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Maximum protease activity (52,230 ± 1601 U g−1 DM) was observed at the 14th day of SSF.
By using this medium, an experimental maximum protease activity of 280.72 U/ml in 7 days cultivation verified the applied methodology.
In 5 l stirred-tank bioreactor, a maximum protease activity of 238.77 U/ml was attained in 6 days of cultivation when aeration rate and agitation speed were controlled at 0.6 vvm and 150 rpm, respectively.
On the other hand, in 30 l airlift bioreactor the maximum protease activity of 283.84 U/ml was found with the controlled aeration rate of 0.9 vvm operated in a bubble column mode.
Central composite design experiment in the following showed that the maximum protease activity (522 U/ml) was achieved in 0.0089 mM IPTG for 24 h at 30 °C, an OD600 of 2, 0.5% of peptone and yeast extract, and a Ca2+ ion concentration of 1.3 mM.
The optimized values obtained by the statistical analysis showed that soybean meal at 3.0 g/l, maltose50 between the ranges of 30 and 40 g/l, tween80 at 0.35 g/l and an initial pH of 8.0 gives maximum protease activity.
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The AS- protease activity was maximum at pH 10.5 (A) and at temperature 42°C (B) and these values were taken as 100% for comparison.
These results agree with the protease activity from Euphorbia milii where the maximum activity was recorded at pH 8.0, and the residual enzyme activity markedly decreased at pH levels above 10.0 [ 20].
Linear mixed model with REML was used to analyse the changes in maximum growth rate (log-transformed), biomass yield, motility (log-transformed), protease activity, and bacterial load in the hosts between the evolutionary treatments (within-host and outside-host).
However, Dastager et al. [ 13] showed that the protease activity measured in the cell-free supernatant fluid of Streptomyces gulbargensis sp. Nov. was maximum (121.8 U/mL) after 48 h of growth.
The maximum antioxidant activities were observed from 60 to 240 min of hydrolysis, where the degree of hydrolysis and residual protease activity were approximately 44.0 and 50.0%, respectively.
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