibirny AA, Titorenko VI, Efremov BD, Tolstorukov II. "Multiplicity of Mechanisms of Carbon Catabolite Repression Involved in the Synthesis of Alcohol Oxidase in the Methylotrophic Yeast Pichia-Pinus". Yeast 1987;3(4):233-241
- Kushnirov VV, Teravanesyan MD, Didichenko SA, Smirnov VN, Chernoff Y, Derkach IL, Novikova ON, Ingevechtomov SG, Neistat MA, Tolstorukov II. "Divergence and Conservation of Sup2(Sup35) Gene of Yeasts Pichia-Pinus and Saccharomyces-Cerevisiae". Yeast 1990;6(6):461-472.
- Tarutina MG, Tolstorukov II. "Development of A Method for the Vector Transformation of the Methylotrophic Yeast Pichia-Methanolica". Genetika 1994;30(6):783-790.
- Anamnart S, Tolstorukov I, Kaneko Y, Harashima S. "Fatty acid desaturation in methylotrophic yeast Hansenula polymorpha strain CBS 1976 and unsaturated fatty acid auxotrophic mutants". Journal of Fermentation and Bioengineering 1998;85(5):476-482
- Lu SF, Tolstorukov II, Anamnart S, Kaneko Y, Harashima S. "Cloning, sequencing, and functional analysis of H-OLE1 gene encoding Delta 9-fatty acid desaturase in Hansenula polymorpha". Applied Microbiology and Biotechnology 2000;54(4):499-509.
- Tarutina MG, Tolstorukov II. "Mechanisms of transformation in yeast Pichia methanolica: Transforming and nontransforming genes". Russian Journal of Genetics 2001;37(1):54-65.
- Lahtchev KL, Semenova VD, Tolstorukov II, van der Klei I, Veenhuis M. "Isolation and properties of genetically defined strains of the methylotrophic yeast Hansenula polymorpha CBS4732". Archives of Microbiology 2002;177(2):150-158.
- Lin-Cereghino GP, Godfrey L, de la Cruz BJ, Johnson S, Khuongsathiene S, Tolstorukov I, Yan MD, Lin-Cereghino J, Veenhuis M, Subramani S, et al. "Mxr1p, a key regulator of the methanol utilization pathway and peroxisomal genes in Pichia pastoris". Molecular and Cellular Biology 2006;26(3):883-897.
- Tolstorukov I, Cregg JM. "Classical Genetics". Methods of Molecular Biology 2007;389:189-202.
- Sunga J, Tolstorukov I, Cregg JM. "Post-transformational vector amplification in the yeast Pichia pastoris". FEMS Yeast Research 2008;8(6):870-876.
- Cregg JM, Tolstorukov I, Kusari A, Sunga J, Madden K, Chappell T. E"xpression in the Yeast Pichia Pastoris". Guide to Protein Purification, Second Edition 2009;463:169-189.
- Cregg J, Latham J, Litton M, Schatzman R, Tolstorukov I. (2011). Methods of synthesizing heteromultimeric polypeptides in yeast using a haploid mating strategy. US Patent 7,927,863.
- Cregg J, Tolstorukov I. (2012). P. pastoris ADH promoter and use thereof to direct expression of proteins. US patent 8,222,386.
- Cregg J, Latham J, Litton M, Schatzman R, Tolstorukov I. (2012). Methods of synthesizing heteromultimeric polypeptides in yeast using a haploid mating strategy. US Patent 8,268,582.
|Ilya Tolstorukov, PhD|
|Location:||Building 535, Room 100|