Sayin, Serkan/0000-0003-0518-3208 WOS: 000405114700014 Aim: The development of calixarene based phenol biosensor. Methods: This study describes the application of a calixarene derivative, 5,17-diamino-25,27-bis(3-thiol-1- oxypropane)-26,28-dihydroxycalix[4] arene (HS-Calix-NH2) which has both amino and thiol functionalities, in the practical surface modifications for biomolecule binding. The structure of HS-Calix-NH2 allows easy interaction with Au surface and one-step biomolecule immobilization. Self-assembled monolayers (SAMs) of p-amino-functionalized mercaptoalkylcalixarene (HS-Calix-NH2) were formed onto the Au electrode. Then, Laccase (Lac) enzyme was immobilized onto the modified surface by crosslinking with glutaraldehyde (GA). Resulted electrode (HS-Calix-NH2/Lac) was used for the electrochemical analysis of phenolic compounds at -50 mV. Results: The linearity was observed in the range of 0.1-100 mu M and 1.0-100 mu M for catechol and phenol, respectively. The potential use of the biosensor was investigated for phenol analysis in artificial samples which simulate the industrial waste water, which is highly acidic and composed of concentrated salt, without needing any sample pre-treatment step. Conclusion: The prepared Lac biosensor has a potential for rapid, selective and easy detection of phenolic contaminations in samples. TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TUBITAK-2209]; European Union through the COST Action "Supramolecular photocatalytic water splitting (PERSPECT-H2O)" [CM1202]; Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [113T022]; Turkish Academy of Sciences-Outstanding Young Scientists Award Program (TUBA-GEBIP)Turkish Academy of Sciences Dr. D.O. Demirkol thanks to The Turkish Academy of Sciences-Outstanding Young Scientists Award Program (TUBA-GEBIP 2015). The authors also would like to thank TUBITAK for its Supporting Research Projects Program for the University Students at undergraduate level (Program Number TUBITAK-2209). This study was partially supported by the European Union through the COST Action CM1202 "Supramolecular photocatalytic water splitting (PERSPECT-H2O)" and the Scientific and Technological Research Council of Turkey (TUBITAK Grant Number 113T022).