title: Interaction of small molecules with fungal laccase: A Surface Plasmon Resonance based study.
abstract: Laccases have a great potential for use in industrial and biotechnological applications. It has affinity towards phenolics and finds major applications in the field of bioremediation. Here, Surface Plasmon Resonance (SPR) as a biosensor with immobilized laccase on chip surface has been studied. Laccase was immobilized by thiol coupling method and compounds containing increasing number of hydroxyl groups were analyzed for their binding affinity at various concentrations in millimolar range. The small molecules like phloroglucinol (1.532×10(-8) M), crocin (3.204×10(-3) M), ascorbic acid (8.331×10(-8) M), kojic acid (6.411×10(-7) M) and saffron (3.466×10(-7) M) were studied and respective KD values are obtained. The results were also confirmed by inhibition assay and IC50 values were calculated. All these molecules showed different affinity towards laccase in terms of KD values. This method may be useful for preliminary screening and characterization of small molecules as laccase substrates, inhibitors or modulators of activity. This method will be useful for rapid screening of phenolics in waste water because of high sensitivity.
authors.affiliation: Department of Biotechnology, Shivaji University, Kolhapur, India.
authors.forename: Swati V
authors.initials: SV
authors.lastname: Surwase
authors.affiliation: Department of Biotechnology, Shivaji University, Kolhapur, India.
authors.forename: Sushama A
authors.initials: SA
authors.lastname: Patil
authors.affiliation: GE Healthcare Life Sciences, John F Welch Technology Centre, EPIP, Phase 2, Whitefield Road, Bangalore 560048, India.
authors.forename: Sistla
authors.initials: S
authors.lastname: Srinivas
authors.affiliation: Department of Biotechnology, Shivaji University, Kolhapur, India; Department of Biochemistry, Shivaji University, Kolhapur 416004, India. Electronic address: jpjbiochem@gmail.com.
authors.forename: Jyoti P
authors.initials: JP
authors.lastname: Jadhav
mesh_terms: D001205:Ascorbic Acid / Q000378:metabolism; D001673:Biodegradation, Environmental; D015374:Biosensing Techniques; D002338:Carotenoids / Q000378:metabolism; D003199:Computer Systems; D027622:Crocus / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D042845:Laccase / Q000378:metabolism*; D010696:Phloroglucinol / Q000378:metabolism; D011485:Protein Binding; D011753:Pyrones / Q000378:metabolism; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance; D014249:Trichophyton / Q000201:enzymology*
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D011753:Pyrones; D002338:Carotenoids; C011890:kojic acid; C029036:crocin; D010696:Phloroglucinol; D042845:Laccase; D001205:Ascorbic Acid
keywords: Affinity; IC(50); Surface Plasmon Resonance; T. versicolor laccase