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Notes:
Volume 8
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X
Euro Biotechnology 2018
October 11-12, 2018
October 11-12, 2018 | Moscow, Russia
21
st
European
Biotechnology Congress
conferenceseries
.com
Page 41
Production strategies of thyroid peroxidase from
Branchiostoma belcheri
in
Escherichia coli
Monika Chovanova
1
, Gabriel Zorkocy
1
, Eva Struharnanska
1
, Marcel Zamocky
2
and
Stanislav Stuchlik
1
1
Comenius University in Bratislava, Slovakia
2
Slovak Academy of Sciences, Slovakia
P
eroxidases are catalytic enzymes that reduce hydrogen peroxide to oxygen and water and also oxidize a various
substrates. They are widely used in various branches of biotechnological industry as they are also applicated in
variety of high potential biotransformation reactions. Problem of using this enzyme is that it is usually difficult to obtain
an enough yield of enzyme or for their high-cost production. The aim of our research is to produce thyroid peroxidase
from
Branchiostoma belcheri
, for the structural studies and also because of its possible use in biotransformations. The
BbePOX1 gene sequence was prepared synthetically, codon-optimized for expression in
Escherichia coli
and cloned into
the pET-21a vector. The aims of the project are to select the optimal conditions for the production of thyroid peroxidase
(e.g. expression vector selection, expression and purification conditions procedures, etc.). To optimize expression from
vector and protein purification point of view we choose vectors for the high efficient expression of peroxidase (vector
with T5 promoter and vector with fusion partner). To achieve the highest yield of protein with the highest degree of
solubility and enzyme activity we have also focused on the optimal temperature of production, selection of suitable host
strain, concentration of inductor, selection of cultivate media and fermentation conditions. After isolation of a sufficient
amount of active recombinant thyroid peroxidase, we will be able to study its structure in the following steps, as well as
potentially used it in oxidoreduction biotransformation reactions.
Biography
Monika Chovanova has completed her Master´s degree in Biotechnology at the Comenius University in Bratislava, Faculty of Natural Sciences,
and Department of Molecular Biology. Theme of her diploma thesis was Heterologous expression and solubilization of synthetic peroxidase gene
Mag2C8 from
Magnaporthe oryzae
in
Escherichia coli
. After graduation in 2017, she began the PhD study in the same department in the theme:
Preparation of producers of biologically active substances.
monikachovanova2@gmail.comMonika Chovanova et al., J Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C5-101