Partial Purification and Characterization of a Heat Stable α-Amylase from a Thermophilic Actinobacteria,Streptomycessp. MSC702

Abstract

A partial purification and biochemical characterization of the α-amylase from Streptomyces sp. MSC702 were carried out in this study. The optimum operational conditions for enzyme substrate reaction for amylolytic enzyme activity from the strain were evaluated. The optimum pH, temperature, and incubation period for assaying the enzyme were observed to be 5.0, 55°C, and 30 min, respectively. The extracellular extract was concentrated using ammonium sulfate precipitation. It was stable in the presence of metal ions (5 mM) such as K+, Co2+, and Mo2+, whereas Pb2+, Mn2+, Mg2+, Cu2+, Zn2+, Ba2+, Ca2+, Hg2+, Sn2+, Cr3+, Al3+, Ag+, and Fe2+ were found to have inhibitory effects. The enzyme activity was also unstable in the presence of 1% Triton X-100, 1% Tween 80, 5 mM sodium lauryl sulphate, 1% glycerol, 5 mM EDTA, and 5 mM denaturant urea. At temperature 60°C and pH 5.0, the enzyme stability was maximum. α-amylase retained 100% and 34.18% stability for 1 h and 4 h, respectively, at 60°C (pH 7.0). The enzyme exhibited a half-life of 195 min at 60°C temperature. The analysis of kinetic showed that the enzyme has K m of 2.4 mg/mL and V max of 21853.0 μmol/min/mg for soluble potato starch. The results indicate that the enzyme reflects their potentiality towards industrial utilization.

Production and optimization of α-amylase from a novel thermoalkalophilic Bacillus sonorensis GV2 isolated from mushroom compost

“…Ca 2+ and Mg 2+ are reported to enhance enzyme activity by Goyal et al (2005); Ashger et al (2007) and Kiran and Chandra, (2008). In accordance to the findings of Burhan et al (2003) and Singh et al (2014), Ni 2+ and Zn 2+ almost inhibited the enzyme production while in the medium. Ba 2+ , Fe 2+ and Co 2+ showed a negative effect on enzyme production lowering the titres considerably.…”

“…4), which is significantly higher than others. Similarly, optimum incubation temperature of 50°C for an enhanced amylase production has been reported in studies by Kunamneni et al (2005), Hernández, (2006), Asgher et al (2007);Aygan et al (2008); Zohra and Ahmad, (2012) and Singh et al (2014). A decrease in enzyme activity i.e.…”

Production and Characterization of Hydrocarbon Degrading Peroxidase from Rhizopus and Saccharomyces Species

“…However, Pb 2+ inhibited the activity of the enzyme. Singh et al (2014) have also reported an inhibitory effect by Hg 2+ , Pb 2+ and Cu 2+ and attributed this inhibition by these ions on the enzyme activity to the presence of indole amino acids in the enzyme and also a reflection of competition between exogenous and protein associated cations. In the presence of CaCl2, the enzyme retained up to 87-89% activity at temperature range 30°C-40°C and 90-93% activity at temperature range of 50°C-60°C and 89-90% at 70°C.…”

“…Characterization and stabilization of purified enzymes under controlled optimum condition (pH and temperature) are key physiological factors to be considered for maximum enzyme activity and for applications in industries and the environment (Singh et al, 2014). The partially purified peroxidase showed a maximum activity at pH 4.5 and was relatively stable in the pH range between 4.0 and 5.5.…”

Screening and characterization of amylase enzyme in sweet orange (Citrus sinensis) juice clarification

“…Purification fold of the ammonium sulfate precipitation followed by dialysis of crude enzymes ranged from 2.52-10.67. Singh et al (2014) reported that after ammonium sulfate precipitation and dialysis, amylase recovered 56.58% with 2.98-fold purification. Mageswari et al (2012) stated that purification fold of amylase from Pontibacillus chungwhensis strain P1 were 10.1 after ammonium sulfate precipitation and dialysis.…”