Skip to main content
SpringerLink
Log in

Isolation and characterization of alkalotolerant bacteria and optimization of process parameters for decolorization and detoxification of pulp and paper mill effluent by Taguchi approach

  • Original Paper
  • Published:
Biodegradation Aims and scope Submit manuscript

Abstract

Four different bacterial strains were isolated from pulp and paper mill sludge in which one alkalotolerant isolate (LP1) having higher capability to remove color and lignin, was identified as Bacillus sp. by 16S RNA sequencing. Optimization of process parameters for decolorization was initially performed to select growth factors which were further substantiated by Taguchi approach in which seven factors, % carbon, % black liquor, duration, pH, temperature, stirring and inoculum size, at two levels, applying L-8 orthogonal array were taken. Maximum color was removed at pH 8, temperature 35°C, stirring 200 rpm, sucrose (2.5%), 48 h, 5% (w/v) inoculum size and 10% black liquor. After optimization 2-fold increase in color and lignin removal from 25–69% and 28–53%, respectively, indicated significance of Taguchi approach in decolorization and delignification of lignin in pulp and paper mill effluent. Enzymes involved in the process of decolorization of effluent were found to be xylanase (54 U/ml) and manganese peroxidase (28 U/ml). Treated effluent was also evaluated for toxicity by Comet assay using Saccharomyces cerevisiae MTCC 36 as model organism, which indicated 58% reduction after treatment by bacterium.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Adikane HV, Dange MN, Selvakumari K (2006) Optimization of anaerobically digested distillery molasses spent wash decolorization using soil as inoculum in the absence of additional carbon and nitrogen source. Biores Technol 97:2131–2135

    Article  CAS  Google Scholar 

  • Ali M, Sreekrishnan TR (2001) Aquatic toxicity from pulp and paper mill effluent: a review. Adv Environ Res 5:175–196

    Article  CAS  Google Scholar 

  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DG (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    CAS  PubMed  Google Scholar 

  • APHA (2005) Standard methods for the examination of water and wastewater, 21st edn. American Public Health Association, Washington

    Google Scholar 

  • Bajpai P, Mehna A, Bajpai PK (1993) Decolorization of Kraft bleach plant effluent with the white rot fungus Trametes versicolor. Proc Biochem 28:377–384

    Article  CAS  Google Scholar 

  • Bakhtiari MR, Faezi MG, Fallahpour M, Noohi A, Moazami N, Amidi Z (2006) Medium optimization by orthogonal array designs for urease production by Aspergillus niger PTCC5011. Proc Biochem 41:547–551

    Article  CAS  Google Scholar 

  • Bourbonnais R, Paice MG (1992) Demethylation and delignification of Kraft pulp by Trametes versicolor laccase in the presence of 2, 2-azinobis-3-ethylbenzthiazoline-6-sulphonate. Appl Microbiol Biotechnol 36:823–827

    Article  CAS  Google Scholar 

  • Crooks R, Sikes J (1990) Environmental effects of bleached Kraft mill effluents. Appita 43:67–76

    CAS  Google Scholar 

  • Dasu VV, Panda T, Chidambaram M (2003) Determination of significant parameters for improved griseofulvin production in a batch bioreactor by Taguchi’s method. Proc Biochem 38:877–880

    Article  Google Scholar 

  • El-Bestawy E, El-Sokkary I, Hussein H, Keela AFA (2008) Pollution control in pulp and paper industrial effluents using integrated chemical–biological treatment sequences. J Ind Microbiol Biotechnol 35:1517–1529

    Article  CAS  PubMed  Google Scholar 

  • Giovannoni SJ, Britschgi TB, Moyer CL, Field KG (1990) Genetic diversity in Sargasso Sea bacterioplankton. Nature 345:60–63

    Article  CAS  PubMed  Google Scholar 

  • Jadhav SU, Jadhav MU, Kagalkar AN, Govindwar SP (2008) Decolorization of Brilliant Blue G dye mediated by degradation of the microbial consortium of Galactomyces geotrichum and Bacillus sp. J Chin Inst Chem Eng 39(6):563–570

    Article  CAS  Google Scholar 

  • Kackar R (1985) Off-line quality control, parameter design and Taguchi method. J Qual Technol 17:176–188

    Google Scholar 

  • Kinae N, Hashu T, Makita T, Tomita I, Kimura I, Kanamori H (1981) Studies on the toxicity of pulp and paper Mill effluents: Mutagenicity of the sediment samples derived from Kraft paper mills. Water Res 15:17–24

    Article  Google Scholar 

  • Kirk TK, Connors WJ, Zeikus JG (1977) Advances in understanding the microbiological degradation of lignin. Rev Adv Phytopathol 11:369–394

    CAS  Google Scholar 

  • Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment brief. Bioinformatics 5:150–163

    Article  CAS  PubMed  Google Scholar 

  • Lindstrom-Seppa P, Hunskonen S, Kotelevtsev S, Mikkelson P, Rannen T, Stepanova L, Hanninena O (1998) Toxicity and mutagenity of waste waters from Baikalsk pulp and paper mill: evaluation of pollutant contamination in lake Baikal. Mar Environ Res 46:273–277

    Article  CAS  Google Scholar 

  • Miloshev G, Mihaylov I, Anachkova B (2002) Application of the single cell gel electrophoresis on yeast cells. Mutat Res 513:69–74

    CAS  PubMed  Google Scholar 

  • Miyamae Y, Yamamoto M, Sasaki YF, Kobayashi H, Igarashi-Soga M, Shimoi K, Hayashi M (1998) Evaluation of a tissue homogenization technique that isolates nuclei for the in vivo single cell gel electrophoresis comet assay: a collaborative study by five laboratories. Mutat Res 418:131–140

    CAS  PubMed  Google Scholar 

  • Moriya RY, Gonçalves AR, Duarte MCT (2005) Sugarcane bagasse pulps: biobleaching with commercial catarzyme HS and with Bacillus pumilus xylanase. Appl Biochem Biotechnol 121:171–181

    Article  PubMed  Google Scholar 

  • Nagarathnamma R, Bajpai P (1999) Decolorization and detoxification of extraction-stage effluent from chlorine bleaching of Kraft pulp by Rhizopus oryzae. Appl Environ Microbiol 65(3):1078–1082

    CAS  PubMed  Google Scholar 

  • Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 153:375–380

    CAS  Google Scholar 

  • Niku-Paavola ML, Karhunen E, Salola P, Raunio V (1988) Ligninolytic enzymes of the white-rot fungus Phlebia radiata. Biochem J 254:877–884

    CAS  PubMed  Google Scholar 

  • Oliveira PL, Duarte MCT, Ponezi AN, Durrant LR (2009) Use of Bacillus pumilus CBMAI 0008 and Paenibacillus sp. CBMAI 868 for colour removal from paper mill effluent. Braz J Microbiol 40:354–357

    Google Scholar 

  • Pearl IA, Benson HK (1940) The determination of lignin in sulphide pulping liquor. Pap Trade J 111:35–36

    CAS  Google Scholar 

  • Phadke MS, Dehnad K (1988) Optimization of product and process design for quality and cost. Qual Reliab Eng Int 4:159–169

    Article  Google Scholar 

  • Pokhrel D, Viraraghavan T (2004) Treatment of pulp and paper mill wastewater: a review. Sci Total Environ 333:37–58

    Article  CAS  PubMed  Google Scholar 

  • Prakasham RS, Rao CS, Rao S, Rajesham S, Sharma PN (2005) Optimization of alkaline protease production by Bacillus sp. using Taguchi methodology. Appl Biochem Biotechnol 120:133–144

    Article  CAS  PubMed  Google Scholar 

  • Ratanakhanokchai K, Kyu KL, Tanticharoen M (1999) Purification and properties of a xylan-binding endoxylanase from alkaliphilic Bacillus sp. strain K-1. Appl Environ Microbiol 65:694–697

    CAS  PubMed  Google Scholar 

  • Roy RK (2001) Design of experiments using the Taguchi approach: 16 steps to product and process improvement. Wiley, New York

    Google Scholar 

  • Roy RK (2007) Qualitek-4, software for automatic design and analysis of Taguchi experiments. Nutek Inc., Bloomfield Hills

    Google Scholar 

  • Schnell A, Steel P, Melcer H, Hodson PV, Carey JH (2000) Enhanced biological treatment of bleached kraft mill effluents: II. Reduction of mixed function oxygenase (MFO) induction in fish. Water Res 34:501–509

    Article  CAS  Google Scholar 

  • Scortichini M, Marchesi U, Rossi MP, Di Prospero P (2002) Bacteria associated with Hazelnut (Corylus avellana L.) decline are of two groups: Pseudomonas avellanae and strains resembling P. syringae pv. Syringae. Appl Environ Microbiol 68:476–484

    Article  CAS  PubMed  Google Scholar 

  • Sirianuntapiboon S, Zohsalam P, Ohmomo S (2004) Decolorization of molasses wastewater by Citeromyces sp. WR-43-6. Proc Biochem 39:917–924

    Article  CAS  Google Scholar 

  • Stowe RA, Mayer RP (1999) Efficient screening of process variables. Ind Eng Chem 56:36–40

    Google Scholar 

  • Thakur IS (2004) Screening and identification of microbial strains for removal of colour and adsorbable organic halogens in pulp and paper mill effluent. Proc Biochem 39:1693–1699

    Article  CAS  Google Scholar 

  • Thakur IS, Verma P, Upadhayaya KC (2001) Involvement of plasmid in degradation of pentachlorophenol by Pseudomonas sp. from a chemostat. Biochem Biophys Res Commun 286:109–113

    Article  CAS  PubMed  Google Scholar 

  • Tien M, Kirk TK (1988) Lignin peroxidase of Phanerochaete chrysosporium. Methods Enzymol 161:238–249

    Article  CAS  Google Scholar 

  • Vass KK, Mukopadhyay MK, Mistra K, Joshi HC (1996) Respiratory stresses in fishes exposed to paper and pulp wastewater. Environ Ecol 14:895–897

    Google Scholar 

  • Vicuna R (1988) Bacterial degradation of lignin. Enzyme Microbiol Technol 10:646–655

    Article  CAS  Google Scholar 

  • Vicuna R, Gonzalez B, Seelenfreund D, Ruttimann C, Salas L (1993) Ability of natural bacterial isolates to metabolize high and low molecular weight lignin-derived molecules. J Biotechnol 30:9–13

    Article  CAS  Google Scholar 

  • Wariishi H, Valli K, Gold MH (1992) Manganese (II) oxidation by manganese peroxidase from the basidiomycete Phanerochaete chrysosporium: kinetic mechanism and role of chelators. J Biol Chem 267:23688–23695

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This paper was supported by the research grants of Department of Biotechnology, Government of India, and New Delhi, India. The author (MM) thanks Council for Scientific and Industrial Research, Government of India, New Delhi, India, for providing Research Fellowship. We also thank Century pulp and paper mill, Uttarakhand, India for providing effluent and sludge during the course of investigation and R. K. Roy for providing Qualitek-4 software.

Author information

Authors and Affiliations

  1. School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, 110067, India

    Monika Mishra & Indu Shekhar Thakur

Authors
  1. Monika Mishra
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Indu Shekhar Thakur
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Monika Mishra.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mishra, M., Thakur, I.S. Isolation and characterization of alkalotolerant bacteria and optimization of process parameters for decolorization and detoxification of pulp and paper mill effluent by Taguchi approach. Biodegradation 21, 967–978 (2010). https://doi.org/10.1007/s10532-010-9356-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10532-010-9356-x

Keywords

Search

Navigation