Community Synergism: Degradation of Triazine Dye Reactive Black 1 by Mixed Bacterial Cultures KND_PR under Microaerophilic and Aerobic Conditions

  • Kshama Balapure
  • Payal Aghera
  • Nikhil BhattEmail author
  • Datta Madamwar
Original Article


Mixed bacterial cultures KND_PR comprising five bacterial species, namely Lysinibacillus sp. BAB-4931, Raoultella sp. BAB-4932, Enterococcus sp. BAB-4933, Citrobacter sp. BAB-4934 and Lysinibacillus sp. BAB-4935, were originally developed from contaminated soil near Pirana, Ahmedabad, India. KND_PR have an astonishing ability to degrade Reactive Black 1 (RB1) under both microaerophilic and aerobic conditions within 8 and 12 h, respectively. Mixed bacterial cultures KND_PR showed azoaromatic degradation efficiency up to 1500 mg/L of dye concentration and were able to tolerate salt up to 25 g/L. KND_PR can degrade twenty-five structurally different complex dyes that indicate its catabolic versatility. During azoaromatic degradation, it was observed that under microaerophilic condition KND_PR first followed symmetric cleavage of the azo bond by azoreductase, whereas, under aerobic condition, asymmetric cleavage of RB1 took place via lignin peroxidase. Metabolic pathways for RB1 degradation by KND_PR has been postulated with enzymatic and bioanalytical techniques, which showed the significant conversion of parent dye into low molecular weight aliphatic compound. These results were further confirmed by observing significant results of phytotoxicity experiments with model plants under microaerophilic and aerobic conditions. The foregoing results indicate that the enriched mixed bacterial cultures can efficiently degrade dye under microaerophilic and aerobic conditions within a short time period, which exhibited the usefulness of mixed bacterial cultures KND_PR to tackle the problems associated with dye contaminated wastewater.


KND_PR Enzymes Aromatics Detoxification Metabolic pathway 



The authors are thankful to Gujarat State Biotechnology Mission (GSBTM), Gandhinagar, Gujarat, India for providing research grant. Authors express their deep gratitude to Dr. Jatin Upadhyay, M.V.M Science and Home Science College, Rajkot, Gujarat, India for their valuable help in pathway elucidation. Authors also are grateful to SICART (Sophisticated Instrumentation Centre of Applied Research and Training) Vallabh Vidyanagar, Gujarat, India.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

40710_2019_378_MOESM1_ESM.docx (104 kb)
ESM 1 (DOCX 103 kb)


  1. Balapure KH, Jain K, Chattaraj S, Bhatt NS, Madamwar D (2014) Co-metabolic degradation of diazo dye - reactive blue 160 by enriched mixed cultures BDN. J Hazard Mater 279:95–95CrossRefGoogle Scholar
  2. Balapure K, Bhatt N, Madamwar D (2015) Mineralization of reactive azo dyes present in simulated textile wastewater using down flow microaerophilic fixed film bioreactor. Bioresour Technol 175:1–7CrossRefGoogle Scholar
  3. Balapure K, Jain K, Bhatt N, Madamwar D (2016) Exploring bioremediation strategies to enhance the mineralization of textile industrial wastewater through sequential anaerobic-microaerophilic process. Int Biodeterior Biodegrad 106:97–105CrossRefGoogle Scholar
  4. Bedekar P, Saratale RG, Saratale GD, Govindwar SP (2014) Development of low cost upflow column bioreactor for degradation and detoxification of blue HERD and textile effluent by Lysinibacillus sp. RGS immobilized on loofa. Int Biodeterior Biodegrad 96:112–120CrossRefGoogle Scholar
  5. Bhosale S, Saratale G, Govindwar S (2006) Biotransformation enzymes in Cunninghamella blakesleeana (NCIM-687). J Basic Microbiol 46:444–448CrossRefGoogle Scholar
  6. Chang JS, Chou G, Lin YC, Lin PJ, Ho JY, Ho TL (2001) Kinetic characteristic of bacterial azo-dye decolorization by Pseudomonas luteola. Water Res 35:2841–2850CrossRefGoogle Scholar
  7. Chanwala J, Kaushik G, Mohd Ashraf D, Upadhyay S, Agrawal A (2019) Process optimization and enhanced decolorization of textile effluent by Planococcus sp. isolated from textile sludge. Environ Technol Innov 13:122–129CrossRefGoogle Scholar
  8. Chattaraj S, Johnson J, Madamwar D (2015) Biotransformation of mixture of dyes by enriched bacterial consortium ASD. Desalin Water Treat 1:13Google Scholar
  9. Chaudhari AU, Tapase SR, Markad VL, Kodam KM (2013) Simultaneous decolorization of reactive orange M2R dye and reduction of chromate by Lysinibacillus sp. KMK-A. J Hazard Mater 262:580–588CrossRefGoogle Scholar
  10. Chen K, Wua J, Liou D, Hwang S (2003) Decolorization of the textile dyes by newly isolated bacterial strains. J Biotechnol 101:57–68CrossRefGoogle Scholar
  11. Desai CK, Pathak H, Madamwar D (2010) Advances in molecular and “-omics” technologies to gauge microbial communities and bioremediation at xenobiotic/anthropogen contaminated sites. Bioresour Technol 101:1558–1569CrossRefGoogle Scholar
  12. El Fantroussi A, Said N, Spiros D, Pieper H, Witzig R, Cámara B (2006) Biological assessment and remediation of contaminated sediments. Springer, Dordrecht, pp 179–238CrossRefGoogle Scholar
  13. Forss J, Lindh MV, Pinhassi J, Welander U (2017) Microbial biotreatment of actual textile wastewater in a continuous sequential rice husk biofilter and the microbial community involved. PLoS One 12:1–16CrossRefGoogle Scholar
  14. Franciscon E, Grossman MJ, Rizzato JA, Paschoal JAR, Reyes FA, Durrant LR (2012) Decolorization and biodegradation of reactive sulfonated azo dyes by a newly isolated Brevibacterium sp. strain VN-15. SpringerPlus 1(1):37CrossRefGoogle Scholar
  15. Hakimelahi M, Moghaddam MRA, Hashemi SH (2012) Biological treatment of wastewater containing azo dye using mixed culture in altering anaerobic/aerobic sequencing batch reactors. Biotechnol Bioprocess Eng 17:875–880CrossRefGoogle Scholar
  16. Jadhav JP, Kalyani DC, Telke AA, Phugare SS, Govindwar SP (2010) Evaluation of the efficacy of a bacterial consortium for the removal of color, reduction of heavy metals, and toxicity from textile dye effluent. Bioresour Technol 10:165–173CrossRefGoogle Scholar
  17. Joe MH, Lim SY, Kim DH, Lee IS (2008) Decolorization of reactive dyes by Clostridium bifermentans SL186 isolated from contaminated soil. World J Microbiol Biotechnol 24:2221–2226CrossRefGoogle Scholar
  18. Kabra AN, Khandarea RV, Waghmode TR, Govindwar SP (2011) Differential fate of metabolism of a sulfonated azo dye Remazol Orange 3R by plants Aster amellus Linn., Glandularia pulchella (sweet) Tronc. And their consortium. J Hazard Mater 190:424–431CrossRefGoogle Scholar
  19. Kalyani DC, Telke AA, Dhanve RS, Jadhav JP (2009) Ecofriendly biodegradation and detoxification of reactive red 2 textile dye by newly isolated Pseudomonas sp. SUK1. J Hazard Mater 163:735–742CrossRefGoogle Scholar
  20. Karim Md E, Dhar K, Md HT (2018) Decolorization of textile reactive dyes by bacterial monoculture and consortium screened from textile dyeing effluent. J Genet Eng Biotechnol 16:375–380CrossRefGoogle Scholar
  21. Kurade MB, Waghmode TR, Jadhav MU, Jeon B, Govindwar SP (2015) Bacterial-yeast consortium as an effective biocatalyst for biodegradation of sulphonated azo dye reactive red 198. RSC Adv 5:23046–23056CrossRefGoogle Scholar
  22. Lowry OH, Rosebrough NJ, Randall AL (1951) Protein measurement with the folinphenol reagent. J Biol Chem 193:265–275Google Scholar
  23. Mishra S, Maiti A (2018) The efficacy of bacterial species to decolourise reactive azo, anthroquinone and triphenylmethane dyes from wastewater: a review. Environ Sci Pollut Res 25:8286–8314CrossRefGoogle Scholar
  24. Moosvi S, Kher X, Madaniwar D (2007) Isolation, characterization and decolorization of textile dyes by a mixed bacterial consortium J W-2. Dyes Pigments 74:723–729CrossRefGoogle Scholar
  25. Nachiyar CV, Rajkumar GS (2003) Degradation of tannery and textile dye, Navitan fast blue S5R by Pseudomonas aeruginosa. World J Microbiol Biotechnol 19:609–614CrossRefGoogle Scholar
  26. Neetha JN, Sandesha K, Girish Kumar K, Chidanandab B, Ujwal P (2019) Optimization of direct Blue-14 dye degradation by Bacillus fermus (Kx898362) an alkaliphilic plant endophyte and assessment of degraded metabolite toxicity. J Hazard Mater 364:742–751CrossRefGoogle Scholar
  27. Oturkar CC, Nemad HN, Mulik PM, Patole MS, Hawaldar KR, Gawai KR (2011) Mechanistic investigation of decolorization and degradation of reactive red 120 by Bacillus lentus BI377. Bioresour Technol 102:758–764CrossRefGoogle Scholar
  28. Oturkar C, Patole MS, Gawai KR, Madmwar D (2013) Enzyme based cleavage strategy of Bacillus lentus BI377 in response to metabolism of azoic recalcitrant. Bioresour Technol 130:360–365CrossRefGoogle Scholar
  29. Padhi S, Tripathy S, Sen R, Mahapatra A, Mohanty S, Maiti N (2013) Characterization of heterotrophic nitrifying and aerobic denitrifying Klebsiella pneumoniae CF-S9 strain for bioremediation of wastewater. Int Biodeterior Biodegrad 78:67–73CrossRefGoogle Scholar
  30. Patil PS, Phugare SS, Jadhav SB (2010) Communal action of microbial cultures for red HE3B degradation. J Hazard Mater 181:263–270CrossRefGoogle Scholar
  31. Pavia DL, Lampman GM, Kriz GS, Vyvyam JR (2015) IntroducGoogle Scholar
  32. Phugare S, Kalyani DC, Patil AV, Jadhav JP (2011) Textile dye decolorization by bacterial consortium and subsequent toxicological analysis of dye and dye metabolites using cytotoxicity, genotoxicity and oxidative stress studies. J Hazard Mater 186:713–723CrossRefGoogle Scholar
  33. Sahasrabudhe MM, Saratale RG, Saratale GD, Pathade GR (2014) Decolorization and detoxification of sulfonated toxic diazo dye C.I. direct red 81 by Enterococcus faecalis YZ 66. J Environ Health Sci Eng 12:151CrossRefGoogle Scholar
  34. Saratale RG, Saratale GD, Chang JS, Govindwar SP (2011) Bacterial decolorization and degradation of azo dyes: a review. J Taiwan Inst Chem Eng 42:138–157CrossRefGoogle Scholar
  35. Saratale RG, Saratale GD, Govindwar SP, Kim DS (2015) Exploiting the efficacy of Lysinibacillus sp. RGS for decolorization and detoxification of industrial dyes, textile effluent and bioreactor studies. J Environ Sci Health A Tox Hazard Subst Environ Eng 50:176–192CrossRefGoogle Scholar
  36. Shah B, Patel A, Madamwar D (2016) Kinetic modelling and community dynamics of microaerophilic treatment of textile dyes containing effluent by consortium VIE6. Environ Process 3:397–411CrossRefGoogle Scholar
  37. Shrinivasan S, Shanmugam G, Surwase S, Jadhav JP, Sadasivam SK (2017) In silico analysis of bacterial systems for textile azo dye decolorization and affirmation with wetlab studies. Clean Soil Air Water 45:1–16Google Scholar
  38. Stieglmeier M, Wirth R, Kminek G, Moissl-Eichinger C (2009) Cultivation of anaerobic and facultatively anaerobic bacteria from spacecraft-associated clean rooms. Appl Environ Microbiol 75:3484–3491CrossRefGoogle Scholar
  39. Surwase SV, Deshpande KK, Phugare SS, Jadhav JP (2013) Biotransformation studies of textile dye Remazol Orange 3R. 3 Biotech 3:267–275CrossRefGoogle Scholar
  40. Tan L, He M, Song L, Fu X, Shi S (2016) Aerobic decolorization, degradation and detoxification of azo dyes by a newly isolated salt-tolerant yeast Scheffersomyces spartinae. TLHS-SF1. Bioresour Technol 203:287–294CrossRefGoogle Scholar
  41. Thakur J, Paul S, Dureja P, Annapurna K, Padaria JC, Gopal M (2014) Degradation of sulphonated azo dye red HE7B by Bacillus sp. and elucidation of degradative pathways. Curr Microbiol 69:183–191CrossRefGoogle Scholar
  42. Tony BD, Goyal D, Khanna S (2009) Decolorization of textile azo dyes by aerobic bacterial consortium. Int Biodeterior Biodegrad 63:462–469CrossRefGoogle Scholar
  43. Walkely A, Black IA (1934) An examination of the Degtjareff method for determining soil organic matter and a proposed modification of the chromic acid titration method. Soil Sci 37:29–38CrossRefGoogle Scholar
  44. Wang H, Su J, Zheng X, Tian Y, Xiong X, Zheng T (2009) Bacterial decolorization and degradation of the reactive dye reactive red 180 by Citrobacter sp. CK3. Int Biodeterior Biodegrad 63:395–399CrossRefGoogle Scholar
  45. Young LY, Phelps CD (2005) Metabolic biomarkers for monitoring in situ anaerobic hydrocarbon degradation. Environ Health Perspect 113:62–67CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Kshama Balapure
    • 1
  • Payal Aghera
    • 1
  • Nikhil Bhatt
    • 1
    Email author
  • Datta Madamwar
    • 2
  1. 1.Department of Microbiology and Biogas Research Extension CentreSadraIndia
  2. 2.Department of Biosciences, UGC Centre of Advanced studySardar Patel UniversityAnandIndia

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