Expression and Characterization of a Dye-Decolorizing Peroxidase from Pseudomonas Fluorescens Pf0-1

Abstract

The consumption of dyes is increasing worldwide in line with the increase of population and demand for clothes and other colored products. However, the efficiency of dyeing processes is still poor and results in large amounts of colored effluents. It is desired to develop a portfolio of enzymes which can be used for the treatment of colored wastewaters. Herein, we used genome sequence information to discover a dye-decolorizing peroxidase (DyP) from Pseudomonas fluorescens Pf-01. Two genes putatively encoding for DyPs were identified in the respective genome and cloned for expression in Escherichia coli, of which one (PfDyP B2) could be overexpressed as a soluble protein. PfDyP B2 shows some typical features known for DyPs which includes the ability to convert dyes at the expense of hydrogen peroxide. Interestingly, t-butyl hydroperoxide could be used as an alternative substrate to hydrogen peroxide. Immobilization of PfDyP B2 in calcium-alginate beads resulted in a significant increase in stability: PfDyP B2 retains 80% of its initial activity after 2 h incubation at 50 °C, while the soluble enzyme is inactivated within minutes. PfDyP B2 was also tested with aniline and ethyl diazoacetate as substrates. Based on GC-MS analyses, 30% conversion of the starting material was achieved after 65 h at 30 °C. Importantly, this is the first report of a DyP-catalyzed insertion of a carbene into an N-H bond.

Characterization of Two Hydrogen Peroxide Resistant Peroxidases from Rhodococcus opacus 1CP

“…It is also noteworthy to investigate if DypB does play a role on lignin degradation. Moreover, a study [69] showed that the dye-decolorizing peroxidase Pf DyP B2 from Pseudomonas fluorescences Pf0-1 can insert carbene into an N-H bond. Investigation of these non-natural reactions can be a nice addition to the enzymatic reactions that these dye-decolorizing peroxidases can do.…”

Cultivation of Mushrooms and Their Lignocellulolytic Enzyme Production Through the Utilization of Agro-Industrial Waste

“…Enzyme technology possesses great potential to reduce environmental pollution and offers potential benefits in the comprehensive utilization of lignocellulosic biomass. Lignocellulolytic enzymes have received attention because of their potential applications in various agro-industrial bioprocesses, such as the conversion of hemicellulosic biomass to fuels and chemical production, the clarification of juices, the green processing of certain foods and beverages, the enhancement of animal digestibility in feedstock, the delignification of paper and pulp, the improvement of fabric properties in the textile industry and waste utilization [277][278][279]. Cellulase is widely used in the textile and laundry detergent industries as it can play a part in the hydrolysis of cellulose and improve fabric properties for the textile industry and for cleaning textiles in the laundry detergent industry [154,280].…”

“…The food and beverage processing industries have used cellulase for the hydrolysis of cellulose during the drying of coffee beans and for the extraction of fruits and vegetables in juice production [281,282]. Cellulase, α-l-arabinofuranosidases and other glycosidases have also been used in brewery and wine production [213,277]. The enzymatic hydrolysis of grapes utilizes α-l-arabinofuranosidases and other glycosidases to enhance the flavor of wine by the release of free terpenols, an important aspect in the development of the aroma in wine.…”

“…Cellulases, xylanase and other hemicellulases are commonly used enzymes to assist in pulp bleaching for the reduction of environmental pollution loads [283]. Cellulases are used to improve the performance of dissolved pulp [277]. Additionally, α-l-arabinofuranosidases enhance the delignification of pulp in the bleaching process as it can cleave the arabionose side chain that inhibits the action of xylanase [213].…”

Efficient Degradation of 2-Mercaptobenzothiazole and Other Emerging Pollutants by Recombinant Bacterial Dye-Decolorizing Peroxidases

“…Bacterial DyPs are increasingly being discovered and are the most abundant members of the putative DyP family (PeroxiBase; accessed on 9 March 2021). The continuous advancement in recombinant DNA technology has improved the potential for the successful and cheap production of active recombinant DyPs in Escherichia coli systems [ 29 , 30 , 31 ]. It is interesting to note that, unlike their eukaryotic counterparts, such as SBP and HRP, which are very difficult to produce in active forms using bacterial expression systems, recombinant bacterial and fungal DyPs appear to be reasonably well expressed in E. coli .…”