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        Toxicity of sulfamethazine and sulfamethoxazole and their removal by a green microalga, <i>Scenedesmus obliquus</i>

        Xiong, Jiu-Qiang,Govindwar, Sanjay,Kurade, Mayur B.,Paeng, Ki-Jung,Roh, Hyun-Seog,Khan, Moonis Ali,Jeon, Byong-Hun Pergamon Press 2019 Chemosphere Vol. No.

        <P><B>Abstract</B></P> <P>A comprehensive ecotoxicological evaluation of a sulfamethazine (SMZ) and sulfamethoxazole (SMX) mixture was conducted using an indicator microalga, <I>Scenedesmus obliquus</I>. The toxicological effects of this mixture were studied using microalgal growth patterns, biochemical characteristics (total chlorophyll, carotenoid, carbohydrate, fatty acid methyl ester), and elemental and Fourier-transform infrared spectroscopy analyses. The 96-h half maximal effective concentration (EC<SUB>50</SUB>) of the SMZ and SMX mixture was calculated to be 0.15 mg L<SUP>−1</SUP> according to the dose-response curves obtained. The chlorophyll content decreased with elevated SMZ and SMX concentrations, while the carotenoid content initially increased and then decreased as concentration raised. The unsaturated fatty acid methyl esters (FAMEs) content was enhanced with higher SMZ and SMX concentrations, while that of saturated FAMEs simultaneously decreased due to SMZ and SMX stress. Elemental analyses showed an improved percentage of nitrogen and sulfur in the microalgal biomass as SMZ and SMX concentrations increased. The microalga <I>S. obliquus</I> was shown to biodegrade the chemicals tested and removed 31.4–62.3% of the 0.025–0.25 mg SMZ L<SUP>−1</SUP> and 27.7–46.8% of the 0.025–0.25 mg SMX L<SUP>−1</SUP> in the mixture after 12 days of cultivation. The greater biodegradation observed at higher SMZ and SMX concentrations indicates that microalgal degradation of SMZ and SMX could act as an efficient adaptive mechanism to antibiotics.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>S. obliquus</I> can withstand high doses of SMZ and SMX. </LI> <LI> EC<SUB>50</SUB> of SMZ, SMX and their mixture for <I>S. obliquus</I> was 1.23, 0.12, and 0.15 mg L<SUP>−1</SUP>. </LI> <LI> <I>S. obliquus</I> removed 62.3 and 46.8% of SMZ and SMX, respectively. </LI> <LI> A greater biodegradation was observed in higher SMZ and SMX concentration. </LI> </UL> </P>

      • Insights into microalgae mediated biodegradation of diazinon by Chlorella vulgaris: Microalgal tolerance to xenobiotic pollutants and metabolism

        Kurade, M.B.,Kim, J.R.,Govindwar, S.P.,Jeon, B.H. Elsevier B.V 2016 Algal research Vol.20 No.-

        <P>Diazinon is one of the most widely used organophosphorus insecticides for agricultural activities, and it is highly toxic to mammals and other non-target organisms. The present study demonstrated the effective removal of diazinon from the aqueous phase by a freshwater, green microalga, Chlorella vulgaris. Among the four screened species (Scenedesmus obliquus, Chlamydomonas mexicana, Chlorella vulgaris and Chlamydomonas pitschmannii), C. vulgaris showed the highest removal capacity (94%) of diazinon at 20 mg L-1. The growth of C. vulgaris was significantly affected above 40 mg L-1 of diazinon, showing >30% growth inhibition after 12 days of cultivation. Significant enhancement of the microalgal growth in the exponential growth phase suggested a less/non-toxic nature of the diazinon by-products. Biochemical properties, including carotenoid, chlorophyll and antioxidant enzymes of C. vulgaris were influenced by diazinon at relatively high concentrations. The degradation rate constant (k) and the half-life (T1/2) of diazinon (0.5-100 mg L-1) ranged between 0.2304-0.049 d(-1) and 3.01-14.06 d, respectively. Gas chromatography mass spectroscopic (GC-MS) study suggested the formation of a less toxic by-product, 2-isopropyl-6-methyl-4-pyrimidinol (IMP) as a result of microalgal metabolism of diazinon. This study demonstrated that C. vulgaris is highly tolerant of diazinon, which could be voluntarily involved in the removal of traces of diazinon from contaminated wastewater and has potential application in the removal of such artificial toxins using algae. (C) 2016 Elsevier B.V. All rights reserved.</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>

      • Monitoring the gradual biodegradation of dyes in a simulated textile effluent and development of a novel triple layered fixed bed reactor using a bacterium-yeast consortium

        Kurade, M.B.,Waghmode, T.R.,Patil, S.M.,Jeon, B.H.,Govindwar, S.P. Elsevier 2017 Chemical engineering journal Vol.307 No.-

        Textile industry effluents contain a variety of dyes, which are normally resistant to biodegradation. A bacterial-yeast consortium (Brevibacillus laterosporus and Galactomyces geotrichum) was used for decolorization of two real textile effluents (RTE) and a simulated synthetic effluent (SSE). It showed enhanced decolorization compared to that of individual microorganisms with decolorization efficiency of 89, 60 and 69% for RTE-1, RTE-2 and SSE respectively, within 48h. The cumulative action of oxido-reductive enzyme in the consortium was responsible for improved decolorization. Spectroscopic analysis suggested effective biodegradation of dyes present in the SSE by the consortium contrarily to the individual strains. The gradual biodegradation of each dye present in the SSE was monitored using high performance thin layer chromatography (HPTLC). The consortium biodegraded all of the dyes within 1has compared to that of partial biodegradation by the individual microorganisms. A novel, triple layered fixed bed reactor was designed for continuous decolorization of effluent. It showed >80% decolorization (at 100mLh<SUP>-1</SUP>flow-rate), for a period of 7days, along with ~78% reduction in COD. The reproducibility of the bioreactor could be maintained for three consecutive cycles (7dayseach).

      • SCISCIESCOPUS

        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>

      • KCI등재

        Biodegradation of Disperse Dye Brown 3REL by Microbial Consortium of Galactomyces geotrichum MTCC 1360 and Bacillus sp. VUS

        S. U. Jadhav,U. U. Jadhav,V. V. Dawkar,S. P. Govindwar 한국생물공학회 2008 Biotechnology and Bioprocess Engineering Vol.13 No.2

        The consortium-GB (Galactomyces geotrichum MTCC 1360 and Bacillus sp. VUS) exhibited 100% decolorization ability with the dye Brown 3REL within 2 h at shaking condition with optima of pH 7 and at 50℃. However, G. geotrichum MTCC 1360 showed 39% decolorization within 24 h and Bacillus sp. VUS took 5 h for 100% decolorization, when incubated individually. Additional carbon and nitrogen sources like, starch, peptone, and urea were found to enhance decolorization. Induction in lignin peroxidase, tyrosinase, and riboflavin reductase was observed in consortium as that of individual organisms. GCMS identification showed different metabolites formed using consortium (2-(6,8-dichloro-quinazolin-4yloxy)-acetyl-urea and 2-(6,8-dichloro-quinazolin-4yloxy)-acetyl-formamide) and Bacillus sp. VUS (6,8-dichloro-4 methoxy-quinazoline) after 2 h of incubation with Brown 3REL. G. geotrichum MTCC 1360 showed minor modifications in structure of Brown 3REL. Phytotoxicity revealed non toxic nature of metabolites. This consortium-GB was also able to decolorize various industrial dyes.

      • KCI등재

        Purification and Characterization of Veratryl Alcohol Oxidase from Comamonas sp. UVS and Its Role in Decolorization of Textile Dyes

        Umesh U. Jadhav,Vishal V. Dawkar,Dhawal P. Tamboli,Sanjay P. Govindwar 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.3

        In the present work, we have purified veratryl alcohol oxidase (VAO) enzyme from Comamonas sp. UVS to evaluate its potential to decolorize textile dyes. VAO was purified (13.9 fold) by an ion exchange followed by the size exclusion chromatography. Molecular weight of the VAO was estimated to be about 66 kDa by SDS-PAGE. The optimum pH and temperature of oxidase were 30°C and 65°C, respectively. VAO showed maximum activity with n-propanol among the various substrates (n-propanol, veratryl alcohol, L-dopa, tryptophan, etc.). Under standard assay conditions, Km value of the enzyme was 2.5 mM towards veratrole. The enzyme activity was completely inhibited by 0.5 mM sodium azide. L-cysteine, dithiothreitol, and the metal chelator, EDTA had a slight inhibitory effect. The purified enzyme was able to decolorize textile dyes, Red HE7B (57.5%) and Direct Blue GLL (51.09%) within 15 h at 40 μg/mL concentration. GC-MS analysis of the metabolites suggested oxidative cleavage and desulphonation of these dyes In the present work, we have purified veratryl alcohol oxidase (VAO) enzyme from Comamonas sp. UVS to evaluate its potential to decolorize textile dyes. VAO was purified (13.9 fold) by an ion exchange followed by the size exclusion chromatography. Molecular weight of the VAO was estimated to be about 66 kDa by SDS-PAGE. The optimum pH and temperature of oxidase were 30°C and 65°C, respectively. VAO showed maximum activity with n-propanol among the various substrates (n-propanol, veratryl alcohol, L-dopa, tryptophan, etc.). Under standard assay conditions, Km value of the enzyme was 2.5 mM towards veratrole. The enzyme activity was completely inhibited by 0.5 mM sodium azide. L-cysteine, dithiothreitol, and the metal chelator, EDTA had a slight inhibitory effect. The purified enzyme was able to decolorize textile dyes, Red HE7B (57.5%) and Direct Blue GLL (51.09%) within 15 h at 40 μg/mL concentration. GC-MS analysis of the metabolites suggested oxidative cleavage and desulphonation of these dyes

      • SCISCIESCOPUS
      • KCI등재

        Fungal Production of Single Cell Oil Using Untreated Copra Cake and Evaluation of Its Fuel Properties for Biodiesel

        ( Mahesh Khot ),( Rohini Gupta ),( Kadambari Barve ),( Smita Zinjarde ),( Sanjay Govindwar ),( Ameeta Ravikumar ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.4

        This study evaluated the microbial conversion of coconut oil waste, a major agro-residue in tropical countries, into single cell oil (SCO) feedstock for biodiesel production. Copra cake was used as a low-cost renewable substrate without any prior chemical or enzymatic pretreatment for submerged growth of an oleaginous tropical mangrove fungus, Aspergillus terreus IBB M1. The SCO extracted from fermented biomass was converted into fatty acid methyl esters (FAMEs) by transesterification and evaluated on the basis of fatty acid profiles and key fuel properties for biodiesel. The fungus produced a biomass (8.2 g/l) yielding 257 mg/g copra cake SCO with ~98% FAMEs. The FAMEs were mainly composed of saturated methyl esters (61.2%) of medium-chain fatty acids (C12-C18) with methyl oleate (C18:1; 16.57%) and methyl linoleate (C18:2; 19.97%) making up the unsaturated content. A higher content of both saturated FAMEs and methyl oleate along with the absence of polyunsaturated FAMEs with ≥4 double bonds is expected to impart good fuel quality. This was evident from the predicted and experimentally determined key fuel properties of FAMEs (density, kinematic viscosity, iodine value, acid number, cetane number), which were in accordance with the international (ASTM D6751, EN 14214) and national (IS 15607) biodiesel standards, suggesting their suitability as a biodiesel fuel. The low cost, renewable nature, and easy availability of copra cake, its conversion into SCO without any thermochemical pretreatment, and pelleted fungal growth facilitating easier downstream processing by simple filtration make this process cost effective and environmentally favorable.

      • KCI등재

        Peroxidase from Bacillus sp. VUS and Its Role in the Decolorization of Textile Dyes

        Vishal V. Dawkar,Umesh U. Jadhav,Amar A. Telke,Sanjay P. Govindwar 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.3

        Peroxidase was purified by an ion exchange chromatography followed by gel filtration chromatography from dye degrading Bacillus sp. strain VUS. The optimum pH and temperature of the enzyme activity was 3.0 and 65°C, respectively. This enzyme showed more activity with n-propanol than other substrates tested viz. xylidine, 3-(3,4-dihydroxy phenyl) Lalanine (L-DOPA), hydroxyquinone, ethanol, indole, and veratrole. Km value of the enzyme was 0.076 mM towards n-propanol under standard assay conditions. Peroxidase was more active in presence of the metal ions like Li²+ , Co²+ , K²+ , Zn²+, and Cu²+ where as it showed less activity in the presence of Ca²+ and Mn²+ . Inhibitors like ethylenediamine tetraacetic acid (EDTA), glutamine, and phenylalanine inhibited the enzyme partially, while sodium azide (NaN3) completely. The crude as well as the purified peroxidase was able to decolourize different industrial dyes. This enzyme decolourized various textile dyes and enhanced percent decolourization in the presence of redox mediators. Aniline was the most effective redox mediator than other mediators tested. Gas chromatography-Mass spectrometry (GC-MS) confirmed the formation of 7-Acetylamino-4-hydroxy-naphthalene-2-sulphonic acid as the final product of Reactive Orange 16 indicating asymmetric cleavage of the dye Peroxidase was purified by an ion exchange chromatography followed by gel filtration chromatography from dye degrading Bacillus sp. strain VUS. The optimum pH and temperature of the enzyme activity was 3.0 and 65°C, respectively. This enzyme showed more activity with n-propanol than other substrates tested viz. xylidine, 3-(3,4-dihydroxy phenyl) Lalanine (L-DOPA), hydroxyquinone, ethanol, indole, and veratrole. Km value of the enzyme was 0.076 mM towards n-propanol under standard assay conditions. Peroxidase was more active in presence of the metal ions like Li²+ , Co²+ , K²+ , Zn²+, and Cu²+ where as it showed less activity in the presence of Ca²+ and Mn²+ . Inhibitors like ethylenediamine tetraacetic acid (EDTA), glutamine, and phenylalanine inhibited the enzyme partially, while sodium azide (NaN3) completely. The crude as well as the purified peroxidase was able to decolourize different industrial dyes. This enzyme decolourized various textile dyes and enhanced percent decolourization in the presence of redox mediators. Aniline was the most effective redox mediator than other mediators tested. Gas chromatography-Mass spectrometry (GC-MS) confirmed the formation of 7-Acetylamino-4-hydroxy-naphthalene-2-sulphonic acid as the final product of Reactive Orange 16 indicating asymmetric cleavage of the dye

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