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http://dx.doi.org/10.4014/jmb.1511.11045

A Novel Acid-Stable Endo-Polygalacturonase from Penicillium oxalicum CZ1028: Purification, Characterization, and Application in the Beverage Industry  

Cheng, Zhong (State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology, Guangxi University)
Chen, Dong (National Engineering Research Center for Non-Food Biorefinery, Guangxi Academy of Sciences)
Lu, Bo (National Engineering Research Center for Non-Food Biorefinery, Guangxi Academy of Sciences)
Wei, Yutuo (State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology, Guangxi University)
Xian, Liang (State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology, Guangxi University)
Li, Yi (National Engineering Research Center for Non-Food Biorefinery, Guangxi Academy of Sciences)
Luo, Zhenzhen (Guangxi Zhuang Autonomous Region Institute of Supervision and Testing on Product Quality)
Huang, Ribo (State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Life Science and Technology, Guangxi University)
Publication Information
Journal of Microbiology and Biotechnology / v.26, no.6, 2016 , pp. 989-998 More about this Journal
Abstract
Acidic endo-polygalacturonases are the major part of pectinase preparations and extensively applied in the clarification of fruits juice, vegetables extracts, and wines. However, most of the reported fungal endo-polygalacturonases are active and stable under narrow pH range and low temperatures. In this study, an acidic endo-polygalacturonase (EPG4) was purified and characterized from a mutant strain of Penicillium oxalicum. The N-terminal amino acid sequence of EPG4 (ATTCTFSGSNGAASASKSQT) was different from those of reported endo-polygalacturonases. EPG4 displayed optimal pH and temperature at 5.0 and 60-70℃ towards polygalacturonic acid (PGA), respectively, and was notably stable at pH 2.2-7.0. When tested against pectins, EPG4 showed enzyme activity over a broad acidic pH range (>15.0% activity at pH 2.2-6.0 towards citrus pectin; and >26.6% activity at pH 2.2-7.0 towards apple pectin). The Km and Vmax values were determined as 1.27 mg/ml and 5,504.6 U/mg, respectively. The enzyme hydrolyzed PGA in endo-manner, releasing oligo-galacturonates from PGA, as determined by TLC. Addition of EPG4 (3.6 U/ml) significantly reduced the viscosity (by 42.4%) and increased the light transmittance (by 29.5%) of the papaya pulp, and increased the recovery (by 24.4%) of the papaya extraction. All of these properties make the enzyme a potential application in the beverage industry.
Keywords
Penicillium oxalicum; endo-polygalacturonase; purification; acid stablity; application;
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