Pectin methylesterase (PME) extracted from muskmelon was purified by anion exchange chromatography. The specific activity of purified enzyme was 152.01 U/mg and relative molecular weight was ~69,000 Da. Methylesterase was characterized for various physicochemical factors to designate its suitability in the food industry applications. The optimum temperature of the enzyme was 30°C and is thermally stable between the temperature ranges of 15-65°C with critical temperature for stability being >65°C. Thermal inactivation first order kinetics and thermodynamic parameters in temperature range (45-65°C) favors stability of PME and at 75°C complete inactivation of enzyme was observed indicating the unstable nature of enzyme over >65°C. Activation energy (Ea ) and Z values of thermal inactivation were found to be 100.108 kJ/mol and 2.05°C, respectively. About 0.1 M NaCl is essential for enzyme to attain the maximum activity. The enzyme lost activity in presence of divalent calcium (Ca2+ ) and magnesium (Mg2+ ) ions. PRACTICAL APPLICATIONS: Pectin methylesterase (EC3.1.1.11) are an important class of enzymes expressed in plants and microbes and they bring about the de-methylesterification on pectin substrate. Up to ~13% degree of esterification of pectin was observed with muskmelon PME enzyme treatment. The de-methylesterified pectin thus prepared was subjected for gelation in presence of Ca2+ ions and above 0.5% of demethylesterified pectin stable calcium pectate gels were produced. The study demonstrates the suitability of muskmelon PME extracted from biowaste in food applications with good gelling property.