Expression of DNA-Encoded Antidote to Organophosphorus Toxins in the Methylotrophic Yeast Pichia Pastoris
- Authors: Terekhov S.S.1, Bobik T.V.1, Mokrushina Y.A.1, Stepanova A.V.1, Aleksandrova N.M.2, Smirnov I.V.1,2, Belogurov A.A.1,2, Ponomarenko N.A.1, Gabibov A.G.1,2
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Affiliations:
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- Kazan (Volga Region) Federal University
- Issue: Vol 52, No 2 (2016)
- Pages: 162-169
- Section: Article
- URL: https://journal-vniispk.ru/0003-6838/article/view/151868
- DOI: https://doi.org/10.1134/S0003683816020162
- ID: 151868
Cite item
Abstract
A platform for the cloning and expression of active human butyrylcholinesterase (BuChE) in the yeast Pichia рastoris is first presented. Genetic constructs for BuChE gene expression, separately and in conjunction with a proline-rich peptide called proline-rich attachment domain (PRAD), are based on the vector pPICZαA. It is shown that the highest level of production is achieved in the expression of a BuChE gene without PRAD pPICZαA. It is found that one can obtain up to 125 mg of active enzyme from 1 L of culture medium at an optimal pH environment (pH 7.6), an optical seed culture density of 3 o.u., and an optimum methanol addition mode of (0.5% methanol in the first day and 0.2% thereafter from the second day).
About the authors
S. S. Terekhov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997
T. V. Bobik
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997
Yu. A. Mokrushina
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997
A. V. Stepanova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997
N. M. Aleksandrova
Kazan (Volga Region) Federal University
Email: gabibov@mx.ibch.ru
Russian Federation, Kazan, 420000
I. V. Smirnov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Kazan (Volga Region) Federal University
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997; Kazan, 420000
A. A. Belogurov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Kazan (Volga Region) Federal University
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997; Kazan, 420000
N. A. Ponomarenko
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997
A. G. Gabibov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry; Kazan (Volga Region) Federal University
Author for correspondence.
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow, 117997; Kazan, 420000
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