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conferenceseries
.com
Volume 8
Journal of Biotechnology & Biomaterials
ISSN: 2155-952X
Pharma Biotech 2018
December 10-11, 2018
December 10-11, 2018 | Rome, Italy
23
rd
International Conference on
Pharmaceutical Biotechnology
Optimization of fermentation conditions for extracellular production of a therapeutic protein drug
uricase, by
Aspergillus welwitschiae
using response surface methodology
Noura El-Ahmady El-Naggar
1
, S A Haroun
2
, Eman M El-Weshy
2
and
A A Sherief
2
1
Genetic Engineering and Biotechnology Research Institute, Egypt
2
Mansoura University, Egypt
U
ricase is commonly used in clinical analysis for the determination of urate in blood and other biological fluids. Microbial
uricases as protein drug have been found effective in the treatment of hyperuricemia and gout, as well as in prophylaxis
and treatment of tumor lysis hyperuricemia. A potential culture,
Aspergillus
sp. strain 1-4 displayed high extracellular
uricase activity. This uricolytic fungal isolate was identified as
Aspergillus welwitschiae
strain 1-4 on the basis of phenotypic
characteristics, together with ITS region sequence analysis. Sequencing product was deposited in the GenBank database under
accession number MG323529. The Plackett-Burman experimental design with 20 runs was applied for screening of fifteen
variables for their significances on uricase production by
Aspergillus welwitschiae
. Incubation time was the most significant
variable affecting uricase production followed by yeast extract and inoculum size with significant P-values of 0.0002, 0.0083 and
0.0118; respectively. These variables were chosen for optimization studies using central composite design. It was found that, the
maximum uricase production (59.01 U/mL) by
Aspergillus welwitschiae
is achieved at the following fermentation conditions
g/L: sucrose 30, uric acid 3, peptone 2, yeast extract 2, NaNO
3
2, K
2
HPO
4
1, MgSO
4
.7H
2
O 0.2, NaCl 0.2 and FeSO
4
.7H
2
O 0.03”,
incubation time 7 days, temperature 35, pH 6, inoculum size 4 mL/50 mL medium, inoculum age 72 h and medium volume
50 mL/250 mL conical flask. An overall 2.5-fold increase in uricase production by
Aspergillus welwitschiae
was achieved in
the medium after statistical optimization as compared with the unoptimized basal medium (23.58 U/mL) before applying
Plackett-Burman.
nouraelahmady@yahoo.comJ Biotechnol Biomater 2018, Volume 8
DOI: 10.4172/2155-952X-C8-110