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KMID : 1007520160250051379
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Food Science and Biotechnology
2016 Volume.25 No. 5 p.1379 ~ p.1385
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Purification and characterization of exo-polygalacturonase from Zygoascus hellenicus V25 and its potential application in fruit juice clarification
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Lu Xiao Hua
Lin Jian Guo
Wang Chang Gao
Du Xin
Cai Jun
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Abstract
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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%.
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KEYWORD
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Zygoascus hellenicus V25, exo-polygalacturonase, purification, characterization, juice clarification
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