ABSTRACT
The accumulation of xenobiotic compounds in different environments interrupts the natural ecosystem and induces high toxicity in non-target organisms. Diclofenac is one of the commonly used pharmaceutical drugs that persist in the environment due to its low natural degradation rate and high toxicity. Therefore, this study aimed to isolate potential diclofenac-degrading bacteria, detect the intermediate metabolites formed, and determine the enzyme involved in the degradation process. Four bacterial isolates were selected based on their ability to utilize a high concentration of diclofenac (40 mg/L) as the sole carbon source. The growth conditions for diclofenac degradation were optimized, and bacteria were identified as Pseudomonas aeruginosa (S1), Alcaligenes aquatilis (S2), Achromobacter spanius (S11), and Achromobacter piechaudii (S18). The highest percentage of degradation was recorded (97.79 ± 0.84) after six days of incubation for A. spanius S11, as analyzed by HPLC. To detect and identify biodegradation metabolites, the GC-MS technique was conducted for the most efficient bacterial strains. In all tested isolates, the initial hydroxylation of diclofenac was detected. The cleavage step of the NH bridge between the aromatic rings and the subsequent cleavage of the ring adjacent to or in between the two hydroxyl groups of polyhydroxylated derivatives might be a key step that enables the complete biodegradation of diclofenac by A. piechaudii S18, as well as P. aeruginosa S1. Additionally, the laccase, peroxidase, and dioxygenase enzyme activities of the two Achromobacter strains, as well as P. aeruginosa S1, were tested in the presence and absence of diclofenac. The obtained results from this work are expected to be a useful reference for the development of effective detoxification bioprocesses utilizing bacterial cells as biocatalysts. The complete removal of pharmaceuticals from polluted water will stimulate water reuse, meeting the growing worldwide demand for clean and safe freshwater.
ABSTRACT
In order to improve the economic feasibility and environmental sustainability of microalgal bioethanol production, a nontoxic, copious agricultural waste, sugarcane bagasse aqueous extract (SBAE) was used for cultivating Nannochloropsis oculata microalga (NNO-1 UTEX Culture LB 2164) as potential sources of substitutes for traditional nutrition to reduce the costs in cultivation through acid digestion and enzymatic treatment before being fermented by Saccharomyces cerevisiae (NRRLY-2034). The primary target of this research was to find out the ethanol from hydrolysate of the defatted biomass of N. oculata grown mixotrophically on SBAE and CO2 as carbon sources. For acid hydrolysis (AH), the highest carbohydrate yield 252.84 mg/g DW has been obtained with 5.0% (v/v) H2SO4 at 121 °C for 15 min for defatted biomass cultivated mixotrophically on sugarcane bagasse aqueous extract (SBAE) regarding 207.41 mg/g DW for defatted biomass cultivated autotrophically (control treatment). Whereas, the highest levels of reducing sugars has been obtained with 4.0% (v/v) H2SO4 157.47±1.60 mg/g DW for defatted biomass cultivated mixotrophically compared with 135.30 mg/g DW for the defatted control treatment. The combination of acid hydrolysis 2.0% (v/v) H2SO4 followed by enzymatic treatment (AEH) increased the carbohydrate yields to 268.53 mg/g DW for defatted biomass cultivated mixotrophically on SBAE regarding 177.73 mg/g DW for the defatted control treatment. However, the highest levels of reducing sugars have been obtained with 3.0% (v/v) H2SO4 followed by enzyme treatment that gave 232.39±1.77 for defatted biomass cultivated mixotrophically on SBAE and 150.75 mg/g DW for the defatted control treatment. The sugar composition of the polysaccharides showed that glucose was the principal polysaccharide sugar (60.7-62.49%) of N. oculata defatted biomass. Fermentation of the hydrolysates by Saccharomyces cerevisiae for the acid pretreated defatted biomass samples gave ethanol yield of 0.86 g/L (0.062 g/g sugar consumed) for control and 1.17 g/L (0.069 g/g sugar consumed) for SBAE mixotrophic. Whereas, the maximum ethanol yield of 6.17±0.47 g/L (0.26±0.11 g/g sugar consumed) has been obtained with samples from defatted biomass grown mixotrophically (SBAE mixotrophic) pretreated with acid coupled enzyme hydrolysis.
