, Volume 13, Issue 4, pp 1409–1415 | Cite as

Laccase Activity Assay Using Surface Plasmon Resonance Band of Gold Nanoparticles Formed by Dopamine

  • Kh. Pashangeh
  • M. R. Hormozi-Nezhad
  • M. Akhond
  • G. AbsalanEmail author


A simple, fast, and sensitive colorimetric technique for determination of laccase activity using dopamine (DA) induced growth of colloidal gold nanoparticles is proposed. It was found that the reduction of AuCl4 to colloidal gold nanoparticles (AuNPs) by dopamine (DA) in the presence of citrate ion as stabilizing agent produced a very intense surface plasmon resonance peak of AuNPs at 530 nm. As the activity of laccase (at fixed concentration of DA) increases, the oxidation of DA to dopamine-o-quinone (DOQ) is enhanced. The latter product could not act as the reducing agent for the reduction of AuCl4 to AuNPs. So, as the activity of laccase increases, the absorbance characteristic to the plasmon of the AuNPs at 530 nm is diminished. This reductive mechanism of the plasmon absorbance of the AuNPs allows the quantitative colorimetric assay for laccase activity. The linear range of the method is 0.1–10 U ml−1 laccase. The developed method has been applied to assay laccase activity in 12 samples per hour.

Graphical Abstract


Laccase Dopamine AuNPs formation Laccase activity Colorimetric assay 



The authors wish to acknowledge the support of this work by the Sharif University of Technology Research Council as well as the Shiraz University Research Council.


  1. 1.
    Singh G, Capalash N, Goel R, Sharma P (2007) A pH-stable laccase from alkali-tolerant γ-proteobacterium JB: purification, characterization and indigo carmine degradation. Enzym Microb Technol 41:794–799CrossRefGoogle Scholar
  2. 2.
    Yuan J, Guo W, Wang E (2008) Utilizing a CdTe quantum dots−enzyme hybrid system for the determination of both phenolic compounds and hydrogen peroxide. Anal Chem 80:1141–1145CrossRefPubMedGoogle Scholar
  3. 3.
    Gianfreda L, Iamarino G, Scelza R, Rao MA (2006) Oxidative catalysts for the transformation of phenolic pollutants: a brief review. Biocatal Biotransform 24:177–187CrossRefGoogle Scholar
  4. 4.
    Shamsipur M, Shanehasz M, Khajeh KH, Mollania N, Kazemi SH (2012) A novel quantum dot-laccase hybrid nanobiosensor for low level determination of dopamine. Analyst 137:5553–5559CrossRefPubMedGoogle Scholar
  5. 5.
    Thurston CF (1994) The structure and function of fungal laccases. Microbiology 140:19–26CrossRefGoogle Scholar
  6. 6.
    Bertrand T, Jolivalt C, Briozzo P, Caminade E, Joly N, Madzak C, Mougin C (2002) Crystal structure of a four-copper laccase complexed with an Arylamine: insights into substrate recognition and correlation with kinetics. Biochemistry 41:7325–7333CrossRefPubMedGoogle Scholar
  7. 7.
    Solomon EI, Sundaram UM, Machonkin TE (1996) Multicopper oxidases and oxygenases. Chem Rev 96:2563–2605CrossRefPubMedGoogle Scholar
  8. 8.
    Sharma P, Goel R, Capalash N (2007) Bacterial laccases. World J Microbiol Biotechnol 23:823–832CrossRefGoogle Scholar
  9. 9.
    Mogharabi M, Faramarzi MA (2014) Laccase and laccase-mediated systems in the synthesis of organic compounds. Adv Synth Catal 356:897–927CrossRefGoogle Scholar
  10. 10.
    Xu F (2005) Applications of oxidoreductases: recent progress. Ind Biotech 1:38–50CrossRefGoogle Scholar
  11. 11.
    Civjan N (2012) Chemical biology: approaches to drug discovery and development to targeting disease. John Wiley & SonsGoogle Scholar
  12. 12.
    Guo Z, Seol ML, Kim MS, Ahn JH, Choi YK, Liu JH, Huang XJ (2013) Sensitive and selective electrochemical detection of dopamine using an electrode modified with carboxylated carbonaceous spheres. Analyst 138:2683–2690CrossRefPubMedGoogle Scholar
  13. 13.
    Kim YR, Bong S, Kang YJ, Yang Y, Mahajan RK, Kim JS, Kim H (2010) Electrochemical detection of dopamine in the presence of ascorbic acid using graphene modified electrodes. Biosens Bioelectron 25:2366–2369CrossRefPubMedGoogle Scholar
  14. 14.
    Peik-See T, Pandikumar A, Nay-Ming H, Hong-Ngee L, Sulaiman Y (2014) Simultaneous electrochemical detection of dopamine and ascorbic acid using an iron oxide/reduced graphene oxide modified glassy carbon electrode. Sensors 14:15227–15243CrossRefPubMedGoogle Scholar
  15. 15.
    Baron R, Zayats M, Willner I (2005) Dopamine-, L-DOPA-, adrenaline-, and noradrenaline-induced growth of au nanoparticles: assays for the detection of neurotransmitters and of tyrosinase activity. Anal Chem 77:1566–1571CrossRefPubMedGoogle Scholar
  16. 16.
    Link S, El-Sayed MA (1999) Spectral properties and relaxation dynamics of surface plasmon electronic oscillations in gold and silver nanodots and nanorods. J Phys Chem B 103:8410–8426CrossRefGoogle Scholar
  17. 17.
    El-Sayed MA (2001) Some interesting properties of metals confined in time and nanometer space of different shapes. Acc Chem Res 34:257–264CrossRefPubMedGoogle Scholar
  18. 18.
    Templeton AC, Wuelfing WP, Murray RW (2000) Monolayer-protected cluster molecules. Acc Chem Res 33:27–36CrossRefPubMedGoogle Scholar
  19. 19.
    Liao S, Qiao Y, Han W, Xie Z, Wu ZH, Shen G, Yu R (2012) Acetylcholinesterase liquid crystal biosensor based on modulated growth of gold nanoparticles for amplified detection of acetylcholine and inhibitor. Anal Chem 84:45–49CrossRefPubMedGoogle Scholar
  20. 20.
    Zhang Y, Peng H, Huang W, Zhou Y, Yan D (2008) Facile preparation and characterization of highly antimicrobial colloid Ag or Au nanoparticles. Colloid Interface Sci 325:371–376CrossRefGoogle Scholar
  21. 21.
    Perez Y, Mann E, Herradon B (2011) Preparation and characterization of gold nanoparticles capped by peptide–biphenyl hybrids. Colloid Interface Sci 359:443–453CrossRefGoogle Scholar
  22. 22.
    El-Batal AI, ElKenawy NM, Yassin AS, Amin MA (2015) Laccase production by Pleurotus ostreatus and its application in synthesis of gold nanoparticles. Biotechnol Reports 5:31–39CrossRefGoogle Scholar
  23. 23.
    Faramarzi MA, Forootanfar H (2011) Biosynthesis and characterization of gold nanoparticles produced by laccase from Paraconiothyrium variabile. Colloid Surf 87:23–27CrossRefGoogle Scholar
  24. 24.
    XIE X, XU W, LIU X (2012) Improving colorimetric assays through protein enzyme-assisted gold nanoparticle amplification. Acc Chem Res 45:1511–1520CrossRefPubMedGoogle Scholar
  25. 25.
    Saha K, Agasti SS, Kim C, Li X, Rotello VM (2012) Gold nanoparticles in chemical and biological sensing. Chem Rev 112:2739–2779CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Hormozi-Nezhad MR, Tashkhourian J, Khodaveisi J (2010) Sensitive spectrophotometric detection of dopamine, levodopa and adrenaline using surface plasmon resonance band of silver nanoparticles. J Iran Chem Soc 7:83–91CrossRefGoogle Scholar
  27. 27.
    Sun Y, Xia Y (2003) Gold and silver nanoparticles: a class of chromophores with colors tunable in the range from 400 to 750 nm. Analyst 128:686–691CrossRefPubMedGoogle Scholar
  28. 28.
    McFarland AD, Van Duyne RP (2003) Single silver nanoparticles as real-time optical sensors with zeptomole sensitivity. Nano Lett 3:1057–1062CrossRefGoogle Scholar
  29. 29.
    Alberts JF, Gelderblom WCA, Botha A, Vanzyl WH (2009) Degradation of aflatoxin B1 by fungal laccase enzymes. Int J Food Microbiol 135:47–52CrossRefPubMedGoogle Scholar
  30. 30.
    Lu L, Zhao M, Zhang BB, Yu SY, Bian XJ, Wang W, Wang Y (2007) Purification and characterization of laccase from Pycnoporus sanguineus and decolorization of an anthraquinone dye by the enzyme. Appl Microbiol Biotechnol 74:1232–1239CrossRefPubMedGoogle Scholar
  31. 31.
    Simpson RJ (2003) Proteins and proteomics: a laboratory manual, first edn. CSHL Press, New York, pp 857–859Google Scholar
  32. 32.
    Shakibaie M, Forootanfar H, Mollazadeh-Moghaddam K, Bagherzadeh Z, Nafissi-Varcheh N, Shahverdi AR, Faramarzi MA (2010) Green synthesis of gold nanoparticles by the marine microalga Tetraselmis suecica. Biotechnol Appl Biochem 57:71–75CrossRefPubMedGoogle Scholar
  33. 33.
    Kalishwaralal K, Gopalram S, Vaidyanathan R, Deepak V, Pandian SRK, Gurunathan S (2010) Optimization of α-amylase production for the green synthesis of gold nanoparticles. Colloid Surf B 77:174–180CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • Kh. Pashangeh
    • 1
  • M. R. Hormozi-Nezhad
    • 2
    • 3
  • M. Akhond
    • 1
  • G. Absalan
    • 1
    Email author
  1. 1.Professor Massoumi Laboratory, Department of Chemistry, Faculty of SciencesShiraz UniversityShirazIran
  2. 2.Department of ChemistrySharif University of TechnologyTehranIran
  3. 3.Institute for Nanoscience and NanotechnologySharif University of TechnologyTehranIran

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