Novel laccase from Xylaria polymorpha and its efficiency in the biotransformation of pharmaceuticals: Optimization of operational conditions, comparative effect of redox-mediators and toxicity studies.

The promising potentials of biocatalytic treatment processes in the removal of micropollutants whilst eliminating health and environmental hazards have attracted great attention in recent years. This current work investigated the biotransformation efficiency of a novel laccase from Xylaria polymorpha (XPL) in comparison with commercial laccases from Trametes versicolor (TVL) and Aspergillus sp. (ASL). XPL exhibited better oxidation performance (95.7%) on AMX than TVL (92.8%) and ASL (90.5%). Optimization of operational conditions revealed that AMX was best oxidized at pH 5, temperature (30 °C), and concentration (1.0 mg L-1). The investigation carried out to determine the effect of redox mediators revealed violuric acid (VLA) as the best redox mediator. The laccase stability experiments elucidated that the oxidation of AMX is time and mediator concentration dependent with ABTS exhibiting highest deactivation of XPL active sites. Two metabolic products; amoxicillin penilloic acid and 5-hydroxy-6-(4-hydroxyphenyl)- 3-(1,3-thiazolidin-2-yl)piperazin-2-one of AMX were obtained through Liquid Chromatography-Mass Spectrometry (LC-MS) analyses. The toxicity assessments carried out after oxidation of AMX by XPL showed 94% and 97% reduced toxicity on Artemia salina and Aliivibrio fischeri respectively. The study further underscored the efficiency of biocatalytic-mediator technology in the transformation of complex micropollutants into less toxic substances in an eco-friendly way.

Impact of redox-mediators in the degradation of olsalazine by marine-derived fungus, Aspergillus aculeatus strain bpo2: Response surface methodology, laccase stability and kinetics.

The indiscriminate disposal of olsalazine in the environment poses a threat to human health and natural ecosystems because of its cytotoxic and genotoxic nature. In the present study, degradation efficiency of olsalazine by the marine-derived fungus, Aspergillus aculeatus (MT492456) was investigated. Optimization of physicochemical parameters (pH. Temperature, Dry weight) and redox mediators {(2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), p-Coumaric acid and 1-hydroxybenzotriazole (HOBT)} was achieved with Response Surface Methodology (RSM)-Box-Behnken Design (BBD) resulting in 89.43% removal of olsalazine on 7th day. The second-order polynomial regression model was found to be statistically significant, adequate and fit with p < 0.0001, F value=41.87 and correlation coefficient (R2=0.9826). Biotransformation was enhanced in the redox mediator-laccase systems resulting in 99.5% degradation of olsalazine. The efficiency of ABTS in the removal of olsalazine was more pronounced than HOBT and p-Coumaric acid in the laccase-mediator system. This is attributed to the potent nature of the electron transfer mechanism deployed during oxidation of olsalazine. The pseudo-second-order kinetics revealed that the average half-life (t1/2) and removal rates (k1) increases with increasing concentrations of olsalazine. Michaelis-Menten kinetics affirmed the interaction between laccase and olsalazine under optimized conditions with maximum removal rate, Vmax=111.11 hr-1 and half-saturation constant, Km=1537 mg L-1. At the highest drug concentration (2 mM); 98%, 95% and 93% laccase was remarkably stabilized in the enzyme-drug degradation system by HOBT, ABTS and p-Coumaric acid respectively. This study further revealed that the deactivation of laccase by the redox mediators is adequately compensated with enhanced removal of olsalazine.

Biodegradation of fluorene by the newly isolated marine-derived fungus, Mucor irregularis strain bpo1 using response surface methodology.

Fluorene, a low molecular weight polycyclic aromatic hydrocarbon (PAH), is of immense environmental interest because of its carcinogenicity, teratogenicity, mutagenicity, toxicity and persistence to microbial degradation. Existentially, there is paucity of information on PAH degradation by fungi isolated from marine environment. Therefore, this study investigated fluorene degradation efficiency of marine derived filamentous fungus, Mucor irregularis strain bpo1 (GenBank Accession Number: MK373020). Response Surface Methodology (RSM) using Box-Behnken Design (BBD) was successfully deployed in the optimization of process parameters (pH-7, temperature-32.5 °C, substrate concentration-100 mg L-1 and dry weight-2 g) resulting in 81.50% fluorene degradation on 5th day. The design and regression model were found to be statistically significant, adequate and appropriate with p < 0.0001, F value= 202.39, and predicted coefficient of determination (R2 =0.9991). Optimization of the vital constituents of the mineral salt medium (MSM) used for the study using RSM-Central Composite Design (CCD) resulted in 79.80% fluorene degradation rate. Enhanced fluorene degradation efficiency (82.50%) was recorded when the optimized process variables were subjected to growth-linked validation experiments. The enzyme activities revealed 87%, 59% and 31% induction of laccase, manganese peroxidase and lignin peroxidase respectively. Four metabolites; 9H-fluoren-9-one, benzene-1,2-dicarboxylic acid, 2-hydroxybenzoic acid and phenol obtained after the experiment were characterized and confirmed with GC-MS analysis. The findings revealed the promising potentials of M. irregularis in PAH degradation and by extension green remediation technology.