Laccase mediated diclofenac transformation and cytotoxicity assessment on mouse fibroblast 3T3-L1 preadipocytes

Abstract

Diclofenac is recently considered as one of the most devastating environmental pollutants, because of its biomagnification in the food chain which leads to potential harmful effects on non-targeted organisms. This study describes the optimized laccase mediated diclofenac transformation using response surface methodology (RSM) and cytotoxicity testing on mouse fibroblast 3T3-L1 preadipocytes. Three factors (laccase, syringaldehyde, reaction time) were used to optimize the diclofenac transformation. The optimum level of laccase, syringaldehyde and reaction time was found to be 1.91 U mL À1 , 187 mg and 51 min for diclofenac transformation (20 mg L À1 ). The cytotoxicity assessment on mouse fibroblast 3T3-L1 preadipocytes showed that a maximum of 67.9% cell death occurred at 72 h treatment with diclofenac (200 mg mL À1 ), while the cells treated with laccase treated diclofenac (LTD) showed less toxicity on the cells. These findings can be addressed for the removal of diclofenac toxicity.

Efficiency of Carbohydrate Additives on the Stability of Horseradish Peroxidase (HRP): HRP‐Catalyzed Removal of Phenol and Malachite Green Decolorization from Wastewater

“…RSM using CCD is a useful tool to determine the optimum level of variables and their interaction . In the present study, a second order polynomial equation relates the optimization of concentrations of galactose and guar gum to achieve maximum stabilization of HRP at 60°C.…”

“…For this investigation, a 2 3 fractional factorial CCD for three independent variables consisting of eight factorial points, six axial points, and six replicates at the center points leading to a total number of 20 experiments was used in this study. The number of experimental runs was calculated from the following equation : where N is the number of experiments required, k the number of variables, and x 0 is the number of central points. The range of variables investigated is presented in Tab.…”

“…RSM is a collection of statistical techniques widely applicable in optimization of various biochemical processes and helps to define the relationships between the response and independent variables [13,[24][25][26][27]. Regression model equations and operating conditions from appropriate experimental data are used to optimize the selected response [13,[24][25][26][27]. There are many designs available for fitting a second-order model [25][26][27].…”

“…Regression model equations and operating conditions from appropriate experimental data are used to optimize the selected response [13,[24][25][26][27]. There are many designs available for fitting a second-order model [25][26][27]. The most popular one is CCD method.…”

Ultrasound assisted enzymatic degradation of diclofenac sodium: Optimization of process parameters and kinetics

“…The cytotoxicity study reported by Sathishkumar et al demonstrates that diclofenac induced cell death, whereas, laccase treated diclofenac (LTD) did not induce considerable cell death. The cytotoxicity assessment was done on mouse fibroblast 3T3-L1 preadipocytes for which directly laccase treated diclofenac (LTD) has been used [36].…”