Complete decolorization of the anthraquinone dye Reactive blue 5 by the concerted action of two peroxidases from Thanatephorus cucumeris Dec 1

Abstract

It is useful to identify and examine organisms that may prove useful for the treatment of dye-contaminated wastewater. Here, we report the purification and characterization of a new versatile peroxidase (VP) from the decolorizing microbe, Thanatephorus cucumeris Dec 1 (TcVP1). The purified TcVP1 after Mono P column chromatography showed a single band at 43 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino acid sequencing revealed that the N terminus of TcVP1 had the highest homology to Trametes versicolor MPG1, lignin peroxidase G (LiPG) IV, Bjerkandera adusta manganese peroxidase 1 (MnP1), and Bjerkandera sp. RBP (12 out of 14 amino acid residues, 86% identity). Mn(2+) oxidizing assay revealed that TcVP1 acted like a classical MnP at pH approximately 5, while dye-decolorizing and oxidation assays of aromatic compounds revealed that the enzyme acted like a LiP at pH approximately 3. TcVP1 showed particularly high decolorizing activity toward azo dyes. Furthermore, coapplication of TcVP1 and the dye-decolorizing peroxidase (DyP) from T. cucumeris Dec 1 was able to completely decolorize a representative anthraquinone dye, Reactive blue 5, in vitro. This decolorization proceeded sequentially; DyP decolorized Reactive blue 5 to light red-brown compounds, and then TcVP1 decolorized these colored intermediates to colorless. Following extended reactions, the absorbance corresponding to the conjugated double bond from phenyl (250-300 nm) decreased, indicating that aromatic rings were also degraded. These findings provide important new insights into microbial decolorizing mechanisms and may facilitate the future development of treatment strategies for dye wastewater.

Bacterial diversity and composition in major fresh produce growing soils affected by physiochemical properties and geographic locations

“…During decolorization of Orange T4LL, a significant induction in the activities of lignin peroxidase, tyrosinase and reductases (NADH-DCIP, azo and riboflavin) was observed (Dawkar et al, 2010). Dye degrading peroxidase degrades typical peroxidase substrates, but also degrades hydroxyl free anthraquinone, which is not transformed by other peroxidases (Marchis et al, 2011;Sugano et al, 2006). Likewise, a combination of lignin peroxidases and veratyl alcohol also enhances decolorization of azo and anthraquinone dyes (Joshi et al, 2010).…”

“…Redox mediators play an important role in oxidative degradation by Streptomyces laccases, primarily by facilitating the movement of electrons in the system (Gonzalez et al, 2009). Dye-degrading peroxidases are reported to degrade hydroxyl-free anthraquinone dyes (Marchis et al, 2011;Sugano et al, 2006). A combination of lignin peroxidases and veratyl alcohol was found to enhance the decolorization of azo and anthraquinone dyes (Joshi et al, 2010).…”

DyP, a Unique Dye-decolorizing Peroxidase, Represents a Novel Heme Peroxidase Family

“…DyP has several characteristics that distinguish it from all other peroxidases, including a particularly wide substrate specificity, a lack of homology to most other peroxidases, and the ability to function well under much lower pH conditions (3-3.2) compared with the other plant peroxidases (13,15). In terms of substrate specificity, DyP degrades the typical peroxidase substrates, but also degrades hydroxyl-free anthraquinone, which is not a substrate of other peroxidases (7,13,15). This is very important characteristic.…”

“…Using this strategy, yeast cytochrome c peroxidase (2) and chloroplast ascorbate peroxidase (3) were classified as class I peroxidases on account of their prokaryotic source. Representatives of class II include lignin peroxidase (LiP) 2 (4), manganese peroxidase (MnP) (5), and versatile peroxidase (6,7), whereas class III contains horseradish peroxidase (HRP) (8) and barley grain peroxidase (BGP) (9). This classification has been widely applied to most known peroxidases, with the exception of chloroperoxidase (CPO) isolated from the fungus, Caldariomyces fumago, which lacks primary structural homology with other peroxidases (10,11).…”

Wood production, wood technology, and biotechnological impacts

“…DyP family peroxidase bind heme but lack the heme-binding region conserved among the plant peroxidase superfamily including the fungal class II family , Faraco et al 2007). At least the T. cucumeris enzyme has also dye-decoulorising activities (Sugano et al 2006).…”