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

Author:
Lu, Xiaohua, Lin, Jianguo, Wang, Changgao, Du, Xin, Cai, Jun
Source:
Food science and biotechnology 2016 v.25 no.5 pp. 1379-1385
ISSN:
1226-7708
Subject:
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
Abstract:
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%.
Agid:
5580132