Fabrication of functional biomolecular layer using recombinant technique for the bioelectronic device

Sang-Uk Kim; Young Jun Kim; Cheol-Heon Yea; Junhong Min; Jeong-Woo Choi

Korean Journal of Chemical Engineering, Vol.25, No.5, 1115-1119, September, 2008

DOI10.1007/s11814-008-0182-3 Export Citation

Fabrication of functional biomolecular layer using recombinant technique for the bioelectronic device

Sang-Uk Kim¹, Young Jun Kim¹, Cheol-Heon Yea², Junhong Min³, and Jeong-Woo Choi^{1, 2, †}

¹Interdisciplinary Program of Integrated Biotechnology, Sogang University, Seoul 121-742, Korea
²Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Korea
³Department of BioNano technology, Kyungwon University, Seongnam 461-701, Korea

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A novel immobilization method of blue copper protein azurin on a gold surface was developed without a chemical linker using recombinant technique. Azurin was recombined with a cysteine anchors by site-directed mutagenesis (SDM) and used to effect the mutations, changing the codon for Leu39Cys (L39C) from CTG to TGC. The immobilization of the functionalized protein is confirmed by surface plasmon resonance (SPR) and its surface morphology is analyzed by scanning tunneling microscopy (STM). The immobilization efficiency has been increased about 76.3%, as compared to that of wild type azurin. The electrochemical property of the fabricated thin film was investigated by cyclic voltammetry (CV). As a result, cysteine-modified azurin can be used for making high quality protein film, and applied to the fabrication of nano-scale bioelectronics.

Keywords:Bioelectronic;Recombinant Azurin;Cyclic Voltammetry;Site-directed Mutagenesis

[References]

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Baron R, Lioubashevski O, Katz E, Niazov T, Willner I, J. Phys. Chem. A, 110(27), 8548 (2006)
Choi JW, Nam YS, Park SJ, Lee WH, Kim DH, Fujihira M, Biosens. Bioelectron., 16, 819 (2001)
Nam YS, Choi JW, Lee WH, Appl. Phys. Lett., 85, 6275 (2004)
Choi JW, Fujihira M, Appl. Phys. Lett., 84, 2187 (2004)
Choi JW, Nam YS, Lee WH, Appl. Phys. Lett., 79, 1570 (2001)
Porter MD, Bright TB, Allara DL, Chidsey CED, J. Am. Chem. Soc., 39, 11808 (2000)
Dubrovsky TB, Hou Z, Stroeve P, Abbott NL, Anal. Chem., 71, 327 (1999)
Sambrook J, Fritsch EF, Maniatis M, Molecular cloning, Cold Spring Harbor Laboratory Press, New York (2001)
Son YJ, Park KH, Lee SY, Oh SJ, Kim CK, Choi BT, Park YC, Seo JH, Biotechnol. Bioprocess Eng., 12, 556 (2007)
Kim SH, Shin BH, Kim YH, Nam SW, Jeon SJ, Biotechnol. Bioprocess Eng., 12, 707 (2007)
Nichols JD, Xiang S, Schindelin H, Rajagopalan KV, Biochemistry, 46, 78 (2007)
Foley TL, Burkart MD, Curr. Opin. Chem. Biol., 11, 12 (2007)
Wittung-Stafshede P, Inorg. Chem., 43(25), 7926 (2004)
Cavalleri O, Natale C, Stroppolo ME, Relini A, Cosulich E, Thea S, Novie M, Gliozzia A, Phys. Chem. Chem. Phys., 2, 4630 (2000)
Patel N, Davies MC, Hartshorne M, Heaton RJ, Roberts CJ, Tendler SJ, Williams PM, Langmuir, 13(24), 6485 (1997)
Jordan CE, Corn RM, Anal. Chem., 69, 1449 (1997)
Iwatani S, Iwane AH, Higuchi H, Ishii Y, Yanagida T, Biochemistry, 38, 10318 (1999)
Choi JW, Kim JS, Lee BH, Jang YH, Kim YJ, Mol. Cryst. Liq. Cryst., 463, 281 (2007)
Bott AW, Current Separations, 18, 2 (1999)

[Related Articles]

[1] Willner I, Willner B, J. Mater. Chem., 8, 2543 (1998)

[2] Ballardini B, Balzani V, Credi A, Gandolfi MT, Venturi M, Accounts Chem. Res., 34, 445 (2001)

[3] Raymo FM, Adv. Mater., 14(6), 401 (2002)

[4] Hopfield JJ, Onuchic JN, Beratan DN, J. Phys. Chem., 93, 6350 (1989)

[5] Guisinger NP, Yoder NL, Hersam MC, Proc. Natl. Acad. Sci. U.S.A., 102, 8838 (2005)

[6] Roth KM, Lindsey JS, Bocian DF, Kuhr WG, Langmuir, 18(10), 4030 (2002)

[7] Liu Z, Yasseri AA, Lindsey JS, Bocian DF, Science, 302, 1543 (2003)

[8] Weizmann Y, Elnathan R, Lioubashevski O, Willner I, J. Am. Chem. Soc., 127(36), 12666 (2005)

[9] Baron R, Lioubashevski O, Katz E, Niazov T, Willner I, J. Phys. Chem. A, 110(27), 8548 (2006)

[10] Choi JW, Nam YS, Park SJ, Lee WH, Kim DH, Fujihira M, Biosens. Bioelectron., 16, 819 (2001)

[11] Nam YS, Choi JW, Lee WH, Appl. Phys. Lett., 85, 6275 (2004)

[12] Choi JW, Fujihira M, Appl. Phys. Lett., 84, 2187 (2004)

[13] Choi JW, Nam YS, Lee WH, Appl. Phys. Lett., 79, 1570 (2001)

[14] Porter MD, Bright TB, Allara DL, Chidsey CED, J. Am. Chem. Soc., 39, 11808 (2000)

[15] Dubrovsky TB, Hou Z, Stroeve P, Abbott NL, Anal. Chem., 71, 327 (1999)

[16] Sambrook J, Fritsch EF, Maniatis M, Molecular cloning, Cold Spring Harbor Laboratory Press, New York (2001)

[17] Son YJ, Park KH, Lee SY, Oh SJ, Kim CK, Choi BT, Park YC, Seo JH, Biotechnol. Bioprocess Eng., 12, 556 (2007)

[18] Kim SH, Shin BH, Kim YH, Nam SW, Jeon SJ, Biotechnol. Bioprocess Eng., 12, 707 (2007)

[19] Nichols JD, Xiang S, Schindelin H, Rajagopalan KV, Biochemistry, 46, 78 (2007)

[20] Foley TL, Burkart MD, Curr. Opin. Chem. Biol., 11, 12 (2007)

[21] Wittung-Stafshede P, Inorg. Chem., 43(25), 7926 (2004)

[22] Cavalleri O, Natale C, Stroppolo ME, Relini A, Cosulich E, Thea S, Novie M, Gliozzia A, Phys. Chem. Chem. Phys., 2, 4630 (2000)

[23] Patel N, Davies MC, Hartshorne M, Heaton RJ, Roberts CJ, Tendler SJ, Williams PM, Langmuir, 13(24), 6485 (1997)

[24] Jordan CE, Corn RM, Anal. Chem., 69, 1449 (1997)

[25] Iwatani S, Iwane AH, Higuchi H, Ishii Y, Yanagida T, Biochemistry, 38, 10318 (1999)

[26] Choi JW, Kim JS, Lee BH, Jang YH, Kim YJ, Mol. Cryst. Liq. Cryst., 463, 281 (2007)

[27] Bott AW, Current Separations, 18, 2 (1999)