Gallic acid interference on polyphenolic amperometric biosensing using Trametes versicolor laccase |
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| Institution: | 1. Division of Physical Science, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand;2. Center of Excellence for Innovation in Chemistry, Division of Physical Science, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand;3. Department of Chemistry and Center of Excellence in Materials Science and Technology, Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand;4. Environmental Science Research Center (ESRC), Faculty of Science, Chiang Mai University, Chiang Mai 50200, Thailand;5. Department of Biomedical Sciences and Biomedical Engineering, Faculty of Medicine, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand;1. School of Civil Engineering, Shijiazhuang Tiedao University, Shijiazhuang 050043, China;2. Key Laboratory of Roads and Railway Engineering Safety Control of Ministry of Education, Shijiazhuang Tiedao University, Shijiazhuang 050043, China;3. Graduate School, Shijiazhuang Tiedao University, Shijiazhuang 050043, China;4. School of Civil Engineering, Beijing Jiaotong University, Beijing 100044, China |
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| Abstract: | The present work reports the gallic acid (GA) interference on polyphenolic amperometric biosensing using Trametes versicolor laccase (TvLac). GA′ inhibitory effect on TvLac activity was investigated on the oxidation of caffeic acid (CA) by free TvLac and its immobilised form on modified polyethersulfone membrane (PES/TvLac), using spectrophotometric and amperometric biosensor detection methods. The results have indicated that GA presents inhibitory behaviour on TvLac activity in a concentration-dependent manner. The GA concentration leading to 50% activity lost, IC50, was determined to be 19.15 ± 0.11 μM and 5.11 ± 0.19 μM for free and immobilised enzyme, respectively. The results have also shown that GA exhibited a competitive and a mixed inhibition types on the TvLac activity for spectrophotometric and amperometric biosensor methods, respectively. Further GA′ and CA′ cyclic voltammetry studies have demonstrated that GA′ oxidation products interfered with CA′ redox reaction products. In fact, a decrease of the reduction current was observed at cyclic voltammograms of CA, when mixed with GA. Therefore, the GA′ interference on polyphenolic amperometric biosensing is the result of the combination of two factors: on one hand, we have the inhibitory enzymatic effect, and on the other, the reaction of GA′ oxidation products with the o-quinones obtained by the enzymatic oxidation of CA. Both gave rise to the amperometric signal decreasing effect. |
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