Table 3: Detected Lectin Samples Temperature (0C) pH Amount of lectin detected (mg/kg) Fermentation time 12 Hours 24 Hours 36 Hours Unripen banana 20 5.5 0.5 0.6 0.4 7.0 0.9 1.0 0.6 8.5 0.8 0.7 0.8 25 5.5 0.7 0.6 0.5 7.0 1.3 1.0 1.2 8.5 1.0 0.9 1.0 30 5.5 0.4 0.6 0.3 7.0 1.0 0.9 0.8
33 | P a g e 4.2. Carbohydrate Agglutination Assay of Banana Lectin 100 μl of the lectin were dropped on the KHB kit. Color line were observed after 15 min( agglutination was observed).Throughout purification of the lectin, KHB kit were used as indicator. Agglutination was observed from ripen banana sample that was socked in pH 7, in 24 hours and at a temperature of 25 o C when dropped into KHB kit. This indicates that the purified samples had a lectin and this protein binds to the carbohydrate fragment of glycoprotein (gp120) of the KHB kit. 8.5 0.5 0.6 0.7 Ripped banana 20 5.5 1.2 1.5 1.3 7.0 2.3 2.6 2.4 8.5 1.8 1.7 2.0 25 5.5 2.2 2.4 2.3 7.0 3.4 3.6 3.3 8.5 3.0 2.7 3.0 30 5.5 1.0 1.1 1.0 7.0 2.2 2.4 2.4 8.5 1.3 1.7 1.2
34 | P a g e Figure 6: Lectin activity on KHB kit 4.3. Model adequacy check In this experiment there were three kinds of factors (temperature, pH, and time) for two kinds of samples with one replicate and one response which was amount of lectin. Because of this, the overall experiments were become around 54. The result of each experiment was recorded in the table form. Finally each result was fed to design experiment in order to analyze every factor. The adequacy of the model was further checked with analysis of variance (ANOVA) as shown at the end of the appendix and the graph of the predicted values obtained using the developed correlation versus actual values is also shown in the next figure.
35 | P a g e Figure 7: predicted versus actual lectin activity This plot represents the line of perfect fit with points corresponding to zero error between predicted values and actual values and demonstrated that the regression model equation provided accurate description of the experimental data, in which all the points are close to the line of perfect fit. This result indicates that it was successful in capturing the correlation between the three variables to the lectin activity. 4.3.1. Effect of temperature Temperature is one of the important factors that affect the amount of lectin extracted. Because proteins are highly sensitive to high temperature protein denaturation may result. Temperature shows significant effect on the amount of lectin protein as shown in figure 8. Temperature has both positive effect and negative effect on lectin amount and as temperature increased the amount of lectin was also increased until 25 o C but after this temperature lectin amount was decreased as temperature increased. This is due to temperature has significant effect on lectin amount. The present study indicates amount of lectin increases with increasing temperature until DESIGN-EXPERT Plot Amount of lectin Actual Predicted Predicted vs. Actual 0.05 0.93 1.82 2.71 3.60 0.05 0.93 1.82 2.71 3.60
36 | P a g e optimum temperature conditions but further increasing of temperature would decrease the amount.
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- Fall '19