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Purification and characterization of exo-polygalacturonase from Zygoascus hellenicus V25 and its potential application in fruit juice clarification

Lu, Xiaohua, Lin, Jianguo, Wang, Changgao, Du, Xin, Cai, Jun
Food science and biotechnology 2016 v.25 no.5 pp. 1379-1385
Zygoascus, ammonium sulfate, cellulose, clarification (processing), copper, correlation, cysteine, enzymatic hydrolysis, enzyme activity, esterification, fruit juices, fruits, gel chromatography, mercury, molecular weight, orange peels, pH, pectins, polygalacturonase, transmittance, wastes
The purification and characterization of the extracellular polygalacturonase from Zygoascus hellenicus V25 submerged culture using orange peel waste were investigated. This polygalacturonase, with a molecular weight of 75.28 kDa, was purified to 16.89 purification fold with a recovery of 18.46% and specific activity of 2469.77 U/mg protein by ammonium sulfate precipitation, DEAE cellulose chromatography, and Sephadex G-100 gel filtration. The enzyme exhibited maximum activity at 60°C and pH 5.0 and was stable over a wide range of pH levels (3.0-11.0). Moreover, enzyme activity was enhanced by Cu²⁺ and cysteine, whereas it was strongly inhibited by Hg²⁺. The extent of enzymatic hydrolysis was negatively correlated with the degree of pectin esterification. Kₘ and Vₘₐₓ values of the polygalacturonase were 5.44 mg/mL and 61.73 μmol/(min·mg), respectively. The polygalacturonase was applied in the juice clarification of four fruits, and results showed that the percentage transmittance at 660 nm increased by 3.51, 4.36, 8.04, and 12.2%.