Optimization of the Expression of Nitrilase from Alcaligenes denitrificans in Rhodococcus rhodochrous to Improve the Efficiency of Biocatalytic Synthesis of Ammonium Acrylate
- Authors: Lavrov K.V.1, Voronin S.P.2, Minasyan R.A.2, Glinskii S.A.2, Epremyan A.S.3, Kalinina T.I.1, Novikov A.D.1, Shemyakina A.O.1, Grechishnikova E.G.1, Yanenko A.S.1
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Affiliations:
- State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
- BIOAMIDE Closed Joint Stock Venture
- Scientific Production Center Armbiotechnology, National Academy of Sciences of Armenia
- Issue: Vol 55, No 9 (2019)
- Pages: 861-869
- Section: Producers, Biology, Selection, and Gene Engineering
- URL: https://journals.rcsi.science/0003-6838/article/view/153121
- DOI: https://doi.org/10.1134/S0003683819090035
- ID: 153121
Cite item
Abstract
A Rhodococcus rhodochrous strain, M33-2nit, has been constructed with two copies of the nitrilase gene from A. denitrificans В-9582 under the control of nitrile hydratase promoter from R. rhodochrous M8. The optimized cultivation of this strain made it possible to obtain the enzyme in a concentration of up to 17 g of dry cells/L with a specific activity of up to 7 U/mg cdw with a two-substrate culturing scheme in a fed-batch reactor with the sequential addition of glucose and acetate. The capacities of A. denitriificans В-9582 and R. rhodochrous M33-2nit cells to synthesize ammonium acrylate from acrylonitrile under conditions imitating industrial synthesis are compared. It is shown that R rhodochrous M33-2nit cells can synthesize ammonium acrylate under higher rates of acrylonitrile feeding than A. denitriificans В-9582 cells. The potential to obtain a highly concentrated solution of ammonium acrylate (450 g/L) with R. rhodochrous M33-2nit cells as a biocatalyst was demonstrated. The conversion of acrylonitrile to ammonium acrylate reached 99.5%.
About the authors
K. V. Lavrov
State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
Author for correspondence.
Email: lavrov.ko@gmail.com
Russian Federation, Moscow, 117545
S. P. Voronin
BIOAMIDE Closed Joint Stock Venture
Email: yanenko@genetika.ru
Russian Federation, Saratov, 410033
R. A. Minasyan
BIOAMIDE Closed Joint Stock Venture
Email: yanenko@genetika.ru
Russian Federation, Saratov, 410033
S. A. Glinskii
BIOAMIDE Closed Joint Stock Venture
Email: yanenko@genetika.ru
Russian Federation, Saratov, 410033
A. S. Epremyan
Scientific Production Center Armbiotechnology, National Academy of Sciences of Armenia
Email: yanenko@genetika.ru
Armenia, Yerevan, 0056
T. I. Kalinina
State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
Email: yanenko@genetika.ru
Russian Federation, Moscow, 117545
A. D. Novikov
State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
Email: yanenko@genetika.ru
Russian Federation, Moscow, 117545
A. O. Shemyakina
State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
Email: yanenko@genetika.ru
Russian Federation, Moscow, 117545
E. G. Grechishnikova
State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
Email: yanenko@genetika.ru
Russian Federation, Moscow, 117545
A. S. Yanenko
State Research Institute of Genetics and Selection of Industrial Microorganisms, Kurchatov Institute National Research Center (GosNIIgenetika, Kurchatov Institute NRC)
Author for correspondence.
Email: yanenko@genetika.ru
Russian Federation, Moscow, 117545
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