Russian Journal of Bioorganic Chemistry

, Volume 44, Issue 1, pp 32–40 | Cite as

Biotechnological Method for Production of Recombinant Peptide Analgesic (Purotoxin-1) from Geolycosa sp. Spider Poison

  • R. S. Esipov
  • V. N. Stepanenko
  • I. O. Zvereva
  • D. A. Makarov
  • M. A. Kostromina
  • T. I. Kostromina
  • T. I. Muravyova
  • A. I. Miroshnikov
  • E. V. Grishin


Severe chronic and sometimes incurable diseases are frequently accompanied by a pain syndrome. Purotoxin-1, isolated from the poison of the Central Asian Geolycosa sp. spider and selectively inhibiting the purinergic P2X3 receptor (which is considered as a target for the control of the pain states), is one potentially highly effective drug with an analgesic effect. To produce the recombinant purotoxin-1, we created four genetically engineered constructions with different carrier proteins for the expression in E. coli: pTRX-PT1, pCBD-PT1, pGyrA-PT1, pDnaB-PT1. The construction with mini-intein DnaB from the Synechocystis sp. was the most efficient. Using the E. coli C3030/pDnaB-PT1 producer strain, the laboratory method, based on which a pilot technology of recombinant purotoxin-1 production was developed as a result of optimization and scaling, was created. Six grams of recombinant PT1 preparation with the confirmed pharmacological purity was developed for preclinical trials.


production of recombinant proteins intein analgesic peptide toxin 



adenosine triphosphate


chitinbinding domain


gyrase A


ethylenediaminetetraacetic acid




2-(N-morpholino)ethanesulfonic acid


phenylmethylsulfonyl fluoride






part per million


reversed-phase highperformance liquid chromatography


endotoxin unit


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Copyright information

© Pleiades Publishing, Ltd. 2018

Authors and Affiliations

  • R. S. Esipov
    • 1
  • V. N. Stepanenko
    • 1
  • I. O. Zvereva
    • 1
  • D. A. Makarov
    • 1
  • M. A. Kostromina
    • 1
  • T. I. Kostromina
    • 1
  • T. I. Muravyova
    • 1
  • A. I. Miroshnikov
    • 1
  • E. V. Grishin
    • 1
  1. 1.Shemyakin–Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesMoscowRussia

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