Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 1-8 |
| Number of pages | 8 |
| Journal | Korean Journal of Chemical Engineering |
| DOIs | |
| Publication status | Accepted/In press - 2016 Sep 3 |
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Keywords
- Cellobiose Dehydrogenase
- Enzymatic Fuel Cell
- Phanerochaete chrysosporium
- Pichia pastoris
ASJC Scopus subject areas
- Chemistry(all)
- Chemical Engineering(all)
Cite this
Production and characterization of cellobiose dehydrogenase from Phanerochaete chrysosporium KCCM 60256 and its application for an enzymatic fuel cell. / Choi, Han Suk; Kim, Dong Sup; Thapa, Laxmi Prasad; Lee, Sang Jun; Kim, Sung Bong; Cho, Jaehoon; Park, Chulhwan; Kim, Seung Wook.
In: Korean Journal of Chemical Engineering, 03.09.2016, p. 1-8.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Production and characterization of cellobiose dehydrogenase from Phanerochaete chrysosporium KCCM 60256 and its application for an enzymatic fuel cell
AU - Choi, Han Suk
AU - Kim, Dong Sup
AU - Thapa, Laxmi Prasad
AU - Lee, Sang Jun
AU - Kim, Sung Bong
AU - Cho, Jaehoon
AU - Park, Chulhwan
AU - Kim, Seung Wook
PY - 2016/9/3
Y1 - 2016/9/3
N2 - 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.
AB - 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.
KW - Cellobiose Dehydrogenase
KW - Enzymatic Fuel Cell
KW - Phanerochaete chrysosporium
KW - Pichia pastoris
UR - http://www.scopus.com/inward/record.url?scp=84984911590&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84984911590&partnerID=8YFLogxK
U2 - 10.1007/s11814-016-0205-4
DO - 10.1007/s11814-016-0205-4
M3 - Article
AN - SCOPUS:84984911590
SP - 1
EP - 8
JO - Korean Journal of Chemical Engineering
JF - Korean Journal of Chemical Engineering
SN - 0256-1115
ER -