Group of fungal genetic engineering

 eldarov_1 Michael A. Eldarov
Ph.D.
Head of the Group
INB, room 401
Телефон +7 (499) 135-62-19
E-Mail eldarov@biengi.ac.ru

Our group is interested in rather diverse aspects of regulation of primary and secondary metabolism in yeast and fungi. The main topic is the study of molecular control of cephalosporin C production  (CPC) in filamentous fungus Acremonium chrysogenum with the focus of biogenic polyamines as inducer of antibiotic production, cell survival and stress resistance in this organism. We have developed agrobacterim-mediated transformation system for industrial A.chrysogenum strains,  determined  the dynamics of gene expression of key cephalosporin C biosynthesis and transport genes in the course of CPC biosynthesis,  specified the functions of A.chrysogenum  CefT protein, definied differences  in polyphosphate metabolism and cell-wall structures in laboratory and industrial A.chrysogenum strains.

We also collaborate with our partners in the field of enzymology of phosphorus metabolism in yeast and fungi (G.K.Skryabin Institute, Puschino), protein engineering of the enzymes of biotransformation of beta-lactam antibiotics and sperm-specific glyceraldehyde-phosphate dehydrogenase (Belozersky Institute, MSU), genomics of Wine Yeast (Magarach Institute of Viticulture and Wine-Making).

Particularly, we have developed efficient expression systems for production of recombinant PPX and PPN – yeast enzymes of polyphosphate metabolism, detailed their enzymatic properties.  Structural basis of substrate specificity of and quaternary structure formation were determined for Brevundimonas diminuta  glutaryl-acylase and novel more efficient analogs of this enzyme were prepared.

In collaboration with the group of genomics of microorganisms  we have deciphered mitochondrial genomes of several yeast and fungi, elucidated genome and transcriptome of methylotrophic yeast H.polymorpha and now try to apply these knowledge for further improvements  of H.polymorpha for metabolic engineering studies.

 

Selected publications:

  1. Zakirova SA, Mikhaĭlova TV, Él’darov MA. [Peculiarities of the Brevundimonas diminuta Gl7ACA-Acylase quaternary structure formation and obtaining stable enzyme analogues]. Prikl Biokhim Mikrobiol. 2013 Nov-Dec;49(6):554-60. Russian.
  2. Dumina MV, Zhgun AA, Kerpichnikov IV, Domracheva AG, Novak MI, Valiakhmetov AIa, Knorre DA, Severin FF, Él’darov MA, Bartoshevich IuÉ. [Functional characteristic of the CefT transporter of the MFS family involved in the transportation of beta-lactam antibiotics in Acremonium chrysogenum and Saccharomyces cerevisiae]. Prikl Biokhim Mikrobiol. 2013 Jul-Aug;49(4):372-81.
  3. Redo VA, Novikova EK, Él’darov MA. [Expression of modified oxidase of D-aminoacids of Trigonopsis variabilis in methylotrophic yeasts Pichia pastoris]. Prikl Biokhim Mikrobiol. 2011 Jan-Feb;47(1):39-45. Russian.
  4. Elkina YL, Kuravsky ML, El’darov MA, Stogov SV, Muronetz VI, Schmalhausen EV. Recombinant human sperm-specific glyceraldehyde-3-phosphate dehydrogenase: structural basis for enhanced stability. Biochim Biophys Acta. 2010 Dec;1804(12):2207-12.
  5. Kelebina TS, Seliakh IO, Gorkovskiĭ AA, Bezsonov EE, El’darov MA, Novak MI, Domracheva AG, Bartoshevich IuE. [Structure peculiarities of cell walls of Acremonium chrysogenum–an autotroph of cephalosporin C]. Prikl Biokhim Mikrobiol. 2010 Nov-Dec;46(6):666-71.
  6. Valiakhmetov AIa, Trilisenko LV, Vagabov VM, Bartoshevich IuE, Kulaev IS, Novak MI, Domracheva AG, El’darov MA, Skriabin KG. [The concentration dynamics of inorganic polyphosphates during the cephalosporin C synthesis by Acremonium chrysogenum]. Prikl Biokhim Mikrobiol. 2010 Mar-Apr;46(2):198-204. Russian.
  7. Meshcheriakova IuA, El’darov MA, Migunov AI, Stepanova LA, Repko IA, Kiselev OI, Lomonosov DP, Skriabin KG. [Cowpea mosaic virus chimeric particles bearing ectodomain of matrix protein 2 (M2E) of influenza A virus: production and characteristics]. Mol Biol (Mosk). 2009 Jul-Aug;43(4):741-50.
  8. Zhgun AA, Ivanova MA, Domracheva AG, Novak MI, El’darov MA, Skriabin KG, Bartoshevich IuE. [Genetic transformation of the mycelium fungi Acremonium chrysogenum]. Prikl Biokhim Mikrobiol. 2008 Nov-Dec;44(6):663-70.
  9. Zhgun AA, El’darov MA, Solodar’ LI, Sokolov NN, Archakov AI, Skriabin KG. [Heterologous expression of eukaryotic CYP450. 1. Heterologous expression of cytochrome P450 2B4 using groups with various affinity in E. coli]. Vopr Med Khim. 2001 Jul-Aug;47(4):382-92.
  10. Trilisenko LV, Andreeva NA, Eldarov MA, Dumina MV, Kulakovskaya TV. Polyphosphates and Polyphosphatase Activity in the Yeast Saccharomyces cerevisiae during Overexpression of the DDP1 Gene. Biochemistry (Mosc). 2015 Oct;80(10):1312-7.
  11. Eldarov MA, Sklyarenko AV, Dumina MV, Medvedeva NV, Jgoun AA, Satarova JE, Sidorenko AI, Emperian AS, Yarotsky SV. [Recombinant cephalosporin-acid synthesase: optimisation of expression in E.coli cells, immobilisation and application for biocatalytic cefazolin synthesis]. Biomed Khim. 2015 Sep-Oct;61(5):646-51.
  12. Sokolov NN, Eldarov MA, Pokrovskaya MV, Aleksandrova SS, Abakumova OY, Podobed OV, Melik-Nubarov NS, Kudryashova EV, Grishin DV, Archakov AI. [Bacterial recombinant L-asparaginases: properties, structure and anti-proliferative activity]. Biomed Khim. 2015 May-Jun;61(3):312-24.
  13. Andreeva N, Trilisenko L, Kulakovskaya T, Dumina M, Eldarov M. Purification and properties of recombinant exopolyphosphatase PPN1 and effects of its overexpression on polyphosphate in Saccharomyces cerevisiae. J Biosci Bioeng. 2015 Jan;119(1):52-6.
  14. Eldarov MA, Mardanov AV, Beletsky AV, Dumina MV, Ravin NV, Skryabin KG. Complete mitochondrial genome of the cephalosporin-producing fungus Acremonium chrysogenum. Mitochondrial DNA. 2015;26(6):943-4.
  15. Andreeva N, Trilisenko L, Eldarov M, Kulakovskaya T. Polyphosphatase PPN1 of Saccharomyces cerevisiae: switching of exopolyphosphatase and endopolyphosphatase activities. PLoS One. 2015 Mar 5;10(3):e0119594.
  16. Lichko LP, Eldarov MA, Dumina MV, Kulakovskaya TV. PPX1 gene overexpression has no influence on polyphosphates in Saccharomyces cerevisiae. Biochemistry (Mosc). 2014 Nov;79(11):1211-5.
  17. Dumina MV, Zhgun AA, Novak MI, Domratcheva AG, Petukhov DV, Dzhavakhiya VV, Eldarov MA, Bartoshevitch IuE. Comparative gene expression profiling reveals key changes in expression levels of cephalosporin C biosynthesis and transport genes between low and high-producing strains of Acremonium chrysogenum. World J Microbiol Biotechnol. 2014 Nov;30(11):2933-41.
  18. Kuravsky M, Barinova K, Marakhovskaya A, Eldarov M, Semenyuk P, Muronetz V, Schmalhausen E. Sperm-specific glyceraldehyde-3-phosphate dehydrogenase is stabilized by additional proline residues and an interdomain salt bridge. Biochim Biophys Acta. 2014 Oct;1844(10):1820-6.
  19. Ravin NV, Eldarov MA, Kadnikov VV, Beletsky AV, Schneider J, Mardanova ES, Smekalova EM, Zvereva MI, Dontsova OA, Mardanov AV, Skryabin KG. Genome sequence and analysis of methylotrophic yeast Hansenula polymorpha DL1. BMC Genomics. 2013 Nov 27;14:837.
  20. Eldarov MA, Baranov MV, Dumina MV, Shgun AA, Andreeva NA, Trilisenko LV, Kulakovskaya TV, Ryasanova LP, Kulaev IS. Polyphosphates and exopolyphosphatase activities in the yeast Saccharomyces cerevisiae under overexpression of homologous and heterologous PPN1 genes. Biochemistry (Mosc). 2013 Aug;78(8):946-53.
  21. Thuenemann EC, Lenzi P, Love AJ, Taliansky M, Bécares M, Zuñiga S, Enjuanes L, Zahmanova GG, Minkov IN, Matić S, Noris E, Meyers A, Hattingh A, Rybicki EP, Kiselev OI, Ravin NV, Eldarov MA, Skryabin KG, Lomonossoff GP. The use of transient expression systems for the rapid production of virus-like particles in plants. Curr Pharm Des. 2013;19(31):5564-73.
  22. Eldarov MA, Mardanov AV, Beletsky AV, Dzhavakhiya VV, Ravin NV, Skryabin KG. Complete mitochondrial genome of compactin-producing fungus Penicillium solitum and comparative analysis of Trichocomaceae mitochondrial genomes. FEMS Microbiol Lett. 2012 Apr;329(1):9-17.
  23. Eldarov MA, Mardanov AV, Beletsky AV, Ravin NV, Skryabin KG. Complete sequence and analysis of the mitochondrial genome of the methylotrophic yeast Hansenula polymorpha DL-1. FEMS Yeast Res. 2011 Sep;11(6):464-72.
  24. Andrianova EP, Krementsugskaia SR, Lugovskaia NN, Mayorova TK, Borisov VV, Eldarov MA, Ravin NV, Folimonov AS, Skryabin KG. Foot and mouth disease virus polyepitope protein produced in bacteria and plants induces protective immunity in guinea pigs. Biochemistry (Mosc). 2011 Mar;76(3):339-46.
  25. Khatuntseva SA, Eldarov MA, Redo VA, Skryabin KG. Purification and immobilization of recombinant variants of Brevundimonas diminuta glutaryl-7-aminocephalosporanic acid acylase expressed in Escherichia coli cells. J Biotechnol. 2008 Jan 1;133(1):123-6.