Share This Article:

Gold Nanoparticles for Colorimetric detection of hydrolysis of antibiotics by penicillin G acylase

Abstract Full-Text HTML XML Download Download as PDF (Size:1762KB) PP. 322-329
DOI: 10.4236/abb.2010.14042    8,018 Downloads   19,568 Views   Citations

ABSTRACT

A simple inexpensive method of monitoring hydrolysis of an antibiotic penicillin G (pen G) and subsequent enzyme detection using gold nanoparticles is presented. Gold nanoparticles capped with Cetyl trimethyl ammonium bromide (CTAB) are synthesized using chemical route. The particles could be used for detection of Penicillin G acylase (PGA) enzyme by incorporating hydrolysis reaction with pen G. This hydrolysis reaction leads to a shift in the surface plasmon band of gold nanoparticles from 527 nm to 545 nm accompanied by a visual colorimetric change in the solution from red to blue. The process is attributed to aggregation of nanoparticles caused due to displacement of CTAB bilayer by byproducts of the hydrolysis reaction. It is proposed that the presence of 0.007 mg/ml of PGA can be detected by a color change of gold nanoparticles solution without requiring any complicated instrument or highly trained operator to conduct the test. The method could also identify the presence of different penicillins by showing different spectral shifts. Thus the work presented here would be useful not only for the detection of the pharmaceutically important drug Pen G, but also represents a general methodology for the detection of enzymes, eg PGA.

Conflicts of Interest

The authors declare no conflicts of interest.

Cite this paper

Tiwari, N. , Rathore, A. , Prabhune, A. and Kulkarni, S. (2010) Gold Nanoparticles for Colorimetric detection of hydrolysis of antibiotics by penicillin G acylase. Advances in Bioscience and Biotechnology, 1, 322-329. doi: 10.4236/abb.2010.14042.

References

[1] Laromaine A., Koh L., Murugesan M., Ulijn R.V. and Stevens, M.M. (2007), Protease triggered dispersion of nanoparticle assemblies. Journal of the Amemirican Chemical Society, 129(14), 4156-4157.
[2] Guarise, C., Pasquato, L., De Filippis, V. and Scrimin, P. (2006) Gold nanoparticles based protease assay. Proceedings of the National Academy of Sciences, 14, 3978- 3982.
[3] Zhang, X., Guo, Q. and Cui, D. (2009) Recent advances in Nanotechnology applied to biosensors. Sensors, 9(2), 1033-1053
[4] Li, Y., Schluesener, H.J. and Xu, S. (2010) Gold nanoparticles based biosensors. Gold Bulletin, 43 (1), 29.
[5] Liu, R., Teo, W., Tan, S., Feng, H., Padmanabhan, P. and Xing, B., (2010) Metallic nanoparticles bioassay for Enterobacter cloacae P99 β-lactamase activity and inhibitor screening. Analyst, 135(5), 1031.
[6] Batchelor, F.R., Chain, E.B., Hardy, T.L., Mansford, K.R.L. and Rolinson, G.N. (1961) 6-Aminopenicillanic acid. III. Isolation and purification. Proceedings of Royal SocietyB: Biological science, 154, 498-508.
[7] Bomstein, J. and Evans, W.G. (1965). Automated colorimetric determination of 6-Aminopenicillanic Acid in fermentation media. Analytical Chemistry, 37, 576-578.
[8] Sjoberg, G., Nathorst-Westfelt, L. and Ortengreen, B. (1967). Enzymatic hydrolysis of some penicillins and cephalosporins by Escherichia coli acylase. Acta Chemica Scandinavica, 21(2), 547-551.
[9] Baker, W.L. (1980). A note on the detection of penicillin acylase activity in Escherichia coli by the reaction of ampicillin with Buiret Reagent. The Journal of Applied Bacteriology, 49(2), 225-229.
[10] Mulvaney, P. (1996). Surface plasmon spectroscopy of nanosized metal particles. Langmuir, 12, 788-800.
[11] Riboh, J.C., Haes, A.J., McFarland, A.D., Yonzon, C.R. and Van Duyne, R.P. (2003). A nanoscale optical biosensor: real-time immunoassay in physiological buffer enabled by improved nanoparticle adhesion. The Journal of Physical Chemistry B., 107, 1772-1780.
[12] Nusz, G., Marinakos, S., Curry, A., Dahlin, A., Hook, F., Wax, A. and Chilkoti, A. (2008) Label-Free plasmonic detection of biomolecular binding by a single gold nanorod. Analytical Chemistry, 80(4), 984-989.
[13] Storhoff, J.J., Mucic, R.C., Letsinger, R.L., Mirkin, C.A.. (1997). Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles. Science, 277(5329), 1078-1080.
[14] Kalele, S.A., Ashtaputre, S.S., Hebalkar, N.Y., Gosavi, S.W., Deobagkar, D.N., Deobagkar, D.D. and Kulkarni, S.K. (2005). Optical detection of antibody using silica-silver core-shell particles. Chemical Physics Letters, 404(1-3), 136-141.
[15] Kalele, S.A., Kundu, A.A., Gosavi, S.W., Deobagkar, D.N., Deobagkar, D.D. and Kulkarni, S.K. (2006). Rapid detection of escherichia coli by using antibody-conjugated silver nanoshells. Small, 2(3), 335-338.
[16] Jena, B. and Raj, C. (2008) Optical sensing of biomedically important polyionic drugs using nano-sized gold particles. Biosensors and Bioelectronics, 23(8), 1285- 1290.
[17] Hossain, M., Huang, G., Kaneko, T. and Ozaki, Y. (2009) Characteristics of surface-enhanced Raman scattering and surface-enhanced fluorescence using a singleand a double layer gold nanostructure. Physical Chemistry Chemical Physics, 11(34), 7484.
[18] Kneipp, K., Kneipp, H., Itzkan, I., Dasari, R.R. and Feld, M.S. (1999). Ultrasensitive chemical analysis by raman spectroscopy. Chemical Review, 99(10), 2957-2975.
[19] Aslan, K., Lakowicz, J.R. and Geddes, C.D. (2005). Metal-enhanced fluorescence using anisotropic silver nanostructures: Critical progress to date. Analytical and Bioanalytical Chemistry, 382(4), 926-933.
[20] Liu, J. and Lu, Y. (2004). Adenosine-dependent assembly of aptazyme-functionalized gold nanoparticles and its application as a colorimetric biosensor. Analytical Chemistry, 76(6), 1627-1632.
[21] Ulijn, R.V. (2006). Enzyme-responsive materials: a new class of smart biomaterials. Journal of Materials Chemistry, 16, 2217-2225.
[22] Fischer, N.O., McIntosh, C.M., Simard, J.M. and Rotello, V.M. (2002). Supramolecular chemistry and self-assembly special feature: inhibition of chymotrypsin through surface binding using nanoparticle-based receptors. The Proceedings of the National Academy of Sciences, 99(8), 5018-5023.
[23] Storhoff, J.J., Lazarides, A.A., Mucic, R.C., Mirkin, C.A., Letsinger, R.L. and Schatz, G.C. (2000). What controls the optical properties of DNA-linked gold nanoparticle assemblies? Journal of American Chemical Society, 122, 4640.
[24] Brannigan, J.A., Dodson, G., Duggleby, H.J., Moody, P.C., Smith, J.L., Tomchick, D.R. and Murzin, A.G. (1995). A. protein catalytic framework with an N-terminal nucleophile is capable of self-activation. Nature, 378(6555), 416-419.
[25] Duggleby, H.J., Tolley, S.P., Hill, C.P., Dodson, E.J., Dodson, G.G. and Moody, P.C.E. (1995). Penicillin acylase has a single aminoacid catalytic center. Nature, 373, 264-268.
[26] Arroyo, M., de la Mata, I., Acebal, C. and Pilar Castill?n, M. (2003). Biotechnological application of penicillin acylases: State-of-the-art. Applied Microbiology and Biotechnology, 60(5), 507-514.
[27] Phadtare, S., Parekh, P., Gole, A., Patil, M., Pundle, A., Prabhune, A. and Sastry, M. (2002). Penicillin G acylase-fatty lipid biocomposite films show excellent catalytic activity and long term stability/reusability. Biotechnology Progress, 18(3), 483-488.
[28] Fadnavis, N.W., Sharfuddin, M. and Vadivel, S.K. (1999). Resolution of racemic 2-amino-1-butanol with immobilised penicillin G acylase. Tetrahedron Asymmetry, 10(23), 4495-4500.
[29] Van Langen, L.M., Oosthoek, N.P., Guranda, D.T., van Rantwijk, F., Svedas, V.K. and Sheldon, R.A. (2000). Penicillin acylase-catalyzed resolution of amines in aqueous organic solvents. Tetrahedron Asymmetry, 11(22), 4593.
[30] Shaikh, K., Talati, P.G. and Gang, D.M. (1973). Spectrophotometric method for the estimation of 6-aminopenicillanic acid. Antimicrobial Agents and Chemotherapy, 3, 194.
[31] Daumy, G.O., McColl, A.S. and Apostolakos, D. (1982). Repression of penicillin G acylase of Proteus rettgeri by tricarboxylic acid cycle intermediates. The Journal of Bacteriology, 152(1), 104.
[32] Tewari, Y.B. and Goldeberg, R.N. (1988). Thermodynamics of the conversion of penicillin G to phenylacetic acid and 6-aminopenicillanic acid. Biophysical Chemistry, 29(3), 245-252.
[33] Szewczuk, A., Siewinski, M. and Slowinska, R. (1980). Colorimetric assay of penicillin amidase activity using phenylacetyl-aminobenzoic acid as substrate. Analtical Biochemistry, 103(1), 166-169.
[34] Chiang, D. and Bennet, R.E. (1967). Purification and properties of penicillin amidase from bacillus megaterium. The Journal of Bacteriology, 93, 302-308.
[35] Kutzbach, C. and Rauenbusch, E. (1974). Preparation and general properties of crystalline penicillin acylase from Escherichia coli ATCC 11105. Hoppe-Seyler's Zeitschrift für physiologische Chemie, 355, 45-53.
[36] Balasingham, K., Warburton, D., Dunnill, P. and Lilly, M.D. (1972). The isolation and kinetics of penicillin amidase from Escherichia coli. Biochimica et Biophysica Acta, 276(1), 250-256.
[37] Shewale, G.J., Kumar, K.K. and Ambekar, G.R. (1987). Evaluation of determination of 6-aminopenicillanic acid by p-dimethyl aminobenzaldedyde. Biotechnological Techniques, 1, 69-72.
[38] Nikoobakht, B. and El-Sayed, M.A. (2007). Evidence for bilayer assembly of cationic surfactants on the surface of gold nanorods. Langmuir, 17, 6368-6374.

  
comments powered by Disqus

Copyright © 2018 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.