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Purification and Characterization of a Polyextremophilic α -Amylase from an Obligate Halophilic Aspergillus penicillioides Isolate and Its Potential for Souse with Detergents.

Ali I, Akbar A, Anwar M, Prasongsuk S, Lotrakul P, Punnapayak H - Biomed Res Int (2015)

Bottom Line: The enzyme was found to have certain polyextremophilic characteristics, with an optimum activity at pH 9, 80 °C, and 300 g · L(-1) NaCl.The addition of CaCl2 at 2 mM was found to slightly enhance the amylase activity, while ZnCl2, FeCl2, or EDTA at 2 mM was strongly or moderately inhibitory, respectively, suggesting the requirement for a (non-Fe(2+) or Zn(2+)) divalent cation.The enzyme retained more than 80% of its activity when incubated with three different laundry detergents and had a better performance compared to a commercial amylase and three detergents in the presence of increasing NaCl concentrations up to 300 g · L(-1).

View Article: PubMed Central - PubMed

Affiliation: Plant Biomass Utilization Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand ; Institute of Biochemistry, University of Balochistan, Quetta 87300, Pakistan.

ABSTRACT
An extracellular α-amylase from the obligate halophilic Aspergillus penicillioides TISTR3639 strain was produced and enriched to apparent homogeneity by ammonium sulfate precipitation and Sephadex G100 gel filtration column chromatography. The mass of the purified amylase was estimated to be 42 kDa by SDS-PAGE. With soluble starch as the substrate it had a specific activity of 118.42 U · mg(-1) and Vmax and Km values of 1.05 µmol · min(-1) · mg(-1) and 5.41 mg · mL(-1), respectively. The enzyme was found to have certain polyextremophilic characteristics, with an optimum activity at pH 9, 80 °C, and 300 g · L(-1) NaCl. The addition of CaCl2 at 2 mM was found to slightly enhance the amylase activity, while ZnCl2, FeCl2, or EDTA at 2 mM was strongly or moderately inhibitory, respectively, suggesting the requirement for a (non-Fe(2+) or Zn(2+)) divalent cation. The enzyme retained more than 80% of its activity when incubated with three different laundry detergents and had a better performance compared to a commercial amylase and three detergents in the presence of increasing NaCl concentrations up to 300 g · L(-1). Accordingly, it has a good potential for use as an α-amylase in a low water activity (high salt concentration) and at high pH and temperatures.

No MeSH data available.


Related in: MedlinePlus

Effect of the (a) pH (at 30°C) and (b) temperature (at pH 9.0) on the activity of the enriched α-amylase from A. penicillioides TISTR 3639. Data are shown as the mean relative activity (%) ±1 SD (error bars), derived from three repeats. Means with a different lowercase superscript letter (a, b, and c) are significantly different (ANOVA and DMRT of the transformed data, P < 0.05).
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fig2: Effect of the (a) pH (at 30°C) and (b) temperature (at pH 9.0) on the activity of the enriched α-amylase from A. penicillioides TISTR 3639. Data are shown as the mean relative activity (%) ±1 SD (error bars), derived from three repeats. Means with a different lowercase superscript letter (a, b, and c) are significantly different (ANOVA and DMRT of the transformed data, P < 0.05).

Mentions: The effect of increasing the pH (range 5–12) on the enriched α-amylase activity, when assayed at 30°C, is shown in Figure 2(a), where the enzyme activity was higher (>80%) in a neutral to moderately alkaline pH (7–10). The enzyme activity declined above and below pH 9, but this decline was more marked with increasing pH above 9 (64% activity at pH 12) than with decreasing pH (~75 and ~73% at pH 6 and 5, resp.).


Purification and Characterization of a Polyextremophilic α -Amylase from an Obligate Halophilic Aspergillus penicillioides Isolate and Its Potential for Souse with Detergents.

Ali I, Akbar A, Anwar M, Prasongsuk S, Lotrakul P, Punnapayak H - Biomed Res Int (2015)

Effect of the (a) pH (at 30°C) and (b) temperature (at pH 9.0) on the activity of the enriched α-amylase from A. penicillioides TISTR 3639. Data are shown as the mean relative activity (%) ±1 SD (error bars), derived from three repeats. Means with a different lowercase superscript letter (a, b, and c) are significantly different (ANOVA and DMRT of the transformed data, P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4477103&req=5

fig2: Effect of the (a) pH (at 30°C) and (b) temperature (at pH 9.0) on the activity of the enriched α-amylase from A. penicillioides TISTR 3639. Data are shown as the mean relative activity (%) ±1 SD (error bars), derived from three repeats. Means with a different lowercase superscript letter (a, b, and c) are significantly different (ANOVA and DMRT of the transformed data, P < 0.05).
Mentions: The effect of increasing the pH (range 5–12) on the enriched α-amylase activity, when assayed at 30°C, is shown in Figure 2(a), where the enzyme activity was higher (>80%) in a neutral to moderately alkaline pH (7–10). The enzyme activity declined above and below pH 9, but this decline was more marked with increasing pH above 9 (64% activity at pH 12) than with decreasing pH (~75 and ~73% at pH 6 and 5, resp.).

Bottom Line: The enzyme was found to have certain polyextremophilic characteristics, with an optimum activity at pH 9, 80 °C, and 300 g · L(-1) NaCl.The addition of CaCl2 at 2 mM was found to slightly enhance the amylase activity, while ZnCl2, FeCl2, or EDTA at 2 mM was strongly or moderately inhibitory, respectively, suggesting the requirement for a (non-Fe(2+) or Zn(2+)) divalent cation.The enzyme retained more than 80% of its activity when incubated with three different laundry detergents and had a better performance compared to a commercial amylase and three detergents in the presence of increasing NaCl concentrations up to 300 g · L(-1).

View Article: PubMed Central - PubMed

Affiliation: Plant Biomass Utilization Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand ; Institute of Biochemistry, University of Balochistan, Quetta 87300, Pakistan.

ABSTRACT
An extracellular α-amylase from the obligate halophilic Aspergillus penicillioides TISTR3639 strain was produced and enriched to apparent homogeneity by ammonium sulfate precipitation and Sephadex G100 gel filtration column chromatography. The mass of the purified amylase was estimated to be 42 kDa by SDS-PAGE. With soluble starch as the substrate it had a specific activity of 118.42 U · mg(-1) and Vmax and Km values of 1.05 µmol · min(-1) · mg(-1) and 5.41 mg · mL(-1), respectively. The enzyme was found to have certain polyextremophilic characteristics, with an optimum activity at pH 9, 80 °C, and 300 g · L(-1) NaCl. The addition of CaCl2 at 2 mM was found to slightly enhance the amylase activity, while ZnCl2, FeCl2, or EDTA at 2 mM was strongly or moderately inhibitory, respectively, suggesting the requirement for a (non-Fe(2+) or Zn(2+)) divalent cation. The enzyme retained more than 80% of its activity when incubated with three different laundry detergents and had a better performance compared to a commercial amylase and three detergents in the presence of increasing NaCl concentrations up to 300 g · L(-1). Accordingly, it has a good potential for use as an α-amylase in a low water activity (high salt concentration) and at high pH and temperatures.

No MeSH data available.


Related in: MedlinePlus