Production and characterization of cellobiose dehydrogenase from Phanerochaete chrysosporium KCCM 60256 and its application for an enzymatic fuel cell

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• 作者：Han Suk Choi ; Dong Sup Kim ; Laxmi Prasad Thapa…
• 关键词：Cellobiose Dehydrogenase ; Pichia pastoris ; Phanerochaete chrysosporium ; Enzymatic Fuel Cell
• 刊名：Korean Journal of Chemical Engineering
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：33
• 期：12
• 页码：3434-3441
• 全文大小：
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Industrial Chemistry/Chemical Engineering; Catalysis; Materials Science, general; Biotechnology;
• 出版者：Springer US
• ISSN：1975-7220
• 卷排序：33

文摘

The enzyme cellobiose dehydrogenase (CDH), with high ability of electron transport, has been widely used in enzymatic fuel cells or biosensors. In this study, the cellobiose dehydrogenase gene from Phanerochaete chrysosporium KCCM 60256 was amplified and expressed in the methylotrophic yeast Pichia pastoris X-33. The recombinant enzyme (PcCDH) was purified using a metal affinity chromatography under non-denaturing conditions. The purified enzyme was analyzed by SDS-PAGE, confirming a corresponding band about 100 kDa. The enzyme activity of this purified PcCDH was determined as 1,845U/L (65mg/L protein). The enzyme showed the maximum activity at pH 4.5 and high activity in broad ranges of temperature from 30°C to 60°C. Moreover, the application of PcCDH to enzymatic fuel cell (EFC) was demonstrated. Lactose was used as the substrate in the EFC system; anode and cathode were immobilized with PcCDH and laccase, respectively. The cell’s open circuit voltage and maximum power density of the EFC system were, respectively, determined as 0.435 V and 314 μW/cm2 (at 0.247 V) with 10 mM lactose.