Biomedical Engineering Reference
In-Depth Information
Chapter 4
O
PTIMIZATION OF THE
P
ROCESS OF
C
ULTIVATION OF
R
ECOMBINANT
E
SCHERICHIA
C
OLI
TBI
P
ROTEIN
P
RODUCING
S
TRAIN
N. I. Akulova,
*
T. A. Kashperova, E. A. Volosnikova
and L. R. Lebedev
Federal State Research Institution “State Research Center of Virology and Biotechnology
“Vector”, Institute of Medical Biotechnology, Berdsk, Novosibirsk Region, Russia
A
BSTRACT
The effect of cultivation conditions on the yield of TBI protein in the process of
biosynthesis of recombinant
E.coli
JM 103/pTBI
strain was studed. Cultivation
conditions were optimized by the composition of nutrient medium, the time of addition of
the protein biosynthesis inducer and the rate of aeration in the process of cultivation in
flasks and laboratory fermenters. It was shown that the yield of target protein increased in
an optimized medium by two-fold as compared to the control. The qualitative and
quantitative characteristics of TBI protein isolated from the optimized TB plus medium
were better as compared to the control.
Keywords:
Cultivation, recombinant strain, TBI protein, nalidixic acid, TB plus medium,
biosynthesis, biomass, fermentation
I
NTRODUCTION
The process of microbiological synthesis of TBI protein plays a key role in technology of
production of CombiHIVVAC vaccine developed by the FSRI SRC VB “Vector” [1, 2]. TBI
protein is generated in
E.coli
procaryotic cells. The level of gene expression of recombinant
TBI protein at the stage of biomass is determined the quality of the inoculate, the conditions
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