Differential fate of metabolism of a disperse dye by microorganisms Galactomyces geotrichum and Brevibacillus laterosporus and their consortium GG-BL.

The present work aims to evaluate Brown 3 REL degrading potential of developed microbial consortium GG-BL using two microbial cultures, Galactomyces geotrichum MTCC 1360 (GG) and Brevibacillus laterosporus MTCC 2298 (BL). Microbial consortium GG-BL showed 100% decolorization of a dye Brown 3 REL, while individually G. geotrichum MTCC 1360 and B. laterosporus MTCC 2298 showed 26% and 86% decolorization under aerobic condition (shaking) respectively. Measurements of biochemical oxygen demand (BOD) (76%) and chemical oxygen demand (COD) (68%) were done after decolorization by consortium GG-BL. No induction in activities of oxidoreductive enzymes found in G. geotrichum while B. laterosporus showed induction of veratryl alcohol oxidase, Nicotineamide adenine dinucleotide-dichlorophenol indophenol (NADH-DCIP) reductase and riboflavin reductase indicating their role in dye metabolism. Consortium GG-BL showed induction in the activities of laccase, veratryl alcohol oxidase, tyrosinase, NADH-DCIP reductase and riboflavin reductase. Two different sets of induced enzymes from G. geotrichum and B. laterosporus work together in consortium GG-BL resulting in faster degradation of dye. The degradation of Brown 3 REL was analyzed using high performance thin layer chromatography (HPTLC), high performance liquid chromatography (HPLC), Fourier transform infrared spectroscopy (FT-IR) and gas chromatography mass spectroscopy (GC-MS). Phytotoxicity study revealed that metabolites formed after degradation was significantly less toxic in nature.

Phytodegradation of the triphenylmethane dye Malachite Green mediated by cell suspension cultures of Blumea malcolmii Hook.

Cell cultures of Blumea malcolmii Hook., developed in the laboratory, rapidly decolorized textile industry effluent along with a variety of dyes with diverse structural properties. Most rapid decolorization was observed in case of Malachite Green (93.41% decolorization within 24 h). The cells were capable of tolerating and degrading high concentrations of the dye, thus making them remarkable systems for phytoremediation studies. The enzymatic analysis during decolorization of Malachite Green showed the induction of enzymes such as laccase, veratryl alcohol oxidase and DCIP reductase indicating the involvement of these enzymes in the degradation of the dye. The cell cultures also mediated the remediation of textile industry effluent by bringing about a decrease in the BOD, COD and ADMI values of the effluent within 48 h. Phytotransformation was confirmed with the help of HPLC and the probable fate of metabolism of the dye was predicted with the help of GCMS analysis.

Biochemical characterization of laccase from hairy root culture of Brassica juncea L. and role of redox mediators to enhance its potential for the decolorization of textile dyes.

In vitro transgenic hairy root cultures provide a rapid system for physiological, biochemical studies and screening of plants for their phytoremediation potential. The hairy root cultures of Brassica juncea L. showed 92% decolorization of Methyl orange within 4 days. Out of the different redox mediators that were used to achieve enhanced decolorization, 2, 2'-Azinobis, 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) was found to be the most efficient. Laccase activity of 4.5 U mg(-1) of protein was observed in hairy root cultures of Brassica juncea L., after the decolorization of Methyl orange. Intracellular laccase produced by B. juncea root cultures grown in MS basal medium was purified up to 2.0 fold with 6.62 U mg(-1) specific activity using anion-exchange chromatography. Molecular weight of the purified laccase was estimated to be 148 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified enzyme efficiently oxidized ABTS which was also required for oxidation of the other tested substrates. The pH and temperature optimum for laccase activity were 4.0 and 40°C, respectively. The purified enzyme was stable up to 50°C and was stable in the pH range of 4.0-6.0. Laccase activity was strongly inhibited by sodium azide, EDTA, dithiothreitol and L: -cysteine. The purified enzyme decolorized various textile dyes in the presence of ABTS as an efficient redox mediator. These findings contribute to a better understanding of the enzymatic process involved in phytoremediation of textile dyes by using hairy roots.

A study on significant microbial interaction leading to decolorization and degradation of textile dye Rubine 3GP.

The present study evaluates an obligatory interaction between the yeast Saccharomyces cerevisiae NCIM 3312 and the bacterium Pseudomonas sp. strain BCH3 for the biodegradation of the dye Rubin 3GP (R3GP). No significant degradation of R3GP was observed either by Saccharomyces cerevisiae NCIM 3312 or by Pseudomonas sp. strain BCH3, when both the cultures were tested individually under their respective optimum medium conditions. However, when both of them were allowed to intermingle with each other, R3GP was found to be degraded within 72 h, with a steady increase in ß -1,3-glucanase, chitinase and protease activity in the culture supernatant; indicating the possible role of Pseudomonas sp. strain BCH3 in cell wall lysis of S. cerevisiae NCIM 3312. The present study elucidates a rare microbial interaction where the bacterium Pseudomonas sp. strain BCH3 utilizes lysed yeast cells as the sole source of nutrients for its own growth and subsequently performs decolorization and degradation of R3GP. Enzymatic status showed involvement of various oxidoreductive enzymes like lignin peroxidase, laccase, DCIP reductase and azo reductase, indicating their role in decolorization and degradation of R3GP. Degradation was confirmed using HPLC, FTIR analysis and the biochemical pathway of degradation was elucidated by using GC-MS analysis.

Studies on phytoremediation potentiality of Typhonium flagelliforme for the degradation of Brilliant Blue R.

In vitro culture plants of Typhonium flagelliforme were found to decolorize a variety of dyes, including Malachite Green, Red HE 8B, Methyl Orange, Reactive Red 2, Direct Red 5B (DR5B), Red HE 7B, Golden Yellow HER, Patent Blue, and Brilliant Blue R (BBR), to varying extents within 4 days. The enzymatic analysis of plant roots of aseptically raised plantlets performed before and after degradation of the dye BBR by these plantlets showed a significant induction in the activities of peroxidase, laccase, tyrosinase, and 2,6-dichlorophenol-indophenol reductase, which indicated the involvement of these enzymes in the metabolism of the dye. Comparative study of the enzyme status of the plants Typhonium flagelliforme and Blumea malcolmii during the degradation of DR5B and BBR showed marked variations in the enzyme profile with respect to the use of different sources of the enzyme. Phytoremediation of BBR using Typhonium flagelliforme was confirmed with high performance liquid chromatography and Fourier transform infrared spectroscopy analysis performed before and after the degradation of the dye. One of the products of the metabolism of the dye was identified as 4-(4-ethylimino-cyclohexa-2,5-dienylidinemethyl)-phenylamine with the aid of gas chromatography-mass spectroscopy (GC-MS) analysis. Significant decrease in the American Dye Manufacturer's Institute, biological oxygen demand, and chemical oxygen demand values of synthetic mixture of textile dyes and industrial effluent confirmed the decolorization and detoxification. Phytotoxicity studies also revealed the nontoxic nature of the metabolites of BBR.

Production of polyhydroxyhexadecanoic acid by using waste biomass of Sphingobacterium sp. ATM generated after degradation of textile dye Direct Red 5B.

The degradation of textile effluent using microorganisms has been studied extensively, but disposal of generated biomass after dye degradation is a serious problem. The isolated Sphingobacterium sp. ATM was found to decolorize dye Direct Red 5B (DR5B) and simultaneously it produced polyhydroxyhexadecanoic acid (PHD). The organism decolorized DR5B at 500mgl(-1) concentration within 24h of dye addition and gave optimum production of PHD. The medium contains carbon source as a molasses which was found to be more significant within all carbon sources used. The Nuclear Magnetic Resonance spectroscopy (NMR), Fourier Transform Infrared spectroscopy (FTIR) and Gas Chromatography-Mass Spectroscopy (GC-MS) characterization of polyhydroxyalkanoates obtained revealed the compound as a polyhydroxyhexadecanoic acid. The activity of PHA synthase was found more at 24h after dye addition. The enzymes responsible for dye degradation include veratrol oxidase, laccase, DCIP (2,6-dichlorophenol-indophenol) reductase, riboflavin reductase and azo reductase was found to be induced during decolorization process. The FTIR analysis of samples before and after decolorization of dye confirmed the biotransformation of DR5B. The GC-MS analysis of product obtained led to the identification of two metabolites after biotransformation of dye as p-amino benzenesulfonic acid and naphthalene-1-ol.

Biotechnological strategies for phytoremediation of the sulfonated azo dye Direct Red 5B using Blumea malcolmii Hook.

Tissue cultured shrub plants of Blumea malcolmii were found to decolorize Malachite green, Red HE8B, Methyl orange, Reactive Red 2 and Direct Red 5B at 20 mg L(-1) concentration to varying extent within three days. A significant induction in the activities of lignin peroxidase, tyrosinase, DCIP (2,6-dichlorophenol-indophenol) reductase, azoreductase and riboflavin reductase in the roots was observed during the decolorization of Direct Red 5B, which indicated their crucial role in the metabolism of the dye. HPLC (High Performance Liquid Chromatography) and FTIR (Fourier Transform Infrared Spectroscopy) analysis of the samples before and after decolorization of the dye confirmed the phytotransformation of Direct Red 5B. The GC-MS (Gas Chromatography Mass Spectroscopy) analysis of the products led us to the identification of three metabolites formed after phytotransformation of the dye as 4-(4-amino-phenylazo)-benzene sulfonic acid, 3-amino-7-carboxyamino-4-hydroxy-naphthalene-2-sulfonic acid and 7-carboxyamino-naphthalene-2-sulfonic acid.