Figures & data
Figure 1. (A) Effect of temperature on Bidhi (-□-) and Kahli (-▪-) amylase activity. (B) Thermal inactivation of Bidhi and Kahli amylase at 80°C. The enzyme activity was tested at various temperatures using soluble potato starch as substrate at pH 6.5 and 7 for both extracts, respectively. Residual activity was determined from 0 to 120 min. Values are means of three independent experiments. The initial activity before pre-incubation was taken as 100%. Standard deviations were <5%.
Figure 2. Effect of pH on Kahli (-▪-) and Bidhi (-□-) amylase activity (A) and stability (B). The enzyme activity was tested at various pHs using soluble potato starch as substrate at 45°C. The pH stability of both amylases was determined by incubating each enzyme in different buffers for 1 h at 4°C and the residual activity was measured at pH 6.5 and 7 for Kahli and Bidhi amylase, respectively, at 45°C. Buffer solutions used for pH activity and stability are mentioned in Materials and Methods section. The initial activity before pre-incubation was taken as 100%. Standard deviations were < 5%.
Figure 3. Effect of increasing Zn2+ (□), Cu2+ (☆), Fe2+ (▵), Mg2+ (▾) and Ca2+ (○) ion concentrations on Kahli (A), and Bidhi (B) amylase activities was determined after 5 min incubation in potato starch (1%) as substrate at pH 6.5 and 7, respectively, and 45°C. Each data point represents the mean of three independent assays (the standard errors were less than 5% of the means).
Figure 4. Effect of inhibitors on Kahli (A) and Bidhi (B) amylase activities was determined using potato starch (1%) as substrate at pH 6.5 and 7, respectively, and 45°C. Each data point represents the mean of three independent assays (the standard errors were less than 5% of the means).
Figure 5. Compatibility of amylase activities from Bidhi (A), (C) and Kahli (B), (D) with commercial detergents (-▪- Omino Bianco®, -○-OMO®, -★- Ariel®, -▿- Nadif®, -♦- Dixan®). The activities are expressed as a percentage of the activity levels in the absence of detergents (100%).
Table 1. Hydrolyzed products in various time courses (0–60 min) of digestion by Kahli and amylase.
Table 2. Hydrolyzed products in various time courses (0–60 min) of digestion by Bidhi amylase.
Figure 6. TLC analyses of reaction products of Bidhi (A) and Kahli (B) amylase from the left to right; standard oligosaccharides (lane 1), saccharose (lane 2), reaction products for 0 min (lane 3), 5 min (lane 4), 15 min (lane 5), 30 min (lane 6), 45 min (lane 7) and 60 min (lane 8). Lane 1 indicates the standard oligosaccharides (G1, glucose; G2, maltose; G3, maltotriose; G4, maltotetraose; G5, maltopentose).
Figure 7. High-performance liquid chromatography of starch hydrolysate produced during serial time of incubation (0, 15, 30, 45 and 60 min) by Bidhi (2, 3, 4, 5, 6) and Kahli (7, 8, 9, 10, 11) amylase. Standard (1): saccharose, glucose and fructose.
