International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 36 Use of Biological Database to Explore Microorganisms used in Bioremediation of Hexavalent Chromium Hajoori M.A.and Tank S.K. Department of Biosciences, Veer Narmad South Gujarat University, Surat, INDIAAvailable online at: www.isca.in, www.isca.me Received 11th Septmeber 2015, revised 28th September 2015, accepted 5th October 2015 Abstract Bioinformatics is one of the newest emerging fields of life sciences. Number of biological database has been created to provide vital information regarding module of interest. The present study was aimed to utilize these biological databases to explore significant microorganisms that had got an ability to reduce hexavalent chromium. Using Text search tool available at Protein Information Resources database, total 906 entries were obtained, out of which 49 entries were filtered depicting different microorganisms having chromate reductase enzyme. Five microorganisms belonging to Pseudomonas, Bacillus, Geobacillus, Arthrobacter and Staphylococcus species show positive screening for reduction of hexavalent chromium. Thus, use of biological database provides an aid to explore the microorganisms having potential in bioremediation of chromium compounds. The bacterial isolates having ability to convert toxic form of chromium into their nontoxic form can be employed for bioremediation of hexavalent chromium. Keywords: Hexavalent chromium, biological database, bioremediation, protein information resources. Introduction A large numbers of toxic chemicals (pollutants) are generated from various industries, releasing it into surrounding environment, where they contaminate soil and water1-2. Some of these toxic chemicals lead to cause severe problem to living organisms and required to be removed from environment. Microorganisms due to their versatile nutritional requirement can be used to degrade such toxic compound4-6. Chromium compounds are one of such toxic chemicals generated from various industries like tannery, textile, steel, paint and galvanization units. The chromium exists in nature primarily as highly toxic hexavalent chromium and least toxic trivalent chromium8-9. Chromium has its carcinogenic10-11 and mutagenicity12-13 effects. Hence, ranked 18th in priority list of hazardous substances issued by CERCLA, 200714. USEPA has categories chromium as class a pollutant15. Various mechanisms were known for bioremediation of hexavalent compounds. One of such mechanism includes bioremediation by a group of enzyme chromate reductases16. Numbers of biological databases has been available consisting of number of information pertaining to molecule of interest. Hereby attempt has been made in this study to use information from biological database to incur potential microorganisms that can show effective bioconversion of hexavalent chromium into trivalent non toxic form. Material and Methods Exploring microorganisms with chromate reductase enzyme: Microorganisms having enzyme chromate reductase were explored using text search tools available at Protein information resources database17. The available list of microorganisms was further filtered to remove redundant entries having same genus and species. Selection of microorganisms was carried out on the basis of their availability from natural environment or culture collection centre and their pathogenic nature. Pair wise sequence alignment: The pair wise sequence alignments were performed using pair wise alignment mode available at protein information resources18 for selected entries to determine the similarity and dissimilarity between the chromate reductase which may contribute to variation in their activity to reduce hexavalent chromium into their non-toxic product. Isolation of selected microorganisms: The microorganisms viz., Pseudomonas spp., Bacillus spp. and Staphylococcus spp. identified having chromate reductase enzyme were isolated from soil samples collected from vicinities of dyes and chemical industries near Palsana, Surat, Gujarat, India. The soil sample was mixed with sterile distilled water and aliquots of 10-1, 10-2, 10-3, 10-4, 10-5 and 10-6 were prepared. Each dilution was plated onto Nutrient agar plate and incubated at 37°C for 24 Hr. Geobacillus spp was procured from HImedia whereas Arthrobacter was procured from culture collection centre. Pure culture of all the strain was prepared using nutrient agar medium. Screening of microorganisms for chromium reduction:The isolate were precultured overnight on nutrient agar medium. culture flask was prepared with basal medium (Glucose 0.5 g, Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 37 calcium chloride 0.02 g, monopotassium phosphate 1.0 g, ferric chloride 0.05 g, dipotassium phosphate 1.0 g, magnesium sulphate 0.2 g, ammonium nitrate 1.0 g and distilled water 1000 ml) supplemented with KCr having chromium (VI) concentration of 100 ppm. 3.5 ml of isolated culture was inoculated into 350 ml of basal medium with KCr. The inoculated flasks were incubated at room temperature on rotary shaker having speed of 150 RPM. Samples were drawn at an interval of 0 hr, 12 hr, 24 hr, 36 hr and 48 hr to determine growth of isolate measured in terms of optical density at 540 nm using a SHIMADZU UV-Spectrophotometer against uninoculated basal medium containing chromium as blank. Determination of Hexavalent Chromium: The supernatant of each flask after centrifugation were analyzed for Chromium (VI) using 1, 5-Diphenyl Carbazide method as described by USEPA19. Chromium Standard with chromium concentration ranges from 100 ppm to 1000 ppm was prepared. 95 ml of the extract were centrifuge and 2.0 ml diphenylcarbazide solution was added. pH of 2 + 0.5 was set with 10% HSO solution and dilute to final volume of 100 ml. Colour developed was estimated spectrophotometrically at 540 nm using Basal medium as Blank. Protein 3D Structure Prediction: Chromate reductase protein sequence of selected microorganisms was used for protein structure modeling using Swiss-Model20-23 server available online at server of Swiss Institute of Bioinformatics. The structures were building using automated mode that identifies best templates based on Blast24 and HHblits25. Multiple Structure Alignment: All the modeled protein structures were then superimposed using Mistral tool26 for multiple structure alignment of protein to evaluate the similarity and dissimilarity with respect to protein structure. This help to identify the conserved and non-conserved region that may contribute for variation in chromate reductase activity. The Mistral tool performed multiple structure alignment of protein on the basis of energy minimization and translations of the given molecule. Results and Discussion Exploring microorganisms having Chromate Reductase:Using text search tools available at Protein Information Resources database, total 906 entries were obtained which was subjected to further screening for list of microorganisms possessing this enzyme. Further, filtration of obtained list give total 49 different microorganisms table-1 having chromate reductase. 5 different microorganisms table-2 were selected from the list of 49 microorganisms. Figure-1 Pairwise Sequence Alignment [Pseudomonas spp. Vs. Bacillus spp.] Research Journal of Biological Sciences _ _____________________ Vol. 4(10), 36-47, October (2015) International Science Congress Association List of mic Protein AC/ID P0AGE6/CHRR_ECOLI Chromate reductase P0AGE8/CHRR_SHIFL Chromate reductase Q88FF8/CHRR_PSEPK Chromate reductase A0A031QPU8/A0A031QPU8_SERMA Chromate reductase, Class I, flavoprotein A0A060VC33/A0A060VC33_KLESP Chromate reductase, Class I, flavoprotein A1K9U9/A1K9U9_AZOSB A3I992/A3I992_9BACI Chromate reductase A6EA35/A6EA35_9SPHI B0JDW3/B0JDW3_THESC Chromate reductase B3VBK2/B3VBK2_ARTAU Chromate reductase B3VBK3/B3VBK3_BACAT Chromate reductase B4F0I4/B4F0I4_PROMH reductase (NADPH G8MY11/G8MY11_GEOTH Chromate reductase I5BT83/I5BT83_9RHIZ Chromate reductase I7MR59/I7MR59_STRCB J0MD04/J0MD04_9ENTR Chromate reductase J1GP88/J1GP88_9LACT Chromate reductase J7LWV5/J7LWV5_9MICC J9YFA6/J9YFA6_LEUGJ Chromate reductase K2MJB1/K2MJB1_9RHIZ Chromate reductase K2QFK6/K2QFK6_9RHIZ Chromate reductase K5ZL66/K5ZL66_9PROT Chromate reductase L7ZF37/L7ZF37_SERMA Chromate reductase, Class I, flavoprotein M1GF39/M1GF39_MYCPM _____________________ _________________________ ____________ International Science Congress Association Table-1 List of mic roorganisms having chromate reductase Protein Name Length Organism Name Chromate reductase 188 Escherichia coli (strain K12) Chromate reductase 188 Shigella flexneri Chromate reductase 186 Pseudomonas putida (strain KT2440) Chromate reductase, Class I, flavoprotein 188 Serratia marcescens BIDMC 81 Chromate reductase, Class I, flavoprotein 188 Klebsiella sp Probable chromate reductase 186 Azoarcus sp. (strain BH72) Chromate reductase 180 Bacillus sp. B14905 Putative chromate reductase 185 Pedobacter sp. BAL39 Chromate reductase 349 Thermus scotoductus Chromate reductase (Fragment) 75 Arthrobacter aurescens Chromate reductase (Fragment) 75 Bacillus atrophaeus Putative chromate reductase (NADPH dependent FMN reductase) 182 Proteus mirabilis (strain HI4320) Chromate reductase 181 Geobacillus thermoleovorans CCB_US3_UF5 Chromate reductase 186 Nitratireductor aquibiodomus RA22 Putative chromate reductase 146 Streptococcus canis FSL Z3-227 Chromate reductase monomer 188 Enterobacter sp. Ag1 Chromate reductase 179 Weissella koreensis KCTC 3621 Putative chromate reductase 203 Arthrobacter sp. Rue61a Chromate reductase 181 Leuconostoc gelidum (strain JB7) Chromate reductase 186 Nitratireductor pacificus pht-3B Chromate reductase 175 Agrobacterium albertimagni AOL15 Chromate reductase 180 Acidocella sp. MX-AZ02 Chromate reductase, Class I, flavoprotein 188 Serratia marcescens WW4 Putative chromate reductase 166 Mycoplasma pneumoniae M129-B7 ____________ ISSN 2278-3202 Int. Res. J. Biological Sci. 38 UniRef50 UniRef50_P0AGE7 UniRef50_P0AGE7 UniRef50_Q93T20 UniRef50_V6A4U4 UniRef50_P0AGE7 UniRef50_V6A4U4 UniRef50_D3ECI3 UniRef50_F5YA92 UniRef50_A8HNC2 UniRef50_P0AGE7 UniRef50_P0AGE7 UniRef50_A1VMH5 UniRef50_G8MY11 UniRef50_I5BT83 UniRef50_I7MR59 UniRef50_V6A4U4 UniRef50_D3ECI3 UniRef50_A0A059MSD7 UniRef50_M4KEA3 UniRef50_S5YVM0 UniRef50_K2QFK6 UniRef50_K5ZL66 UniRef50_V6A4U4 UniRef50_P47584 Research Journal of Biological Sciences _ _____________________ Vol. 4(10), 36-47, October (2015) International Science Congress Association Protein AC/ID Q08VE8/Q08VE8_STIAD Q38W12/Q38W12_LACSS Q67S97/Q67S97_SYMTH Q7VY03/Q7VY03_BORPE Q8CUR9/Q8CUR9_OCEIH Chromate reductase R4Y2X0/R4Y2X0_ALCXX S5U1V3/S5U1V3_PROMI Chromate (NADPH S6CM53/S6CM53_9PROT Chromate reductase S7U7F5/S7U7F5_9BACI Chromate reductase T1Y622/T1Y622_STAAU Chromate reductase U2Y9E7/U2Y9E7_GEOKU Chromate reductase V6MI66/V6MI66_PROHU Chromate reductase V7EMG5/V7EMG5_9RHOB Chromate reductase W0XIT2/W0XIT2_KLEPN Chromate reductase, Class I, flavoprotein W1SPF4/W1SPF4_9BACI Chromate reductase W4N9W7/W4N9W7_9BIFI reductase/NADPH W4Q5N1/W4Q5N1_9BACI W5X189/W5X189_BDEBC Chromate reductase, Class I, flavoprotein W5Y707/W5Y707_KOMXY Chromate reductase _____________________ _________________________ ____________ International Science Congress Association Protein Name Length Organism Name Putative chromate reductase 291 Stigmatella aurantiaca (strain DW4/3-1) Putative chromate reductase 185 Lactobacillus sakei subsp. sakei (strain 23K) Putative chromate reductase 335 Symbiobacterium thermophilum (strain T / IAM 14863) Putative chromate reductase 184 Bordetella pertussis (strain Tohama I / ATCC BAA-589 / NCTC 13251) Chromate reductase 181 Oceanobacillus iheyensis (strain DSM 14371 / JCM 11309 / KCTC 3954 / HTE831) Putative chromate reductase 184 Achromobacter xylosoxidans NH44784-1996 Chromate reductase (NADPH -dependent FMN reductase) 182 Proteus mirabilis BB2000 Chromate reductase (Fragment) 103 proteobacterium S3K Chromate reductase 272 Geobacillus sp. WSUCF1 Chromate reductase 268 Staphylococcus aureus subsp. aureus CN1 Chromate reductase 180 Geobacillus kaustophilus GBlys Chromate reductase 182 Proteus hauseri ZMd44 Chromate reductase 178 Rhodobacter sp. CACIA14H1 Chromate reductase, Class I, flavoprotein 188 Klebsiella pneumoniae subsp. pneumoniae T69 Chromate reductase 184 Bacillus vireti LMG 21834 Chromate reductase/NADPH dependent FMN reductase/Oxygen-insensitive NA... 262 Bifidobacterium moukalabense DSM 27321 Putative chromate reductase 181 Bacillus wakoensis JCM 9140 Chromate reductase, Class I, flavoprotein 183 Bdellovibrio bacteriovorus W Chromate reductase 186 Gluconacetobacter xylinus E25 ____________ ISSN 2278-3202 Int. Res. J. Biological Sci. 39 UniRef50 UniRef50_F8CBU6 UniRef50_Q93T20 UniRef50_Q67S97 UniRef50_A1VMH5 UniRef50_D3ECI3 UniRef50_A1VMH5 UniRef50_A1VMH5 UniRef50_P0AGE7 UniRef50_P39605 UniRef50_Q8CMQ4 UniRef50_G8MY11 UniRef50_A1VMH5 UniRef50_V7EMG5 UniRef50_P0AGE7 UniRef50_G8MY11 UniRef50_R5NFV9 UniRef50_R9X5A0 UniRef50_Q93T20 UniRef50_V6A4U4 Research Journal of Biological Sciences _ _____________________ Vol. 4(10), 36-47, October (2015) International Science Congress Association Protein AC/ID W6EP16/W6EP16_BIFBR reductase/NADPH W8FZ94/W8FZ94_9GAMM Chromate reductase W8X1A1/W8X1A1_BIFAN Chromate reductase X5IHB9/X5IHB9_BORBO Chromate re X7EE43/X7EE43_9RHOB Chromate reductase UPI00029C4B42_587 chromate reductase UPI0003900F2B_562 chromate reductase, Class I, flavoprotein Pair wise Sequence _____________________ _________________________ ____________ International Science Congress Association Protein Name Length Organism Name Chromate reductase/NADPH dependent FMN reductase/Oxygen-insensitive NA... 255 Bifidobacterium breve 12L Chromate reductase 176 Thalassolituus oleivorans R6-15 Chromate reductase 376 Bifidobacterium animalis subsp. lactis CECT 8145 Chromate re ductase 184 Bordetella bronchiseptica (Alcaligenes bronchisepticus) Chromate reductase 189 Roseivivax halodurans JCM 10272 chromate reductase 185 Providencia rettgeri chromate reductase, Class I, flavoprotein 176 Escherichia coli Figure-2 wise Sequence Alignment [ Pseudomonas spp. Vs. Arthrobacter spp. ____________ ISSN 2278-3202 Int. Res. J. Biological Sci. 40 UniRef50 UniRef50_R5NFV9 UniRef50_W8FZ94 UniRef50_E4R026 UniRef50_A9HXL5 UniRef50_X7EE43 UniRef50_A1VMH5 UniRef50_P0AGE7 Pseudomonas spp. Vs. Arthrobacter spp. ] Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 41 Figure-3 Pair wise Sequence Alignment [Pseudomonas spp. Vs. Geobacillus spp.] Figure-4 Pair wise Sequence Alignment [Pseudomonas spp. Vs. Staphylococcus spp.] Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 42 Isolation and screening and characterization of selected microorganisms: 34 different isolates were obtained from collected sample and 5 screened for survival in Basal medium containing KCr. All the five strain were evaluated for tolerance in term of growth figure-5. The cell mass of Pseudomonas spp. show highest growth in the medium supplemented with chromium after 36 and 48 hours27. The acillus spp. and Arthrobacter spp. show increase in growth after 36 hours of incubation28. Staphylococcus spp. show least growth pattern suggest less tolerance toward chromium29. The Geobacillus show increase in growth after 36 hr of incubation. Thus, these microorganisms possess ability to tolerate hexavalent chromium at the concentration of 100 ppm. However difference in sequence may contribute for variation in their ability to reduce hexavalent chromium. The results obtained suggest all selected microorganisms had ability for reducing hexavalent chromium. The results were in accordance with different authors7, 27-29. The selected isolates were further evaluated for their activity for reduction of hexavalent chromium. Standard for chromium was prepared using concentration from 100 to 1000ppm. Determination of Isolate ability to reduce hexavalent chromium: All the five isolate were evaluate for reduction in hexavalent chromium. Pseudomonas species show 42.3%, Bacillus species show 34.5%, Arthrobacter species show 29.8%, Staphylococcus spp show 28% and Geobacillus about 31.6% reduction of hexavalent chromium after 48 hrs of incubation with chromium concentration of 100 ppm figure-6. Figure-5 Biomass determination of selected isolates Figure-6 Activity of Isolate for Hexavalent chromium reduction 0.050.10.150.20.250.312 hr24 hr36 hr48 hrO.D.Time Pseduomonas spp. Bacillus Spp Staphylococcus spp. Geobacillus spp. Arthrobacter spp. 1015202530354045 % Reduction (Cr-VI)Isolates % reduction Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 43 Protein 3D Structure Prediction: All the Structure prediction of chromate reductase were conducted using Swiss model template library search using Blast24 and HHBlits25 to determine evolutionary relatedness of the structures matching the target sequence of chromate reductase available in database. The templates found by Swiss model was overall 764 templates for Pseudomonas spp, 492 templates for Bacillus spp.,146 templates for Staphylococcus spp., 667 for Arthrobacter spp. and 127 templates for Geobacillus spp. Out of these entire template 03 models from each category was build on the basis of identity score. The best model from each category was selected on the basis of global and preresidual model quality expressed in terms of QMean4 value. For Pseudomonas spp., QMean4 value of -4.67 with sequence identity of 49.72% and coverage of 95% were obtained figure-7. For Bacillus spp., QMean4 value of -5.44 with sequence identity of 38.42% and coverage of 98% were obtained figure-8. For Staphylococcus spp., QMean4 value of -1.50 with sequence identity of 41.13% and coverage of 93% were obtained figure-9. For Athrobacter, QMean4 value of -4.14 with sequence identity of 44.32% and coverage of 87% were obtained figure-10. For Geobacillus spp, QMean4 value of -1.50 with sequence identity of 59.51% and coverage of 91% were obtained figure-11. Figure-7 Chromate reductase 3D structure prediction of Pseudomonas spp.Figure-8 Chromate reductase 3D structure prediction of Bacillus spp. Figure-9 Chromate reductase 3D structure prediction of Arthrobacter spp. Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 44 Figure-10 Chromate reductase 3D structure prediction of Geobacillus spp. Figure-11 Chromate reductase 3D structure prediction of Staphylococcus spp.Chromate reductase modeled of Pseudomonas, Arthrobacterand Bacillus was building using same template 3u7r.1.A as reference. Thus, all model predicted of Pseudomonas, Arthrobacter and Bacillus show similarity with NADPH-dependent FMN reductase whereas chromate reductase modeled of Staphylococcus and Geobacillus was build using 3n2s.1.A and 3n2s.1.B as reference. Thus, the model predicted show similarity with NADPH-dependent nitro/flavin reductase. Multiple Structure Alignment: Mistral Software used for multiple structure alignment of all the modeled structure reveal that Pseudomonas, Arthrobacter and Bacillus species show majority of superimposition with minimum RMSD and least energy level. The best alignment was obtained showing good spatial arrangement and stability after superimposition26. Modeled chromate reductase of Geobacillus spp. and Staphylococcus spp. has higher RMSD and energy as compared to other above three chromate reductase table-3. Conclusion All the isolate show activity for chromium reduction contributed due to presence of chromate reductase enzyme. The efficiency to catalyze reaction varies due to variation in amino acid composition of chromate reductase. Using various tools and software of bioinformatics significant information can be obtained that aids in exploring microorganisms having ability to degrade such pollutants. Thus there is a need to develop specialized database showing potential candidates for waste cleanup. AcknowledgementWe are thankful to Bhagwan Mahavir College of M.Sc. Biotechnology, Surat for providing laboratory facilities. References 1.Ellis L.B. and Wackett L.P., Use of the University of Minnesota Biocatalysis/Biodegradation Database for study of microbial degradation, Microb. Inform. Exp., 2(1), 1-10 (2012)2.Arora P. and Shi W., Tools of bioinformatics in biodegradation, Rev. Environ. Sci. Biotechnol., , 211-213 (2010)3.Andrady A.L., Biodegradation of plastics: monitoring what happens, Plastics Additives. Springer Netherlands, 1, 32-40 (1998) Research Journal of Biological Sciences ___________________________________________________________ ISSN 2278-3202 Vol. 4(10), 36-47, October (2015) Int. Res. J. Biological Sci. International Science Congress Association 45 Table-2 Amino Acid Sequence of five selected microorganisms�sp|Q88FF8|CHRR_PSEPK Chromate reductase OS=Pseudomonas putida (strain KT2440) GN=PP_4138 PE=1 SV=1 MSQVYSVAVVVGSLRKESYNRKVARALSELAPSSLALKIVEIGDLPLYNEDIEAEAPPETWKRFRDEIRRSDAVLFVTPEYNRSVPGCLKNAIDVGSRPYGQSAWSGKPTAVVSVSPGAIGGFGANHAVRQSLVFLDMPCMQMPEAYLGGAASLFEDSGKLNDKTRPFLQAFVDRFASWVKLNRAV �tr|A3I992|A3I992_9BACI Chromate reductase OS=Bacillus sp. B14905 GN=BB14905_07074 PE=4 SV=1MKVVAIVGSIRKESYNMQLAQFIEKRYTEKLDLEVLSLKDLPMYNQDIENEAPQAVLDFKAKVKAADAVLWVTPEYNGTVPGVMINAIDWLSRVDKVMIGKPSIIMGASMGNLGTVKAQLHLRDILFSPGINSPLLSGNDVYIGAVHTKFDAEGNLTDEGTVKFLDVVIDNFLNWAKKYI �tr|J7LWV5|J7LWV5_9MICC Putative chromate reductase OS=Arthrobacter sp. Rue61a GN=ARUE_c41610 PE=4 SV=1 MDTFKIGYFVGSLASNSINRVLSKALISVAPPELEFHEIAIKDLPLYSADYDADFPPAGRELKDAIAASDGILFVSPEYNRSIPGALKNAIDWGSRPWGTNSFARKPTGIIGASPGGIGTAVMQSSMRSVLSFLDAPQLNAPEAYIRFVADAYDDDGSVKDEGTAGLLRHYMEEYSAFVQRVLAANAPGHIGDPEPDSAKLTR �tr|S7U7F5|S7U7F5_9BACI Chromate reductase OS=Geobacillus sp. WSUCF1 GN=I656_00399 PE=4 SV=1 MALSFLLSCSQKEAVGLFKEDIVMNQVIETILQHRSIRRFEDQPLTDEQIRTIVECAQAASTSSYVQAYSIIGVKDPEKKRKLAELAGNQSYVEHNGHFFVFCADFHRHELIGELEGKDVLPSLESTEKFMVALIDTALAAQNAAIAAESMGLGICYIGGLRNNLPEVCALLNVPKRVIPLFGLAVGYPAQTPDQKPRLPFEHVYHEDEYDQDRARFIAQLQRYNETVSTYYEQRTNGRRRDTWTGQMADMLSRQVRMYMKEFVEGKGFNLR �tr|T1Y622|T1Y622_STAAU Chromate reductase OS=Staphylococcus aureus subsp. aureus CN1 GN=SAKOR_00381 PE=4 SV=1 MQYEDFKLIVKYRGVGNVSEHVYNLVKKHHSVRKFKNKPLSEDVVKKLVEAGQSASTSSFLQAYSIIGIDDEKIKENLREVSGQPYVVENGYLFVFVIDYYRHHLVDQHAETDMENAYGSTEGLLVGAIDAALVAENIAVTAEDMGYGIVFLGSLRNDVERVREILDLPDYVFPVFGMAVGEPADDENGAAKPRLPFDHVFHHNKYHADKETQYAQMADYDQTISEYYDQRTNGNRKETWSQQIEMFLGNKARLDMLEQLQKSGLIQR 4.Arora P.K., Sasikala C. and Ramana C.V., Degradation of chlorinated nitroaromatic compounds, Appl. Microbiol. Biotechnol.,93(6), 2265-2277 (2012)5.Arora P.K., Srivastava A. and Singh V.P., Bacterial degradation of nitrophenols and their derivatives, J. Hazard. 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International Science Congress Association 46 Table-3 Multiple Structure Alignment of chromate reductase of selected microorganisms Pair: Lengths Energy N_matches RMSD sid Z Score P value model_Arthrobacter.pdb model_Bacillus.pdb 352 358 1286.605 348 3.3 35 12.8 4.17e-08 model_Arthrobacter.pdb model_Geobacillus.pdb 352 492 -361.123 78 2.8 7 1.0 1.52e-01 model_Arthrobacter.pdb model_Pseudomonas.pdb 352 356 -296.951 348 1.9 122 13.0 3.22e-08 model_Arthrobacter.pdb model_Staphyococcus.pdb 352 497 -352.923 43 3.0 3 0.9 1.68e-01 model_Bacillus.pdb model_Geobacillus.pdb 358 492 -356.979 51 2.6 3 0.9 1.59e-01 model_Bacillus.pdb model_Pseudomonas.pdb 358 356 1290.714 345 2.6 72 12.8 3.91e-08 model_Bacillus.pdb model_Staphyococcus.pdb 358 497 -367.135 71 2.7 5 1.0 1.45e-01 model_Geobacillus.pdb model_Pseudomonas.pdb 492 356 -405.222 101 2.8 4 1.4 9.40e-02 model_Geobacillus.pdb model_Staphyococcus.pdb 492 497 -1769.578 489 0.7 208 13.6 1.45e-08 model_Pseudomonas.pdb model_Staphyococcus.pdb 356 497 -378.962 78 2.7 8 1.1 1.27e-0 12.O'Brien J.C., and Patierno S.R., Effects of glutathione on chromium-induced DNA crosslinking and DNA polymerase arrest, Mole. 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Sciences., 1(4), 18-24 (2014)28.Elangovan R., Abhipsa S., Rohit B., Ligy P. amd Chandraraj K., Reduction of Cr(VI) by a Bacillus sp., Biotechnology Letters., 28, 247–252 (2006)29.Mistry K, Desai C., Lal S., Patel K. and Patel B., Hexavalent Chromium Reduction by Staphylococcus Sp. Isolated From Cr (Vi) Contaminated Land Fill, Inter. Jour. of Biotech. and Biochem.,6(1), 117–129 (2010)