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Extracellular α-Galactosidase from Trichoderma sp. (WF-3): Optimization of Enzyme Production and Biochemical Characterization.

Chauhan AS, Kumar A, Siddiqi NJ, Sharma B - Biotechnol Res Int (2015)

Bottom Line: Trichoderma spp. have been reported earlier for their excellent capacity of secreting extracellular α-galactosidase.It followed Michaelis-Menten curve and showed direct relationship with varying substrate concentrations.This information would be helpful in understanding the biophysical and biochemical characteristics of extracellular α-galactosidase from other microbial sources.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Science, University of Allahabad, Allahabad 211002, India.

ABSTRACT
Trichoderma spp. have been reported earlier for their excellent capacity of secreting extracellular α-galactosidase. This communication focuses on the optimization of culture conditions for optimal production of enzyme and its characterization. The evaluation of the effects of different enzyme assay parameters such as stability, pH, temperature, substrate concentrations, and incubation time on enzyme activity has been made. The most suitable buffer for enzyme assay was found to be citrate phosphate buffer (50 mM, pH 6.0) for optimal enzyme activity. This enzyme was fairly stable at higher temperature as it exhibited 72% activity at 60°C. The enzyme when incubated at room temperature up to two hours did not show any significant loss in activity. It followed Michaelis-Menten curve and showed direct relationship with varying substrate concentrations. Higher substrate concentration was not inhibitory to enzyme activity. The apparent Michaelis-Menten constant (K m ), maximum rate of reaction (V max), K cat, and catalytic efficiency values for this enzyme were calculated from the Lineweaver-Burk double reciprocal plot and were found to be 0.5 mM, 10 mM/s, 1.30 U mg(-1), and 2.33 U mg(-1) mM(-1), respectively. This information would be helpful in understanding the biophysical and biochemical characteristics of extracellular α-galactosidase from other microbial sources.

No MeSH data available.


Related in: MedlinePlus

Effect of temperature on enzyme activity. The enzyme was assayed at different temperatures as described in Section 2.
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fig1: Effect of temperature on enzyme activity. The enzyme was assayed at different temperatures as described in Section 2.

Mentions: The enzyme activities were assayed under the standard assay conditions at pH 5 for 10 min at various temperatures ranging from 30 to 80°C. Figure 1 shows the temperature-activity profile of α-galactosidase. The enzyme showed maximum activity at 60°C.


Extracellular α-Galactosidase from Trichoderma sp. (WF-3): Optimization of Enzyme Production and Biochemical Characterization.

Chauhan AS, Kumar A, Siddiqi NJ, Sharma B - Biotechnol Res Int (2015)

Effect of temperature on enzyme activity. The enzyme was assayed at different temperatures as described in Section 2.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4644822&req=5

fig1: Effect of temperature on enzyme activity. The enzyme was assayed at different temperatures as described in Section 2.
Mentions: The enzyme activities were assayed under the standard assay conditions at pH 5 for 10 min at various temperatures ranging from 30 to 80°C. Figure 1 shows the temperature-activity profile of α-galactosidase. The enzyme showed maximum activity at 60°C.

Bottom Line: Trichoderma spp. have been reported earlier for their excellent capacity of secreting extracellular α-galactosidase.It followed Michaelis-Menten curve and showed direct relationship with varying substrate concentrations.This information would be helpful in understanding the biophysical and biochemical characteristics of extracellular α-galactosidase from other microbial sources.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Faculty of Science, University of Allahabad, Allahabad 211002, India.

ABSTRACT
Trichoderma spp. have been reported earlier for their excellent capacity of secreting extracellular α-galactosidase. This communication focuses on the optimization of culture conditions for optimal production of enzyme and its characterization. The evaluation of the effects of different enzyme assay parameters such as stability, pH, temperature, substrate concentrations, and incubation time on enzyme activity has been made. The most suitable buffer for enzyme assay was found to be citrate phosphate buffer (50 mM, pH 6.0) for optimal enzyme activity. This enzyme was fairly stable at higher temperature as it exhibited 72% activity at 60°C. The enzyme when incubated at room temperature up to two hours did not show any significant loss in activity. It followed Michaelis-Menten curve and showed direct relationship with varying substrate concentrations. Higher substrate concentration was not inhibitory to enzyme activity. The apparent Michaelis-Menten constant (K m ), maximum rate of reaction (V max), K cat, and catalytic efficiency values for this enzyme were calculated from the Lineweaver-Burk double reciprocal plot and were found to be 0.5 mM, 10 mM/s, 1.30 U mg(-1), and 2.33 U mg(-1) mM(-1), respectively. This information would be helpful in understanding the biophysical and biochemical characteristics of extracellular α-galactosidase from other microbial sources.

No MeSH data available.


Related in: MedlinePlus