Faculty & Staff

Ilya Tolstorukov, PhD, DSc

Dr. Ilya Tolstorukov joined the Keck Graduate Institute (KGI) of Applied Life Sciences in May 2001. Prior to this, he conducted extensive research in yeast genetics, molecular biology, and biotechnology. His career includes roles as a researcher and Head of the Genetics of Methylotrophic Yeasts Section at the State Research Institute of Genetics and Selection of Industrial Microorganisms (VNIIGenetika) in Moscow, Russia. He has also been a Visiting Scientist at the Laboratory of Eukaryotic Gene Expression at the Frederick Cancer Research Facility of NIH (Frederick, MD), the Laboratory for Electron Microscopy at the Biological Centre of the University of Groningen (Haren, Netherlands), and a Visiting Professor at Osaka University (Japan).

Dr. Tolstorukov is a co-founder of BioGrammatics, Inc. (Carlsbad, CA), a contract research organization providing services, consulting, and reagents for protein expression using the Pichia pastoris platform.

  • G.I.Naumov, I.I.Tolstorukov. 1973. Comparative genetics of yeasts. Communication 10. Re-identification of mutators of mating types in Saccharomyces. Genetika (Soviet Genetics), v.9, No.1, p.82-91.
  • I.I.Tolstorukov, T.A.Dutova, S.V.Benevolensky, Ya.O.Soom. 1977. Hybridization and genetic analysis of methanol-utilizing yeast Pichia pinus. Genetika (Soviet Genetics), v.13, N.2, pp.322-329.
  • I.I.Tolstorukov, S.V.Benevolensky. 1982. Genetic control of cell type and complex organization of the mating type locus in the yeast Pichia pinus. Current Genetics, v.5, p.137-142.
  • M.A.Neystat, V.V.Alenin, I.I.Tolstorukov. 1986. Transformation of the yeasts Hansenula polymorpha, Pichia guilliermondii, Williopsis saturnus by plasmid containing the gene ADE2 from Saccharomyces cerevisiae. Molecularnaya Genetika, Microbiologia and Virusologiya, N 12, p.19-23.
  • A.A.Sibirny, V.I.Titorenko, B.D.Efremov, I.I.Tolstorukov. 1987. Multiplicity of mechanisms of carbon catabolite repression involved in the synthesis of alcohol oxidase in the methylotrophic yeast Pichia pinus. Yeast, v.3, N.4, p.233-241.
  • I.I.Tolstorukov. 1988. Genetic of methylotrophic yeast Pichia pinus. D.Sc. Thesis, Moscow, VNIIGenetika, K.L.Lahthev, Y.D.Semenova, I.I.Tolstorukov, I. Van der Klei, M.Veenhuis. 2002. Isolation and properties of genetically defined strains of the methylotrophic yeast Hansenula polymorpha CBS4732. Arch. Microbiol., V. 177, 150-158.
  • G.P.Lin-Cereghino, L.Godfrey, B.J.de la Cruz, S.Johnson, S.Khuingsthiene, I.Tolstorukov, M.Yan, J.Lin-Cereghino, M.Veenhuis, S.Subramani, J.M.Cregg. 2006. Mxr1p: a key regulator of the methanol-utilization pathway and peroxisomal genes in Pichia pastoris. Mol. and Cell Biol., V.26, No. 3, 883-897.
  • J. Cregg, J. Latham, M. Litton, R.Schatzman, I. Tolstorukov. Methods of synthesizing heteromultimeric polypeptides in yeast using a haploid mating strategy. US Patent 7,927,863. (Priority: May 8, 2006)
  • J.M.Cregg, I.I.Tolstorukov, A.Kusari, J.Sunga, K.Madden, T.Chappell. 2009. Expression in the yeast Pichia pastoris. Review. Methods Enzymol. 463, 169-189.
  • I. Tolstorukov & J. Cregg. Yeast promoters from Pichia pastoris. US Patent 10,619,164 (priority March 8, 2013)
  • L. Sturmberger, T.G. Chappell, M. Geier, F. Krainer, K.J. Day, U. Vide, S. Trstenjak, A. Schiefer, T. Richardson, L. Soriaga, B. Darnhofer, R. Birner-Grünberger, B.S. Glick, I. Tolstorukov, J. Cregg, K. Madden and A. Glieder. (2016). Refined Pichia pastoris reference sequence. J Biotechnol. S0168-1656(16)30203-6. doi: 10.1016/j.jbiotec.2016.04.023. PMID: 27084056
  • I.Tolstorukov. T.G.Chappell, J.M.Cregg, 2025. Chapter 8. Classical Genetics. Methods in Mol. Biol. “Pichia pastoris: Methods and Protocols” (Ed. A.Glieder, K.Kovar, X.G.Ortega), Springer Nature. (In press)

Dr. Tolstorukov’s expertise lies in engineering yeast cell lines to produce valuable bioproducts such as antibodies, enzymes for agriculture and medicine, and polysaccharides. His research also focuses on mating and the regulation of life cycles in yeast.

His early work (Ph.D. thesis) involved unraveling complex genes controlling mating types and homothallic behavior in Saccharomyces cerevisiae. Although initially published in Russian journals, his findings gained international recognition and were integral to the development of the "Cassette Model of Mating Type Switching" by leading yeast geneticists.

As a postdoctoral fellow, Dr. Tolstorukov shifted his focus to methylotrophic yeasts, a group of microorganisms with significant biotechnological potential. His group was the first to develop genetic and molecular tools for these non-conventional yeasts, leading to a series of influential publications, Ph.D. theses, and his D.Sc. dissertation. This foundational work on methylotrophic yeasts enabled the development of Pichia pastoris, Pichia methanolica, and Hansenula polymorpha protein expression platforms that are widely used today.

Achievements at KGI:

At KGI, Dr. Tolstorukov collaborated with founding professor James Cregg to advance the application of the Pichia pastoris system for protein expression. One of his first projects involved developing Pichia pastoris as a host for expressing complex proteins like monoclonal antibodies. This proprietary technology, patented by KGI and licensed to Alder Biopharmaceuticals, contributed to the production of therapeutic monoclonal antibodies. In 2020, Lundbeck launched Viepty, the first Pichia-derived monoclonal antibody biologic for the treatment of migraines.

During his tenure, Dr. Tolstorukov has developed numerous Pichia pastoris strains used in manufacturing antibodies, enzymes, and other vital proteins, significantly advancing biotechnological research and applications.

  1. I.Tolstorukov, T.Prosvirova, et al. 1993. Strain of Candida utilis for production of single cell protein. Certificate of Invention (Russia), RU N 93019585.
  2. J.Cregg, J.Latham, M.Litton, R.Schatzman, I.Tolstorukov. 2011. Methods of synthesizing heteromultimeric polypeptides in yeast using a haploid mating strategy. US Patents 7927863, 8268582 and 8709756.
  3. J.Cregg, I.Tolstorukov. 2012. P. pastoris ADH promoter and use thereof to direct expression of proteins. US patent 8222386.
  4. J.Cregg, I.Tolstorukov. 2014. Novel P. pastoris promoters, and the use thereof to direct expression of proteins in yeast, preferably using a haploid mating strategy. US Patent 8785613.