Production of a novel pectin lyase from Bacillus pumilus (P9), Purification and characterisation and fruit juice application

Abstract

Extracellular pectin lyase (EC 4.2.2.10) was produced by Bacillus pumilus (P9) in solid state fermentation. Pectin lyase enzyme was purified 36.36 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 25 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was controlled by SDS-PAGE. The optimum pH and temperature of enzyme was determined as pH 6.0 and 60oC, respectively. Pectin lyase was mostly stable at 40°C. Its' activity deceased in 50% for 1h at 60°C and 40% for 4 h at 50°C. Vmax and KM were calculated respectively as 0.298 mg/mL and 132.6 mol/L*min. The presence of 10 mM concentration of Ca2+, Cu2+, Mn2+, Mg2+, Zn2+, Hg2+, Fe2+ and EDTA, L-cystein, ascorbic acid and -mercaptoethanol significantly enhanced the pectin lyase of the purified enzyme. The purified pectin lyase enzyme was used for getting fruits juices. It was determined that yields of fruits juices significantly improved when it was compared with control. © 2010 University of Bucharest.