The antioxidant compounds are needed for the food, beverages, and pharmaceuticals industry. For this purpose, an appropriate method is required to measure the antioxidant properties in various types of samples. Spectrophotometric method usually used has some weaknesses, including the high price, long sample preparation time, and less sensitivity. Among the alternative methods developed to overcome these weaknesses is antioxidant biosensor based on superoxide dismutase (SOD) enzyme. Therefore, this study was carried out to measure the SOD activity originating from Deinococcus radiodurans and to determine its kinetics properties. Carbon paste electrode modified with ferrocene and immobilized SOD exhibited anode and cathode current peak at potential of +400 and +300mv respectively, in both pure SOD and SOD of D. radiodurans. This indicated that the current generated was from superoxide catalytic dismutation reaction by SOD. Optimum conditions for SOD activity was at pH 9 and temperature of 27.50C for D. radiodurans SOD, and pH 11 and temperature of 200C for pure SOD. Dismutation reaction kinetics of superoxide catalyzed by SOD followed the Lineweaver-Burk kinetics with D. radiodurans SOD KMapp value was smaller than pure SOD. The result showed that D. radiodurans SOD had higher enzyme-substrate affinity and specificity than pure SOD. It concluded that D. radiodurans SOD had a great potential as biological recognition component for antioxidant biosensor.
<p>[1] S. Chevion, M. A. Roberts, M. Chevion, “The use of cyclic voltammetry
for the evaluation antioxidant capacity,” Free Rad. Biol. Med. vol. 6, no.
28, pp. 860-870, 2000.
[2] R. Dinkov, G. Hristov, D. Stratiev, V. B. Aldayri, “Effect of
commercially available antioxidants over biodiesel/diesel blends
stability,” Fuel. vol. no. 88, pp.732-737, 2009.
[3] L. D. Mello, L.T. Kubota, “Biosensor as a tool for antioxidant status
evaluation,” Talanta, vol. no. 72, pp. 335-348, 2007.
[4] B. Prieto-Simon, M. Cortina, M. Campas, C. Calas-Blanchard,
“Electrochemical biosensor as a tool for antioxidant capacity
assessment,” Sens. Actuators B, vol. no. 129, pp. 459-466, 2008.
[5] L. Campanella, G Favero, L. Persi, M. Tomasetti, “Evaluation of radical
scavenging properties of several plants, fresh or from a herbalist’s using
superoxide dismutase biosensor,” J. Pharm. Biomed. Anal, vol. 24, pp.
1055-1064, 2001.
[6] L. Campanella, A. Bonanni, G. Favero, M. Tomassetti, “Determination
of antioxidant properties of aromatic herbs, olives and fresh fruit using
an enzymatic sensor,” Anal. Bioanal. Chem, vol. 375, pp. 1011-1016,
2003.
[7] L. Campanella, A. Bonanni, M. Tomassetti, “Determination of
antioxidant capacity of samples of different types of tea, or of beverages
based on tea or other herbal product, using a superoxide dismutase
biosensor,” J. Pharm. Biomed. Anal, vol. 32, pp. 725-736, 2003.
[8] L. Campanella, A. Bonanni, E. Finotti, M. Tomassetti, “Biosensors for
determination of total and natural antioxidant capacity of red and white
wines: comparison with other spectrophotometric and fluorimetric
methods,” Biosens. Bielectron, vol. 19, pp. 641-651, 2003.
[9] L. Campanella, E. Martini, M. Tomassetti, “Antioxidant capacity of the
algae using a biosensor method,” Talanta, vol. 66, pp. 902-911, 2005.
[10] L. Campanella, G. Favero, L. Persi, M. Tomasetti, “New biosensor for
superoxide radical used to evidence molecules of biomedical and
pharmaceutical interest having radical scavenging properties,” J. Pharm.
Biomed. Anal, vol. 23, pp. 69-76, 2000.
[11] L. Campanella, A. Bonanni, D. Bellantoni, G. Favero, M. Tomassetti,
“Comparison of fluorimetric, voltammetric and biosensor methods for
the determination of total antioxidant capacity of drug products
containing acetylsalicylic acid,” J. Pharm. Biomed. Anal, vol. 36, pp.
91-99, 2004.
[12] L. Campanella, A. Bonanni, D. Bellantoni, M. Tomassetti, “Biosensor
for determination of total antioxidant capacity of phytotherapeutic
integrators: comparison with other spectrophotometric, fluorimetric
andvoltammetric methods,” J. Pharm. Biomed. Anal, vol. 35, pp. 303-
320, 2004.
[13] L. Campanella, S. De Luca, G. Favero, L. Persi, M. Tomassetti,
“Superoxide dismutase biosensor working in non-aqueous solvent,”
Fresenius J. Anal. Chem, vol. 369, pp. 594-600, 2001.
[14] A. Bonanni, L. Campanella, T. Gatta, E. Gregori, M. Tomassetti,
“Evaluation of the antioxidant and prooxidant properties of several
commercial dry spices by different analytical methods,” Food Chem,vol.
102, pp. 751-758, 2007.
[15] L. T. Benov, W. F. Beyer Jr, R. D. Stevens, I. Fridovich, “Purification
and characterization of the Cu,Zn SOD from Escherichia coli,” Free
Rad Bio Med, vol. 21, no. 1, pp. 117-121, 1996.
[16] M. B. Angelova, S. B. Pashova, L. S. Slokoska, “Comparison of
antioxidant enzyme biosynthesis by free and immobilized Aspergillus
niger cells,” Enzyme Microb. Tech, vol. 26, pp. 544-549, 2000.
[17] E. Zyracka et al., “Ascorbate abolishes auxotrophy caused by the lack of
superoxide dismutase in Saccharomyces cerevisiae Yeast can be a
biosensor for antioxidant,” J Biotech, vol. 115, pp. 271-278, 2005.
[18] R. J. Dennis et al., “Structure of the manganese superoxide dismutase
from Deinococcus radiodurans in two crystal forms,” Acta cryst, vol.
F62, pp. 325-329, 2006.
[19] S. Y. Young, N. L. Young, “Production of superoxide dismutase by
Deinococcus radiophylus,” J. Biochem. Mol. Biol, vol. 36, no. 3, pp.
282-287, 2003.
[20] J. Jia-Ying, S. N. Keeney, E. M. Gregory, “Reconstitution of the
Deinococcus radiodurans aposuperoxide dismutase,” Arch Biochem
Biophysics, vol.286, no. 1, pp. 257-263, 1991.
[21] G. Fang-Xian, E. Shi-Jin, L. Shou-An, C. Jing, L. Duo-Chuan,
“Purification and characterization of a thermostable MnSOD from the
thermophilic fungus Chaetomium thermophilum,” Mycologia, vol. 100,
no. 3, pp. 375-380, 2008.
[22] J. M. McCord, I. Fridovich, “Superoxide dismutase An enzymic for
erythrocuprein (hemocuprein),” J. Biol. Chem, vol. 244, no. 22, pp.
6049-6055, 1969.
[23] L. Campanella, G. Favero, M. Tomasetti, “A modified amperometric
electrode for the determination of free radical,” Sens. Actuators B, vol.
44, pp. 559-565, 1997.
[24] T. Ikeda et al., ”Electrochemical monitoring of in vivo reconstitution of
glucose dehydrogenase in Escherichia coli cells with externally added
pyrroloquinoline quinine,” J. Electroanal. Chem, vol. 449, pp. 219-224,
1998.
[25] A. M. Earl, M. M. Mohundro, I. S. Mian, J. R. Battista, “The irrE
protein of Deinococcus radiodurans R1 is a novel regulator of iA
expression,” J. Bacteriol, vol. 184, no. 22, pp. 6216-6224, 2002.
[26] Y. Zhang et al., “irrE, an exogenous gene from Deinococcus
radiodurans, improves the growth of and ethanol production by a
Zymomonas mobilis strain under ethanol and acid stress,” J. Microbiol.
Biotechnol, vol. 20, no. 7, pp. 1156-1162, 2010.
[27] F. Yamakura, “Purification, crystallization, and properties of ironcontaining
superoxide dismutase from Pseudomonas ovalis,” Biochim.
Biophys. Act, vol. 422, pp. 280-294, 1976.
[28] J.R. Battista, “Against all odds: the survival strategies of Deinococcus
radiodurans,” Annu, Rev. Microbiol, vol. 51, pp. 203-224, 1997.
[29] D. Iswantini, K. Kato, K. Kano, T. Ikeda, “Electrochemical
measurements of glucose dehydrogenase activity exhibited by
Escherichia coli cells; effects of the addition of pyrroloquinoline
quinone, magnesium or calcium ions and ethylenediaminetetraacetic
acid, ”Bioelectrochemistry and Bioenergetics, vol. 46, pp.249-254,
1998.
[30] K. Endo et al., “Development of superoxide sensor by immobilization of
superoxide dismutase,” Sens. Actuators B, vol. 83, pp. 30-34, 2002.
[31] J. Di, S. Peng, C. Shen, Y. Gao, Y. Tu, “One-step method embedding
superoxide dismutase and gold nanoparticles in silica sol-gel network in
the presence of cysteine for construction of third-generation biosensor,”
Biosens. Bioelectron, vol. 23, pp. 88-94, 2007.
[32] R. Antiochia, K. Movassaghi,P Lipone, L. Campanella, “Determination
of the antioxidant capacity of different types of bread and flour using a
superoxide dismutase biosensor, J. Chem. Chem. Eng, vol. 6, pp. 199-
208, 2012.
[33] I. A. Abreu, D. E. Cabelli, “Superoxide dismutases-a review of the
metal-associated mechanistic variations,” BBAPAP, vol. 1804, pp. 263-
274, 2010.
[34] I.A. Abreu et al., “The kinetic mechanism of manganese-containing
superoxide dismutase from Deinococcus radiodurans : A specialized
enzyme for the elimination of high superoxide concentrations,”
Biochem, vol. 47, pp. 2350-2356, 2008.
[35] A. Rigo, P. Viglino, G Rotilio, “Kinetic study of O2
- dismutation by
bovine superoxide dismutase. Evidence for saturation of the catalytic
sites by O2
-,” Biochem Biophysic Res Comm, vol. 63, no. 4, pp. 1013-
1018, 1975.</p>
<p>[1] S. Chevion, M. A. Roberts, M. Chevion, “The use of cyclic voltammetry
for the evaluation antioxidant capacity,” Free Rad. Biol. Med. vol. 6, no.
28, pp. 860-870, 2000.
[2] R. Dinkov, G. Hristov, D. Stratiev, V. B. Aldayri, “Effect of
commercially available antioxidants over biodiesel/diesel blends
stability,” Fuel. vol. no. 88, pp.732-737, 2009.
[3] L. D. Mello, L.T. Kubota, “Biosensor as a tool for antioxidant status
evaluation,” Talanta, vol. no. 72, pp. 335-348, 2007.
[4] B. Prieto-Simon, M. Cortina, M. Campas, C. Calas-Blanchard,
“Electrochemical biosensor as a tool for antioxidant capacity
assessment,” Sens. Actuators B, vol. no. 129, pp. 459-466, 2008.
[5] L. Campanella, G Favero, L. Persi, M. Tomasetti, “Evaluation of radical
scavenging properties of several plants, fresh or from a herbalist’s using
superoxide dismutase biosensor,” J. Pharm. Biomed. Anal, vol. 24, pp.
1055-1064, 2001.
[6] L. Campanella, A. Bonanni, G. Favero, M. Tomassetti, “Determination
of antioxidant properties of aromatic herbs, olives and fresh fruit using
an enzymatic sensor,” Anal. Bioanal. Chem, vol. 375, pp. 1011-1016,
2003.
[7] L. Campanella, A. Bonanni, M. Tomassetti, “Determination of
antioxidant capacity of samples of different types of tea, or of beverages
based on tea or other herbal product, using a superoxide dismutase
biosensor,” J. Pharm. Biomed. Anal, vol. 32, pp. 725-736, 2003.
[8] L. Campanella, A. Bonanni, E. Finotti, M. Tomassetti, “Biosensors for
determination of total and natural antioxidant capacity of red and white
wines: comparison with other spectrophotometric and fluorimetric
methods,” Biosens. Bielectron, vol. 19, pp. 641-651, 2003.
[9] L. Campanella, E. Martini, M. Tomassetti, “Antioxidant capacity of the
algae using a biosensor method,” Talanta, vol. 66, pp. 902-911, 2005.
[10] L. Campanella, G. Favero, L. Persi, M. Tomasetti, “New biosensor for
superoxide radical used to evidence molecules of biomedical and
pharmaceutical interest having radical scavenging properties,” J. Pharm.
Biomed. Anal, vol. 23, pp. 69-76, 2000.
[11] L. Campanella, A. Bonanni, D. Bellantoni, G. Favero, M. Tomassetti,
“Comparison of fluorimetric, voltammetric and biosensor methods for
the determination of total antioxidant capacity of drug products
containing acetylsalicylic acid,” J. Pharm. Biomed. Anal, vol. 36, pp.
91-99, 2004.
[12] L. Campanella, A. Bonanni, D. Bellantoni, M. Tomassetti, “Biosensor
for determination of total antioxidant capacity of phytotherapeutic
integrators: comparison with other spectrophotometric, fluorimetric
andvoltammetric methods,” J. Pharm. Biomed. Anal, vol. 35, pp. 303-
320, 2004.
[13] L. Campanella, S. De Luca, G. Favero, L. Persi, M. Tomassetti,
“Superoxide dismutase biosensor working in non-aqueous solvent,”
Fresenius J. Anal. Chem, vol. 369, pp. 594-600, 2001.
[14] A. Bonanni, L. Campanella, T. Gatta, E. Gregori, M. Tomassetti,
“Evaluation of the antioxidant and prooxidant properties of several
commercial dry spices by different analytical methods,” Food Chem,vol.
102, pp. 751-758, 2007.
[15] L. T. Benov, W. F. Beyer Jr, R. D. Stevens, I. Fridovich, “Purification
and characterization of the Cu,Zn SOD from Escherichia coli,” Free
Rad Bio Med, vol. 21, no. 1, pp. 117-121, 1996.
[16] M. B. Angelova, S. B. Pashova, L. S. Slokoska, “Comparison of
antioxidant enzyme biosynthesis by free and immobilized Aspergillus
niger cells,” Enzyme Microb. Tech, vol. 26, pp. 544-549, 2000.
[17] E. Zyracka et al., “Ascorbate abolishes auxotrophy caused by the lack of
superoxide dismutase in Saccharomyces cerevisiae Yeast can be a
biosensor for antioxidant,” J Biotech, vol. 115, pp. 271-278, 2005.
[18] R. J. Dennis et al., “Structure of the manganese superoxide dismutase
from Deinococcus radiodurans in two crystal forms,” Acta cryst, vol.
F62, pp. 325-329, 2006.
[19] S. Y. Young, N. L. Young, “Production of superoxide dismutase by
Deinococcus radiophylus,” J. Biochem. Mol. Biol, vol. 36, no. 3, pp.
282-287, 2003.
[20] J. Jia-Ying, S. N. Keeney, E. M. Gregory, “Reconstitution of the
Deinococcus radiodurans aposuperoxide dismutase,” Arch Biochem
Biophysics, vol.286, no. 1, pp. 257-263, 1991.
[21] G. Fang-Xian, E. Shi-Jin, L. Shou-An, C. Jing, L. Duo-Chuan,
“Purification and characterization of a thermostable MnSOD from the
thermophilic fungus Chaetomium thermophilum,” Mycologia, vol. 100,
no. 3, pp. 375-380, 2008.
[22] J. M. McCord, I. Fridovich, “Superoxide dismutase An enzymic for
erythrocuprein (hemocuprein),” J. Biol. Chem, vol. 244, no. 22, pp.
6049-6055, 1969.
[23] L. Campanella, G. Favero, M. Tomasetti, “A modified amperometric
electrode for the determination of free radical,” Sens. Actuators B, vol.
44, pp. 559-565, 1997.
[24] T. Ikeda et al., ”Electrochemical monitoring of in vivo reconstitution of
glucose dehydrogenase in Escherichia coli cells with externally added
pyrroloquinoline quinine,” J. Electroanal. Chem, vol. 449, pp. 219-224,
1998.
[25] A. M. Earl, M. M. Mohundro, I. S. Mian, J. R. Battista, “The irrE
protein of Deinococcus radiodurans R1 is a novel regulator of iA
expression,” J. Bacteriol, vol. 184, no. 22, pp. 6216-6224, 2002.
[26] Y. Zhang et al., “irrE, an exogenous gene from Deinococcus
radiodurans, improves the growth of and ethanol production by a
Zymomonas mobilis strain under ethanol and acid stress,” J. Microbiol.
Biotechnol, vol. 20, no. 7, pp. 1156-1162, 2010.
[27] F. Yamakura, “Purification, crystallization, and properties of ironcontaining
superoxide dismutase from Pseudomonas ovalis,” Biochim.
Biophys. Act, vol. 422, pp. 280-294, 1976.
[28] J.R. Battista, “Against all odds: the survival strategies of Deinococcus
radiodurans,” Annu, Rev. Microbiol, vol. 51, pp. 203-224, 1997.
[29] D. Iswantini, K. Kato, K. Kano, T. Ikeda, “Electrochemical
measurements of glucose dehydrogenase activity exhibited by
Escherichia coli cells; effects of the addition of pyrroloquinoline
quinone, magnesium or calcium ions and ethylenediaminetetraacetic
acid, ”Bioelectrochemistry and Bioenergetics, vol. 46, pp.249-254,
1998.
[30] K. Endo et al., “Development of superoxide sensor by immobilization of
superoxide dismutase,” Sens. Actuators B, vol. 83, pp. 30-34, 2002.
[31] J. Di, S. Peng, C. Shen, Y. Gao, Y. Tu, “One-step method embedding
superoxide dismutase and gold nanoparticles in silica sol-gel network in
the presence of cysteine for construction of third-generation biosensor,”
Biosens. Bioelectron, vol. 23, pp. 88-94, 2007.
[32] R. Antiochia, K. Movassaghi,P Lipone, L. Campanella, “Determination
of the antioxidant capacity of different types of bread and flour using a
superoxide dismutase biosensor, J. Chem. Chem. Eng, vol. 6, pp. 199-
208, 2012.
[33] I. A. Abreu, D. E. Cabelli, “Superoxide dismutases-a review of the
metal-associated mechanistic variations,” BBAPAP, vol. 1804, pp. 263-
274, 2010.
[34] I.A. Abreu et al., “The kinetic mechanism of manganese-containing
superoxide dismutase from Deinococcus radiodurans : A specialized
enzyme for the elimination of high superoxide concentrations,”
Biochem, vol. 47, pp. 2350-2356, 2008.
[35] A. Rigo, P. Viglino, G Rotilio, “Kinetic study of O2
- dismutation by
bovine superoxide dismutase. Evidence for saturation of the catalytic
sites by O2
-,” Biochem Biophysic Res Comm, vol. 63, no. 4, pp. 1013-
1018, 1975.</p>
@article{"International Journal of Medical, Medicine and Health Sciences:60246", author = "Dyah Iswantini and Trivadila and Novik Nurhidayat and Waras Nurcholis", title = "Antioxidant Biosensor Using Microbe", abstract = "The antioxidant compounds are needed for the food, beverages, and pharmaceuticals industry. For this purpose, an appropriate method is required to measure the antioxidant properties in various types of samples. Spectrophotometric method usually used has some weaknesses, including the high price, long sample preparation time, and less sensitivity. Among the alternative methods developed to overcome these weaknesses is antioxidant biosensor based on superoxide dismutase (SOD) enzyme. Therefore, this study was carried out to measure the SOD activity originating from Deinococcus radiodurans and to determine its kinetics properties. Carbon paste electrode modified with ferrocene and immobilized SOD exhibited anode and cathode current peak at potential of +400 and +300mv respectively, in both pure SOD and SOD of D. radiodurans. This indicated that the current generated was from superoxide catalytic dismutation reaction by SOD. Optimum conditions for SOD activity was at pH 9 and temperature of 27.50C for D. radiodurans SOD, and pH 11 and temperature of 200C for pure SOD. Dismutation reaction kinetics of superoxide catalyzed by SOD followed the Lineweaver-Burk kinetics with D. radiodurans SOD KMapp value was smaller than pure SOD. The result showed that D. radiodurans SOD had higher enzyme-substrate affinity and specificity than pure SOD. It concluded that D. radiodurans SOD had a great potential as biological recognition component for antioxidant biosensor.
", keywords = "Antioxidant biosensor, Deinococcus radiodurans,
enzyme kinetic, superoxide dismutase (SOD).", volume = "7", number = "6", pages = "289-8", }
