Enzymatic analysis, structural study and molecular docking of laccase and catalase from <i>B. subtilis</i> SK1 after textile dye exposure

Kadam, Suhas K.,Tamboli, Asif S.,Sambhare, Susmit B.,Jeon, Byong-Hun,Govindwar, Sanjay P. Elsevier 2018 Ecological Informatics Vol.48 No.-

<P><B>Abstract</B></P> <P>The textile dye decolorizing efficiency of <I>Bacillus subtilis</I> SK1 against 70 mg/L each of Malachite Green, Methyl Orange, Rubine GFL and Red HE3B was observed as 71.7, 73.6, 74.4 and 82.6%, respectively within 3 h. UV–Vis spectroscopy, GC–MS and HPTLC analysis confirmed mineralization of model dyes into its metabolites. Physico-chemical characterization confirmed acidic and hydrophilic nature of both laccase and catalase enzymes. Both enzymes contain dominant random coiled secondary structure (SOPMA tool) and intracellular location (CELLO_v.2.5), however, laccase alone contains two disulfide bridges (CYS_REC tool). The validation of constructed 3D structure (Modeller 9.19) of laccase and catalase enzymes revealed, RAMPAGE- 96.3 and 95.8% residues in favoured region, ProSA- Z score −8.2 and −9.6, respectively and ERRAT-Overall quality factor > 68. Potential energies −1.328 × 10<SUP>6</SUP> kJ/mol and −2.685 × 10<SUP>6</SUP> kJ/mol remained constant after 1395 and 1545 steps for laccase and catalase, respectively in energy minimization. Molecular docking results showed interaction of Methyl Orange with laccase (Thr260) and catalase (Lys 48), Rubine GFL with laccase (Thr 262) and catalase (His 176) and Red HE3B with laccase (Asn 264, Thr 418, Gly 321, Thr 262 and Gly 378) and catalase (Gln 258). This study provides dye degrading potential of <I>Bacillus subtilis</I> strain SK1 with structurally different textile dyes and vital role of the polar amino acids of laccase and catalase in these interactions.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Bacillus subtilis</I> SK1 showed efficient remediation of different textile dyes. </LI> <LI> Laccase and catalase are accounted for degradation of structurally different dyes. </LI> <LI> Enzymes model fall in favourable region and has good quality of structure. </LI> <LI> Polar amino acids play a vital role in biotransformation of dyes. </LI> </UL> </P>

Mechanistic antimicrobial approach of extracellularly synthesized silver nanoparticles against gram positive and gram negative bacteria

Tamboli, D.P.,Lee, D.S. Elsevier Scientific Pub. Co 2013 Journal of hazardous materials Vol.260 No.-

The development of eco-friendly and reliable processes for the synthesis of nanoparticles has attracted considerable interest in nanotechnology. In this study, an extracellular enzyme system of a newly isolated microorganism, Exiguobacterium sp. KNU1, was used for the reduction of AgNO<SUB>3</SUB> solutions to silver nanoparticles (AgNPs). The extracellularly biosynthesized AgNPs were characterized by UV-vis spectroscopy, Fourier transform infra-red spectroscopy and transmission electron microscopy. The AgNPs were approximately 30nm (range 5-50nm) in size, well-dispersed and spherical. The AgNPs were evaluated for their antimicrobial effects on different gram negative and gram positive bacteria using the minimum inhibitory concentration method. Reasonable antimicrobial activity against Salmonella typhimurium, Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus was observed. The morphological changes occurred in all the microorganisms tested. In particular, E. coli exhibited DNA fragmentation after being treated with the AgNPs. Finally, the mechanism for their bactericidal activity was proposed according to the results of scanning electron microscopy and single cell gel electrophoresis.

Association between age at first calving, first lactation traits and lifetime productivity in Murrah buffaloes

Tamboli P.,Bharadwaj A.,Chaurasiya A.,Bangar Y. C.,Jerome A. 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.8

Objective: This study was conducted to estimate the association of age at first calving (AFC) with first lactation traits as well as lifetime performance traits in Murrah buffaloes. Methods: Data on first lactation and life time performance of Murrah buffaloes (n = 679), maintained at Indian Council of Agricultural Research-Central Institute for Research on Buffaloes, Hisar, India during the period 1983 through 2017, were deduced to calculate heritability estimates, genetic and phenotypic correlation of different first lactation and lifetime traits. The univariate animal model was fitted to estimate variance components and heritability separately for each trait, while bivariate animal models were set to estimate genetic and phenotypic correlations between traits under study. Results: The heritability was high for first peak milk yield (FPY, 0.64±0.08), moderate for AFC (0.48±0.07) and breeding efficiency (BE 0.39±0.09). High genetic correlations of first lactation total milk yield (FLTMY) with first lactation standard milk yield (FLSMY, 305 days or less), FPY, and first lactation length (FLL) was seen. Likewise, genetic correlation of AFC was positive with FLTMY, FLL, first dry period (FDP), first service period (FSP), first calving interval (FCI), herd life (HL) and productive days (PD). Significant phenotypic correlation of FLTMY was observed with HL, productive life (PL), PD, total lifetime milk yield (LTMY), standard lifetime milk yield (standard LTMY). Moreover, positive genetic and phenotypic correlation of FPY was observed with HL, PL, PD, total LTMY and standard LTMY. Conclusion: This study reports that AFC had positive genetic correlation with FDP, FSP, FCI, and unproductive days while, negative association of AFC was observed with FLSMY, PL, total LTMY, standard LTMY, and BE. This suggests that reduction of AFC would results in improvement of lifetime performance traits. Objective: This study was conducted to estimate the association of age at first calving (AFC) with first lactation traits as well as lifetime performance traits in Murrah buffaloes.Methods: Data on first lactation and life time performance of Murrah buffaloes (n = 679), maintained at Indian Council of Agricultural Research-Central Institute for Research on Buffaloes, Hisar, India during the period 1983 through 2017, were deduced to calculate heritability estimates, genetic and phenotypic correlation of different first lactation and lifetime traits. The univariate animal model was fitted to estimate variance components and heritability separately for each trait, while bivariate animal models were set to estimate genetic and phenotypic correlations between traits under study.Results: The heritability was high for first peak milk yield (FPY, 0.64±0.08), moderate for AFC (0.48±0.07) and breeding efficiency (BE 0.39±0.09). High genetic correlations of first lactation total milk yield (FLTMY) with first lactation standard milk yield (FLSMY, 305 days or less), FPY, and first lactation length (FLL) was seen. Likewise, genetic correlation of AFC was positive with FLTMY, FLL, first dry period (FDP), first service period (FSP), first calving interval (FCI), herd life (HL) and productive days (PD). Significant phenotypic correlation of FLTMY was observed with HL, productive life (PL), PD, total lifetime milk yield (LTMY), standard lifetime milk yield (standard LTMY). Moreover, positive genetic and phenotypic correlation of FPY was observed with HL, PL, PD, total LTMY and standard LTMY.Conclusion: This study reports that AFC had positive genetic correlation with FDP, FSP, FCI, and unproductive days while, negative association of AFC was observed with FLSMY, PL, total LTMY, standard LTMY, and BE. This suggests that reduction of AFC would results in improvement of lifetime performance traits.

Purification and Characterization of Bacterial Aryl Alcohol Oxidase from Sphingobacterium sp. ATM and Its Uses in Textile Dye Decolorization

Dhawal P. Tamboli,Amar A. Telke,Vishal V. Dawkar,Shekhar B. Jadhav,Sanjay P. Govindwar 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.4

Aryl alcohol oxidase (AAO) produced by dye decolorizing bacteria Sphingobacterium sp. ATM, was purified 22.63 fold to a specific activity of 21.75 μmol/min/mg protein using anion exchange and size exclusion chromatography. The molecular weight of the purified AAO was found to be 71 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE),and confirmed by zymography of AAO using L-dopa. The enzyme showed substrate specificity towards veratryl alcohol, followed by n-propanol. The optimum pH and temperature of purified AAO were found to be 3.0 and 40°C, respectively. The K_m and V_(max) of AAO was 1.1615mM and 3.13 mM/min when veratryl alcohol was used as substrate. Sodium azide showed maximum inhibition while ethylenediamine tetra acetic acid (EDTA), L-cysteine and dithiothreitol showed slight inhibition. Metal ions also showed slight inhibition. HPLC analysis confirmed the degradation of Direct Red 5B. The metabolite obtained after decolorization of Direct Red 5B was characterized as 3 diazenyl 7 [-(phenyl carbonyl) amino] naphthalene-2-sulfonic acid using GC-MS analysis.

Little known Curculigo brevifolia deserves a species rank: evidences from morphological, cytological and molecular data

Avinash R. Gholave,Asif S. Tamboli,Rohit N. Mane,Ramchandra D. Gore,박재홍,Sayajirao P. Gaikwad 국립중앙과학관 2021 Journal of Asia-Pacific Biodiversity Vol.14 No.4

The Curculigo brevifolia resurrected here based on morphological, cytological, and molecular phylogeneticanalyses. Curculigo brevifolia morphologically resembles C. orchioides but differs in having bulbils atthe tip of leaves for vegetative reproduction, leaves elongated falcately shaped, beaked fruits. Cytologicalparameters, karyotype formula, total haploid genome length (THL), values of CVCL, MCA also supportedthe distinctness of C. brevifolia and C. orchioides. Phylogenetic analysis based on cpDNA data resulted inthe recognition of three clades and strongly supported the revised systematics of Hypoxidaceae. Theresurrected species nests within the Curculigo clade of Hypoxidaceae and display a close phylogeneticaffinity with newly added Curculigo species and C. orchioides.

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

Degradation and Detoxification of Disperse Dye Scarlet RR by Galactomyces geotrichum MTCC 1360

S. U. Jadhav,G. S. Ghodake,A. A. Telke,D. P. Tamboli,S. P. Govindwar 한국미생물 · 생명공학회 2009 Journal of microbiology and biotechnology Vol.19 No.4