By SDS-PAGE was around 26.7 kDa (Figure two). The molecular weight obtained by
By SDS-PAGE was roughly 26.7 kDa (Figure 2). The molecular weight obtained by SPSepharose and Sephacryl S-200 column chromatography was also around 26.7 kDa (Figure two).M 55.six 42.7 34.6 27.0 20.0 14.three six.Purified proteaseFigure two: SDS-PAGE in the purified protease. M: normal protein markers; lane 1: crude enzyme; lane two: ammonium sulphateprecipitated enzyme; lane 3: purified enzyme on SP-Sepharose (cation exchange); lane four: purified enzyme on Sephacryl S-200 (gel filtration).three.2. Optimum Temperature and Thermal Stability in the Purified Protease. The purified protease from red pitaya peel was active and stable all through a wide temperature range (20 C to 75 C). The temperature for the maximum protease Abl Storage & Stability activity was 70 C. At both 80 and 90 C, the protease was fairly active, with just about 60 and 35 activity, respectively. Consequently, theNaCl concentration (molarity)100 90 80 70 60 50 40 30 20 10500 450 400 350 300 250 200 150 100 50Serine protease (UmL)Serine protease (UmL)Absorbance at 280 nmBioMed Study International120 Relative activity ( ) 100 80 60 40 20 0 0 20 40 60 80 Temperature ( C)(a)120 Residual activity ( ) 100 80 60 40 20 0 -20 0 20 40 60 80 Temperature ( C)(b)120 Residual activity ( ) Relative activity ( ) one hundred 80 60 40 20 0 0 2(c)120 one hundred 80 60 40 20 0 0 two(d)six pH6 pHFigure 3: The optimum temperature (a), thermal stability (b), optimum pH (c), and pH stability (d) of purified thermoalkaline protease have been investigated.final results reveal that the optimum temperature for the enzyme is 70 C (Figure 3(a)). Evaluation of your thermal stability on the protease showed that the enzyme retained far more than 90 of its activity in the array of 20 to 80 C, however the enzyme activity was drastically ( 0.05) decreased at temperature above 80 C. The residual activity from the purified enzyme at 80 C was 23 , but above that temperature no detectable enzyme activity could possibly be determined (Figure 3(b)). This phenomenon could possibly be due to the denaturation from the enzyme at a heightened temperature. You will discover some reports in agreement with this study for isolated protease from some plant sources [19]. For that reason, the purified protease from pitaya peel showed the high thermostability. It needs to be pointed out that thermostability on the enzyme is one of the very good qualities of the protease. In addition, thermostable enzyme can decrease the risk of contaminants at higher temperature in industries and also cost of external cooling and also the elevated substrate solubility, allowing for larger concentrations of low solubility materials plus a decrease viscosity of liquids and it may also be JAK MedChemExpress useful in mixing. three.three. Impact of pH on Activity and Stability with the Purified Protease. Inside the pH activity experiments, the protease was observed to become around 75 active within the pH selection of 7.0 to 9.0 with 100 activity at pH 8.0. At pH levels of 3.0 and ten.0, the protease activity was lowered to 30 and 22 , respectively. The protease was as a result steady (3000 of maximum activity) throughout the whole pH range that was studied. The enzyme exhibited the highest stability (85 ) inside the pH variety four.0 to 10, with one hundred stability at pH eight.0 (Figure three(c)). The residual activity sharply decreased at pH levels above 10.0, with 33 on the initial activity with the enzymeobservable at a pH of 11.0 (Figure three(d)). The remarkable activity and stability more than a wide pH range reveal the highly alkaline nature of this protease, which tends to make it suitable for applications in alkaline environments a.
