Ultrarobust stable ABTS radical cation prepared using Spore@Cu-TMA biocomposites for antioxidant capacity assay

Herein, spore@Cu-trimesic acid (TMA) biocomposites were prepared by self-assembling Cu-based metal-organic framework on the surface of Bacillus velezensis spores. The laccase-like activity of spore@Cu-TMA biocomposites was enhanced by 14.9 times compared with that of pure spores due to the reaction of Cu²⁺ ions with laccase on the spore surface and the microporous structure of Cu-TMA shell promoting material transport and increasing substrate accessibility. Spore@Cu-TMA rapidly oxidized and transformed 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) into ABTS^●+ without using H₂O₂. Under optimum conditions, the ABTS^●+ could be stored for 21 days at 4 °C and 7 days at 37 °C without the addition of any stabilizers, allowing for the large-scale preparation and long-term storage of ABTS^●+. The ultrarobust stable ABTS^●+ obtained with the use of Cu-TMA could effectively reduce the "back reaction" by preventing the leaching of the metabolites released by the spores. On the basis of these findings, a rapid, low-cost, and eco-friendly colorimetric platform was successfully developed for the detection of antioxidant capacity. Determination of antioxidant capacity for several antioxidants such as caffeic acid, glutathione, and Trolox revealed their corresponding limits of detection at 4.83, 8.89, and 7.39 nM, respectively, with linear ranges of 0.01-130, 0.01-140, and 0.01-180 μM, respectively. This study provides a facile way to prepare ultrarobust stable ABTS^●+ and presents a potential application of spore@Cu-TMA biocomposites in food detection and bioanalysis.