By SDS-PAGE was around 26.7 kDa (Figure 2). The molecular weight obtained by
By SDS-PAGE was around 26.7 kDa (Figure two). The molecular weight obtained by SPSepharose and Sephacryl S-200 column chromatography was also approximately 26.7 kDa (Figure two).M 55.6 42.7 34.six 27.0 20.0 14.three 6.Purified proteaseFigure two: SDS-PAGE of the purified protease. M: typical protein markers; lane 1: crude enzyme; lane 2: ammonium sulphateprecipitated enzyme; lane 3: purified enzyme on SP-Sepharose (cation exchange); lane four: purified enzyme on Sephacryl S-200 (gel filtration).3.two. HSPA5 supplier 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 variety (20 C to 75 C). The temperature for the maximum protease activity was 70 C. At both 80 and 90 C, the protease was very active, with virtually 60 and 35 activity, respectively. For that reason, theNaCl concentration (molarity)100 90 80 70 60 50 40 30 20 10500 450 400 350 300 250 200 150 one hundred 50Serine protease (UmL)Serine protease (UmL)Absorbance at 280 nmBioMed Study International120 Relative activity ( ) one hundred 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 two(c)120 one hundred 80 60 40 20 0 0 two(d)6 pH6 pHFigure 3: The optimum temperature (a), thermal stability (b), optimum pH (c), and pH stability (d) of purified thermoalkaline protease had been investigated.outcomes reveal that the optimum temperature for the enzyme is 70 C (Figure 3(a)). Evaluation of the thermal stability of the protease showed that the enzyme retained additional than 90 of its activity inside the array of 20 to 80 C, but the enzyme activity was considerably ( 0.05) decreased at temperature above 80 C. The residual activity in the purified enzyme at 80 C was 23 , but above that temperature no detectable enzyme activity might be determined (Figure three(b)). This phenomenon could be as a result of denaturation on the enzyme at a heightened temperature. You will find some reports in agreement with this study for isolated protease from some plant sources [19]. Consequently, the purified protease from pitaya peel showed the CXCR7 Compound higher thermostability. It ought to be talked about that thermostability on the enzyme is among the great traits of your protease. Also, thermostable enzyme can reduce the danger of contaminants at higher temperature in industries and also expense of external cooling and also the enhanced substrate solubility, allowing for larger concentrations of low solubility materials as well as a reduce viscosity of liquids and it might also be helpful in mixing. three.3. Impact of pH on Activity and Stability with the Purified Protease. Inside the pH activity experiments, the protease was observed to become approximately 75 active within the pH selection of 7.0 to 9.0 with one hundred activity at pH eight.0. At pH levels of 3.0 and 10.0, the protease activity was lowered to 30 and 22 , respectively. The protease was as a result stable (3000 of maximum activity) all through the complete pH range that was studied. The enzyme exhibited the highest stability (85 ) inside the pH variety 4.0 to 10, with 100 stability at pH 8.0 (Figure 3(c)). The residual activity sharply decreased at pH levels above 10.0, with 33 in the initial activity with the enzymeobservable at a pH of 11.0 (Figure 3(d)). The remarkable activity and stability more than a wide pH range reveal the extremely alkaline nature of this protease, which makes it appropriate for applications in alkaline environments a.
