Kinetics and Thermodynamic Properties of Pectinase Obtained from Trichoderma longibrachiatum MT321074
. http://www.doi.org/10.26538/tjnpr/v6i12.28
Abstract
Thermo-instability affects enzymes' efficiency in the industry. This study explored the
thermodynamic and kinetic properties of Trichoderma longibrachiatum MT321074 pectinase, to
ascertain its potential for industrial application. Trichoderma longibrachiatum MT321074 was
isolated from decaying mango fruits and identified using the 18S rDNA sequencing technique.
The Trichoderma longibrachiatum MT321074 pectinase was purified 5.61 fold with a specific
activity and yield of 2.86 U/mg and 17.63%. The pH and temperature optima were 5.0 and 40oC.
The Michaelis constant, KM, and maximum velocity, Vmax were 1.0 mg/ml and 82.64 µmol/min.
The enzyme retained 92%, 87%, and 72% of its original activity after 2 h at 30ºC, 40ºC, and
50ºC, respectively. At 60ºC, 70ºC and 80ºC, the enzyme lost 38%, 65%, and 73% of its activity
after 120 min. The enzyme's melting temperature (Tm) was 64oC. The rates of thermal
inactivation (Kd) were low at all temperatures tested. The activation energy of inactivation (Ea)
was 50.30 KJ/mol-1. The D-values and half-life (t1/2) decreased from 3838.33 to 261.70 min and
1155.00 to 78.75 min, respectively. The enzyme had a high Z-value of 42oC. The Gibbs free
energy (ΔGin) increased with a rise in temperature. High enthalpy changes (ΔHin) were obtained
at all temperatures tested. The entropy (ΔSin) values were negative at all temperatures tested. In
summary, the increase in ∆Gin accompanied by decline in ΔSin and ∆Hin values at elevated
temperatures suggests that the enzyme is heat stable and thus, very suitable for the degradation
of pectic substances in fruit effluents, food, and beverage products.
Keywords: Trichoderma longibrachiatum MT321074, 18S-rDNA sequencing, Pectinase, Heat
stability
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