Biochemical Characterization and Potential for Textile Dye Degradation of Blue Laccase from Aspergillus ochraceus NCIM-1146

Amar A. Telke,Avinash A. Kadam,Sujit S. Jagtap,Jyoti P. Jadhav,Sanjay P. Govindwar 한국생물공학회 2010 Biotechnology and Bioprocess Engineering Vol.15 No.4

In our study, we produced intracellular blue laccase by growing the filamentous fungus Aspergillus ochraceus NCIM-1146 in potato dextrose broth. The enzyme was then purified 22-fold to a specific activity of 4.81 U/mg using anion-exchange and size exclusion chromatography. The molecular weight of purified laccase was estimated as 68 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme showed maximum substrate specificity toward 2,2'-Azinobis, 3-ethylbenzothiazoline-6-sulfonic acid than any other substrate. The optimum pH and temperature for laccase activity were 4.0 and 60ºC, respectively. The purified enzyme was stable up to 50ºC, and high laccase activity was maintained at pH 5.0 ~ 7.0. Laccase activity was strongly inhibited by sodium azide, EDTA, dithiothreitol, and L-cysteine. Purified laccase decolorized various textile dyes within 4 h in the absence of redox mediators. HPLC and FTIR analysis confirmed degradation of methyl orange. The metabolite formed after decolorization of methyl orange was characterized as p-N,N'-dimethylamine phenyldiazine using GCMS.

<i>Asparagus densiflorus</i> in a vertical subsurface flow phytoreactor for treatment of real textile effluent: A lab to land approach for <i>in situ</i> soil remediation

Watharkar, Anuprita D.,Kadam, Suhas K.,Khandare, Rahul V.,Kolekar, Parag D.,Jeon, Byong-Hun,Jadhav, Jyoti P.,Govindwar, Sanjay P. Elsevier 2018 Ecotoxicology and environmental safety Vol.161 No.-

<P><B>Abstract</B></P> <P>This study explores the potential of <I>Asparagus densiflorus</I> to treat disperse Rubin GFL (RGFL) dye and a real textile effluent in constructed vertical subsurface flow (VSbF) phytoreactor; its field cultivation for soil remediation offers a real green and economic way of environmental management. <I>A. densiflorus</I> decolorized RGFL (40 gm L<SUP>−1</SUP>) up to 91% within 48 h. VSbF phytoreactor successfully reduced American dye manufacture institute (ADMI), BOD, COD, Total Dissolved Solids (TDS) and Total Suspended Solids (TSS) of real textile effluent by 65%, 61%, 66%, 48% and 66%, respectively within 6 d. Oxidoreductive enzymes such as laccase (138%), lignin peroxidase (129%), riboflavin reductase (111%) were significantly expressed during RGFL degradation in <I>A. densiflorus</I> roots, while effluent transformation caused noteworthy induction of enzymes like, tyrosinase (205%), laccase (178%), veratryl oxidase (52%). Based on enzyme activities, UV–vis spectroscopy, FTIR and GC-MS results; RGFL was proposed to be transformed to 4-amino-3- methylphenyl (hydroxy) oxoammonium and N, N-diethyl aniline. Anatomical study of the advanced root tissue of <I>A. densiflorus</I> exhibited the progressive dye accumulation and removal during phytoremediation. HepG2 cell line and phytotoxicity study demonstrated reduced toxicity of biotransformed RGFL and treated effluent by <I>A. densiflorus,</I> respectively. On field remediation study revealed a noteworthy removal (67%) from polluted soil within 30 d.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Asparagus densiflorus</I> showed potential to transform disperse dye Rubin GFL. </LI> <LI> Vertical subsurface flow phytoreactor efficiently decolorized real textile effluent. </LI> <LI> Toxicity study confirmed the reduced toxicity of biotransformed dye and effluent. </LI> <LI> <I>In situ</I> soil remediation studies revealed a noteworthy removal of soil ADMI. </LI> <LI> Lab to land transfer of phytoremediation technology was successfully achieved. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Efficient decolorization and detoxification of textile industry effluent by <i>Salvinia molesta</i> in lagoon treatment

Chandanshive, Vishal V.,Rane, Niraj R.,Gholave, Avinash R.,Patil, Swapnil M.,Jeon, Byong-Hun,Govindwar, Sanjay P. Elsevier 2016 Environmental research Vol.150 No.-

<P><B>Abstract</B></P> <P> <I>Salvinia molesta</I>, an aquatic fern was observed to have a potential of degrading azo dye Rubine GFL up to 97% at a concentration of 100mg/L within 72h using 60±2g of root biomass. Both root as well as stem tissues showed induction in activities of the enzymes such as lignin peroxidase, veratryl alcohol oxidase, laccase, tyrosinase, catalase, DCIP reductase and superoxide dismutase during decolorization of Rubine GFL. FTIR, GC-MS, HPLC and UV–visible spectrophotometric analysis confirmed phytotransformation of the model dye into smaller molecules. Analysis of metabolites revealed breakdown of an azo bond of Rubine GFL by the action of lignin peroxidase and laccase and formation of 2-methyl-4-nitroaniline and N-methylbenzene-1, 4-diamine. Anatomical tracing of dye in the stem of <I>S. molesta</I> confirmed the presence of dye in tissues and subsequent removal after 48h of treatment. The concentration of chlorophyll pigments like chlorophyll a, chlorophyll b and carotenoid was observed during the treatment. Toxicity analysis on seeds of <I>Triticum aestivum</I> and <I>Phaseolus mungo</I> revealed the decreased toxicity of dye metabolites. <I>In situ</I> treatment of a real textile effluent was further monitored in a constructed lagoon of the dimensions of 7m×5m×2m (total surface area 35m<SUP>2</SUP>) using <I>S. molesta</I> for 192h. This large scale treatment was found to significantly reduce the values of COD, BOD<SUB>5</SUB> and ADMI by 76%, 82% and 81% considering initial values 1185, 1440mg/L and 950 units, respectively.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Macrophyte <I>S. molesta</I> showed a potential for textile dyes and effluent treatment </LI> <LI> A possible dye degradation pathway of Rubine GFL by <I>S. molesta</I> is proposed </LI> <LI> <I>S. molesta</I> in constructed lagoon treated 52,500L of textile effluent </LI> <LI> Phytotoxicity assay revealed less toxic nature of by-products after treatment </LI> <LI> Anatomical study of stem revealed entry and removal of Rubine GFL </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

<i>In situ</i> phytoremediation of dyes from textile wastewater using garden ornamental plants, effect on soil quality and plant growth

Chandanshive, Vishal V.,Kadam, Suhas K.,Khandare, Rahul V.,Kurade, Mayur B.,Jeon, Byong-Hun,Jadhav, Jyoti P.,Govindwar, Sanjay P. Elsevier 2018 CHEMOSPHERE - Vol.210 No.-

<P><B>Abstract</B></P> <P> <I>In situ</I> phytoremediation of dyes from textile wastewater was carried out in a high rate transpiration system ridges (91.4 m × 1.0 m) cultivated independently with <I>Tagetes patula, Aster amellus, Portulaca grandiflora and Gaillardia grandiflora</I> which reduced American Dye Manufacturers Institute color value by 59, 50, 46 and 73%, respectively within 30 d compared to dye accumulated in unplanted ridges. Significant increase in microbial count and electric conductivity of soil was observed during phytoremediation. Reduction in the contents of macro (N, P, K and C), micro (B, Cu, Fe and Mn) elements and heavy metals (Cd, As, Pb and Cr) was observed in the soil from planted ridges due to phyto-treatment. Root tissues of these plants showed significant increase in the specific activities of oxido-reductive enzymes such as lignin peroxidase, laccase, veratryl alcohol oxidase, tyrosinase and azo reductase during decolorization of textile dyes from soil. Anatomical studies of plants roots revealed the occurrence of textile dyes in tissues and subsequent degradation. A minor decrease in plant growth was also observed. Overall surveillance suggests that the use of garden ornamental plants on the ridges of constructed wetland for the treatment of dyes from wastewater along with the consortia of soil microbial flora is a wise and aesthetically pleasant strategy.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Ornamental plants at wetland ridges accumulated and degraded dyes from soil. </LI> <LI> Synergism between plants and microbes was involved in effective dye removal. </LI> <LI> Study revealed the entry and degradation of textile dyes in roots. </LI> <LI> Phytoremediation did not cause any severe toxicity on studied plants. </LI> <LI> The proposed treatment method was found to be aesthetically pleasant. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Phytobeds with <i>Fimbristylis dichotoma</i> and <i>Ammannia baccifera</i> for treatment of real textile effluent: An <i>in situ</i> treatment, anatomical studies and toxicity evaluation

Kadam, Suhas K.,Chandanshive, Vishal V.,Rane, Niraj R.,Patil, Swapnil M.,Gholave, Avinash R.,Khandare, Rahul V.,Bhosale, Amrut R.,Jeon, Byong-Hun,Govindwar, Sanjay P. Elsevier 2018 Environmental research Vol.160 No.-

<P><B>Abstract</B></P> <P> <I>Fimbristylis dichotoma, Ammannia baccifera</I> and their co-plantation consortium FA independently degraded Methyl Orange, simulated dye mixture and real textile effluent. Wild plants of <I>F. dichotoma</I> and <I>A. baccifera</I> with equal biomass showed 91% and 89% decolorization of Methyl Orange within 60h at a concentration of 50ppm, while 95% dye removal was achieved by consortium FA within 48h. Floating phyto-beds with co-plantation (<I>F. dichotoma</I> and <I>A. baccifera</I>) for the treatment of real textile effluent in a constructed wetland was observed to be more efficient and achieved 79%, 72%, 77%, 66% and 56% reductions in ADMI color value, COD, BOD, TDS and TSS of textile effluent, respectively. HPTLC, GC-MS, FTIR, UV–vis spectroscopy and activated oxido-reductive enzyme activities confirmed the phytotrasformation of parent dye in to new metabolites. T-RFLP analysis of rhizospheric bacteria of <I>F. dichotoma</I>, <I>A. baccifera</I> and consortium FA revealed the presence of 88, 98 and 223 genera which could have been involved in dye removal. Toxicity evaluation of products formed after phytotransformation of Methyl Orange by consortium FA on bivalves <I>Lamellidens marginalis</I> revealed less damage of the gills architecture when analyzed histologically. Toxicity measurement by Random Amplification of Polymorphic DNA (RAPD) technique revealed bivalve DNA banding pattern in treated Methyl Orange sample suggesting less toxic nature of phytotransformed dye products.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>F. dichotoma</I> L. and <I>A. baccifera</I> L. decolorized Methyl Orange and real textile dye effluent. </LI> <LI> Co-plantation of <I>F. dichotoma</I> L. and <I>A. baccifera</I> L. gave more efficient dye removal. </LI> <LI> Possible degradation pathways of Methyl Orange by all three systems are proposed. </LI> <LI> Effluents were treated note-worthily in floating phyto-beds by plants. </LI> <LI> Toxicity study on bivalve revealed less toxic nature of dye products. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Biological Conversion of Amino Acids to Higher Alcohols

El-Dalatony, Marwa M.,Saha, Shouvik,Govindwar, Sanjay P.,Abou-Shanab, Reda A.I.,Jeon, Byong-Hun Elsevier 2019 Trends in biotechnology Vol.37 No.8

<P>‘Higher’ alcohols, which contain more than two carbons, have a higher boiling point, higher cetane number, and higher energy density than ethanol. Blends of biodiesel and higher alcohols can be used in internal combustion engines as next-generation biofuels without any modification and are minimally corrosive over extensive use. Producing higher alcohols from biomass involves fermenting and metabolizing amino acids. In this review, we describe the pathways and regulatory mechanisms involved in amino acid bioprocessing to produce higher alcohols and the effects of amino acid supplementation as a nitrogen source for higher alcohol production. We also discuss the most recent approaches to improve higher alcohol production via genetic engineering technologies for three microorganisms: <I>Saccharomyces cerevisiae</I>, <I>Clostridium</I> spp., and <I>Escherichia coli</I>.</P> <P><B>Highlights</B></P> <P>Proteins are polymers of various amino acids, connected via peptide bonds and classified as a major feedstock for bioenergy production. Higher alcohols are high-density alternative fuels that increase the longevity of transportation fuels.</P> <P>Proteins have a significant role in the fermentation process by providing amino acids for the growth of microorganisms, and enhancement of sugar permeability, in carbohydrate-rich sources.</P> <P>Due to the environmental and economic advantages of recombinant DNA technology, fermentation is the most used process for industrial-scale alcohol production. Applying this technology to higher alcohols can significantly improve industrialization for advanced fuel production.</P> <P>Extraction techniques are used to separate and mitigate the toxicity of alcohols produced in the fermentation broth to maintain the microbial cell viability for longer.</P>

Acetoclastic methanogenesis led by <i>Methanosarcina</i> in anaerobic co-digestion of fats, oil and grease for enhanced production of methane

Kurade, Mayur B.,Saha, Shouvik,Salama, El-Sayed,Patil, Swapnil M.,Govindwar, Sanjay P.,Jeon, Byong-Hun Elsevier Applied Science 2019 Bioresource Technology Vol. No.

<P><B>Abstract</B></P> <P>Fats, oil and grease (FOG) are energy-dense wastes that substantially increase biomethane recovery. Shifts in the microbial community during anaerobic co-digestion of FOG was assessed to understand relationships between substrate digestion and microbial adaptations. Excessive addition of FOG inhibited the methanogenic activity during initial phase; however, it enhanced the ultimate methane production by 217% compared to the control. The dominance of Proteobacteria was decreased with a simultaneous increase in Firmicutes, Bacteriodetes, Synergistetes and Euryarchaeota during the co-digestion. A significant increase in <I>Syntrophomonas</I> (0.18–11%), <I>Sporanaerobacter</I> (0.14–6%) and <I>Propionispira</I> (0.02–19%) was observed during co-digestion, which substantiated their importance in acetogenesis. Among methanogenic Archaea, the dominance of <I>Methanosaeta</I> (94%) at the beginning of co-digestion was gradually replaced by <I>Methanosarcina</I> (0.52–95%)<I>.</I> The absence/relatively low abundance of syntrophic acetate oxidizers and hydrogenotrophic methanogens, and dominance of acetoclastic methanogens suggested that methane generation during co-digestion of FOG was predominantly conducted through acetoclastic pathway led by <I>Methanosarcina</I>.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The addition of fats, oil and grease enhanced ultimate methane production by 217%. </LI> <LI> Firmicutes, Bacteriodetes, Synergistetes and Euryarchaeota were greatly increased. </LI> <LI> Dominance of <I>Methanosaeta</I> was replaced by <I>Methanosarcina</I> at the end of digestion. </LI> <LI> Methane was predominantly generated through acetoclastic pathway by <I>Methanosarcina</I>. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Whole conversion of microalgal biomass into biofuels through successive high-throughput fermentation

El-Dalatony, Marwa M.,Salama, El-Sayed,Kurade, Mayur B.,Kim, Kyoung-Yeol,Govindwar, Sanjay P.,Kim, Jung Rae,Kwon, Eilhann E.,Min, Booki,Jang, Min,Oh, Sang-Eun,Chang, Soon Woong,Jeon, Byong-Hun Elsevier 2019 CHEMICAL ENGINEERING JOURNAL -LAUSANNE- Vol.360 No.-

<P><B>Abstract</B></P> <P>Microalgae represent a promising feedstock for biofuel production. However, the energy efficiency of microalgal pretreatment and conversion technologies needs to be improved to meet the economic viability. Herein, we introduce a novel integrated approach to achieve unprecedented energy conversion efficiency (46%) of microalgal biomass (<I>Chlamydomonas mexicana</I>). A successive high-throughput fermentation followed by transesterification were employed. This process provided a platform for maximum recovery of energy carriers from biomass utilization (89%). Serial fermentations were implemented for thorough utilization of the biomass constituents, starting with carbohydrate, followed by protein to derive ethanol (C2) and higher alcohols (C3–C5), respectively. Lipid was the dominant component after the previous fermentation, which was converted to biodiesel via transesterification process. Successive fermentations served as a bio-pretreatment to enhance the bioavailability of the leftover protein and lipid, which minimized the use of expensive and laborious methods for their extraction from the microalgal biomass. The proposed serial fermentation process would maximize the utilization of biomasses for biofuel production, with minimum leftover (11%).</P> <P><B>Highlights</B></P> <P> <UL> <LI> High throughput fermentations achieved 46% energy recovery from microalgae. </LI> <LI> Successive fermentations served as a biopretreatment to enhance the accessibility. </LI> <LI> 89% of biomass was converted into biofuels with less production of waste. </LI> <LI> Fermentation of the leftover protein produced 0.37 g-higher alcohols/g-amino acids. </LI> <LI> Transesterification of the remaining lipids produced 0.5 g-biodiesel/g-fatty acids. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Microbial acclimatization to lipidic-waste facilitates the efficacy of acidogenic fermentation

Saha, Shouvik,Jeon, Byong-Hun,Kurade, Mayur B.,Chatterjee, Pradip K.,Chang, Soon Woong,Markkandan, Kesavan,Salama, El-Sayed,Govindwar, Sanjay P.,Roh, Hyun-Seog Elsevier 2019 CHEMICAL ENGINEERING JOURNAL -LAUSANNE- Vol.358 No.-

<P><B>Abstract</B></P> <P>Lipidic-waste such as fat, oil, and grease (FOG) are promising substrates for achieving higher bioenergy yields. An inadequate presence of an effective microbiome in the anaerobic digesters is the bottleneck for the proper utilization of FOG. Gradual introduction of FOG (0.2%, 1.2%, and 2.4% as volatile solids) in acidogenic fermentation showed a significant improvement in hydrogen yield (72%), compared to the control, after 2.4% FOG loading. Volatile solid (VS) reduction reached up to 65% in high FOG reactors with complete removal of major unsaturated fatty acids. Removal of saturated fatty acids increased to 90%. Improvement in hydrogen productivity (46 mL d<SUP>−1</SUP>) occurred during step-wise loading of 2.4% FOG to the acclimatized microbiome. The metabolic shift toward carboxylic chain elongation produced C4 and C6 fatty acids at concentrations of 1.61 mM and 0.90 mM, respectively in the acidogenic reactors. High-throughput sequencing of 16S rRNA amplicons revealed that the acclimatization process enriched the phylum Firmicutes (90%), followed by Bacteroidetes (12%) and Cloacimonetes (11%). The abundance of these phyla and their respective genera confirmed their preeminent role in hydrolysis, hydrogenogenic acidogenesis, and carboxylic chain elongation to produce hydrogen and C4–C7 fatty acids. Thus, we suggest that the improvement of hydrogen production using a microbiome acclimatized to FOG, and simultaneous production of high value organics (C4–C7 fatty acids), could facilitate the greater efficacy of the acidogenic fermentation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Microbial acclimatization improved lipidic-waste utilization in acidogenic fermentation. </LI> <LI> Firmicutes, Bacteroidetes, and Cloacimonetes were abundant in the acclimatized microbiome. </LI> <LI> Hydrogen productivity enhanced to 46 mL d<SUP>−1</SUP> after acclimatization. </LI> <LI> Hydrogenogenic acidogenesis and carboxylic chain elongation produced C4–C7 fatty acids. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Co-planted floating phyto-bed along with microbial fuel cell for enhanced textile effluent treatment

Kadam, Suhas K.,Watharkar, Anuprita D.,Chandanshive, Vishal V.,Khandare, Rahul V.,Jeon, Byong-Hun,Jadhav, Jyoti P.,Govindwar, Sanjay P. Elsevier 2018 JOURNAL OF CLEANER PRODUCTION Vol.203 No.-

<P><B>Abstract</B></P> <P>A floating phytobed system based on plants of <I>Chrysopogon zizanioides</I> and <I>Typha angustifolia</I> (Consortium CT) was effective in the removal of Scarlet RR Dye (150 mg/L) and a textile effluent, with rates of 89 and 87%, respectively, within a 60-h period, which demonstrates a higher elimination rate than an individual plantation. In addition, the treatment of textile effluents with the floating phytobed linked to microbial fuel cells was enhanced in terms of color reduction, chemical oxygen demand, biological oxygen demand, total dissolved solids and total suspended solids up to 82, 75, 75, 67 and 70%, respectively. Moreover, it produced a power of 0.0769 W/m<SUP>2</SUP> at current density of 0.3846 A/m<SUP>2</SUP>. Terminal restriction length polymorphism community analysis documented 37 new genera which have a probable role in efficient treatment as well as power generation. Induction in the activities of oxidoreductase, high performance thin layer chromatography and gas chromatography-mass spectroscopy analyses of treated Scarlet RR dye confirmed the biotransformation. Toxicity evaluated on gill histology of <I>Lamellidens marginalis</I> and inter simple sequence repeat marker assessment confirmed the decreased toxicity of Scarlet RR after phyto-transformation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Chrysopogon zizanioides</I> and <I>Typha angustifolia</I> treated Scarlet RR and real effluent. </LI> <LI> Co-planted phyto-bed (FPb) gave efficient dye removal with energy generation. </LI> <LI> Removal of dyes improved note-worthily by FPb-Microbial Fuel Cells system. </LI> <LI> Degradation pathway of Scarlet RR by co-plantation system was proposed. </LI> <LI> Toxicity study on bivalve revealed less toxic nature of dye products. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>