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Properties of Laccase of Bacillus marisflavi Strain BB4 and its Synthetic Dyes Decolorization Analysis

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

In the present study, a bacterium-producing a laccase capable of decolorizing a dye was isolated from Bengaluru soil and identified as Bacillus marisflavi strain BB4 using by 16S rDNA sequencing. The optimum production was seen at 30 °C, pH 7.0 and 48 h when the culture flasks were shaken at 100 rpm with 1% inoculum level. A remarkable overall 5.2-fold increase in laccase production was attained after optimization. After (NH4)2SO4 precipitation and ion exchange technique, the calculated specific activity was 398.2 U/mg. The properties for the purified laccase were 52 kDa, 30 °C, pH 7.0, stimulated by Cu2+, strongly inhibited by Hg2+, no effect with Mg2+, Zn2+, Co2+, Na+, Fe2+, Mn2+, EDTA, inhibited by SDS, NaN3, L-Cys, 2-mercaptoethanol (2-ME), and dithiothreitol, oxidize guaiacol, 2,2′-azino-bis(3-ethylbenzothazoline-6-sulfonate (ABTS), catechol, and 2,6-dimethoxyphenol (2,6-DMP), has 1.43 µM and 227.3 U/ml as Km and Vmax, respectively. The laccase of B. marisflavi was able to decolorize Congo red, bromophenol blue, crystal violet, and indigo carmine without a redox mediator. However, the ABTS was needed as redox mediator to degrade malachite green and reactive orange 16 to a great extent. The laccase of the present study is therefore a good candidate for textile industries as decolorization agent.

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References

  1. Rajeshwari M, Bhuvaneswari V (2016) Production of the extracellular laccase from the newly isolated Bacillus sp. PK4. Afri J Biotechnol 15:1813–1826. https://doi.org/10.5897/AJB2016.15509

    Article  Google Scholar 

  2. Chauhan PS, Goradia B, Saxena A (2017) Bacterial laccase: recent update on production, properties and industrial applications. 3 Biotech https://doi.org/10.1007/s13205-017-0955-7

  3. Allos MM, Hussein AA (2015) Optimum conditions for laccase production by local isolate of Bacillus cereus B5. J Al-Nahrain Uni 18:133–140 https://doi.org/10.13140/RG.2.2.20845.64485

  4. Jeon SJ, Lim SJ (2017) Purification and characterization of the laccase involved in dye decolorization by the white-rot Fungus Marasmius scorodonius. J Microbiol Biotechnol 27:1120–1127. https://doi.org/10.4014/jmb.1701.01004

    Article  CAS  PubMed  Google Scholar 

  5. Mandic M, Djokic L, Nikolaivits E et al (2019) Identification and characterization of new laccase biocatalysts from Pseudomonas species suitable for degradation of synthetic textile dyes. Catalysts 9:6–29. https://doi.org/10.3390/catal9070629

    Article  CAS  Google Scholar 

  6. Kuddus M, Joseph B, Wasudev P (2013) Production of laccase from newly isolated Pseudomonas putida and its application in bioremediation of synthetic dyes and industrial effluents. Biocatal Agric Biotechnol 2:333–338. https://doi.org/10.1016/j.bcab.2013.06.002

    Article  Google Scholar 

  7. Verma A, Dhiman K, Shirkot P (2016) Hyper-production of laccase by Pseudomonas putida LUA15.1 through mutagenesis. J Microbiol Exp 3:21–26. https://doi.org/10.15406/jmen.2016.03.00080

  8. Sondhi S, Sharma P, Saini S, Puri N, Gupta N (2014) Purification and characterization of an extracellular, thermo-alkali-stable, metal tolerant laccase from Bacillus tequilensis SN4. PLoS ONE. https://doi.org/10.1371/journal.pone.0096951

    Article  PubMed  PubMed Central  Google Scholar 

  9. Guo H, Lin C, Wang S et al (2017) Characterization of a novel laccase-producing Bacilllus sp. A4 and its application in Miscanthus degradation. Bioresour 12:4776–4794. https://doi.org/10.15376/biores.12.3.4776-4794

  10. Sondhi S, Saini K (2019) Response surface based optimization of laccase production from Bacillus sp. MSK-01 using fruit juice waste as an effective substrate. Heliyon. https://doi.org/10.1016/j.heliyon.2019.e01718

  11. Kiiskinen LL, Rättö M, Kruus K (2004) Screening for novel laccase-producing microbes. J Appl Microbiol 97:640–646. https://doi.org/10.1111/j.1365-2672.2004.02348.x

    Article  CAS  PubMed  Google Scholar 

  12. Yang Q, Zhang M, Zhang M et al (2018) Characterization of a novel, cold-adapted, and thermostable laccase-like enzyme with high tolerance for organic solvents and salt and potent dye decolorization ability, derived from a marine metagenomic library. Front Microbiol 9:1–9. https://doi.org/10.3389/fmicb.2018.02998

    Article  Google Scholar 

  13. Bhuvaneshwari V, Preethikaharshini J, Amsaveni R, Kalaiselvi M (2015) Isolation, optimization and production of laccase from Halobacillus halophilus. Int J Biosci Nanosci 2:41–47

    Google Scholar 

  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    Article  CAS  Google Scholar 

  15. Blum H, Beier H, Gross HJ (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8:93–99. https://doi.org/10.1002/elps.1150080203

    Article  CAS  Google Scholar 

  16. Niyonzima FN, More SS (2015) Purification and characterization of detergent compatible alkaline protease from Aspergillus terreus gr. 3 Biotech 5:61–70. https://doi.org/10.1007/s13205-014-0200-6

  17. Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666

  18. Kumar M, Mishra A, Singh SS, Srivastava S, Thakur IS (2018) Expression and characterization of novel laccase gene from Pandoraea sp. ISTKB and its application. Int J Biol Macromol 115:308–316. https://doi.org/10.1016/j.ijbiomac.2018.04.079

    Article  CAS  PubMed  Google Scholar 

  19. Mehandia S, Sharma SC, Arya SK (2020) Isolation and characterization of an alkali and thermostable laccase from a novel Alcaligenes faecalis and its application in decolorization of synthetic dyes. Biotechnol Rep. https://doi.org/10.1016/j.btre.2019.e00413

    Article  Google Scholar 

  20. Niyonzima FN, Veena SM, More SS (2020) Industrial production and optimization of microbial enzymes. In: Arora NA, Mishra J, Mishra V (Eds.), Microbial enzymes: Roles and applications in industries. Springer, Singapore, pp 115–135. https://doi.org/10.1007/978-981-15-1710-5_5

  21. Niyonzima FN, More SS (2015) Coproduction of detergent compatible bacterial enzymes and stain removal evaluation. J Basic Microbiol 55:1–10. https://doi.org/10.1002/jobm.201500112

    Article  CAS  Google Scholar 

  22. Niladevi KN, Sukumaran RN, Prema P (2007) Utilization of rice straw for laccase production by Streptomyces psammoticus in solid-state fermentation. J Ind Microbiol Biotechnol 34:665–674. https://doi.org/10.1007/s10295-007-0239-z

    Article  CAS  PubMed  Google Scholar 

  23. Fonseca MI, Shimizu E, Zapata PD, Villalba LL (2010) Copper inducing effect on laccase production of white rot fungi native from Misiones (Argentina). Enzyme Microb Tech 46:534–539. https://doi.org/10.1016/j.enzmictec.2009.12.017

    Article  CAS  Google Scholar 

  24. Mathews SL, Smithson CE, Grunden AM (2016) Purification and characterization of a recombinant laccase-like multi-copper oxidase from Paenibacillus glucanolyticus SLM1. J Appl Microbiol 121:1335–1345. https://doi.org/10.1111/jam.13241

    Article  CAS  PubMed  Google Scholar 

  25. Niyonzima FN (2019) Detergent compatible bacterial cellulases. J Basic Microbiol 59:134–147. https://doi.org/10.1002/jobm.201800436

    Article  CAS  PubMed  Google Scholar 

  26. Li Y, Zuo W, Li Y, Wang X (2012) Cloning of multicopper oxidase gene from Ochrobactrum sp. 531 and characterization of its alkaline laccase activity towards phenolic substrates. Adv Biol Chem 2:248–255. https://doi.org/10.4236/abc.2012.23031

    Article  CAS  Google Scholar 

  27. Zheng Z, Li H, Li L, Shao W (2012) Biobleaching of wheat straw pulp with recombinant laccase from the hyperthermophilic Thermus thermophiles. Biotechnol Lett 34:541–547. https://doi.org/10.1007/s10529-011-0796-0

    Article  CAS  PubMed  Google Scholar 

  28. Galletti P, Pori M, Funiciello F, Soldati R, Ballardini A, Giacomini D (2014) Laccase-mediator system for alcohol oxidation to carbonyls or carboxylic acids: toward a sustainable synthesis of profens. Chem Sus Chem 7:2684–2689. https://doi.org/10.1002/cssc.201402136

    Article  CAS  Google Scholar 

  29. Lu L, Zhao M, Wang NY et al (2012) Characterization and dye decolorization ability of an alkaline resistant and organic solvents tolerant laccase from Bacillus licheniformis LS04. Bioresour Technol 115:35–40. https://doi.org/10.1016/j.biortech.2011.07.111

    Article  CAS  Google Scholar 

  30. Zhang C, Diao H, Lu F, Bie X, Wang Y, Lu Z (2012) Degradation of triphenylmethane dyes using a temperature and pH stable spore laccase from a novel strain of Bacillus vallismortis. Bioresour Technol 126:80–86. https://doi.org/10.1016/j.biortech.2012.09.055

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

All the authors would like to thank the Management of Dayananda Sagar University for all the necessary facilities. All authors declare that no financial/commercial conflicts of interest

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Authors and Affiliations

  1. School of Basic and Applied Sciences, Dayananda Sagar University, Bangalore, 560111, India

    Anu Sharma, M. Muthupriya, Radharaman Raj, Zainah Shameen & Sunil S. More

  2. Department of Biotechnology, Sapthagiri College of Engineering, Bangalore, 560090, India

    Veena SM

  3. Department of Biotechnology, INES-Ruhengeri, Musanze-155, Ruhengeri, Rwanda

    Francois N. Niyonzima

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  1. Anu Sharma
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  3. Radharaman Raj
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  6. Francois N. Niyonzima
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  7. Sunil S. More
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Correspondence to Sunil S. More.

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Significance Statement

Although synthetic dyes are mostly utilized in industries, they cause pollution and may act as carcinogenic agents. Use of biodegradable macromolecules like laccases is needed nowadays. The search for laccase with desirable properties is a continuous task since the existing ones are produced at high cost and not principally commercialized. Here, laccase of Bacillus marisflavi was characterized as decolorization agent.

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Sharma, A., Muthupriya, M., Raj, R. et al. Properties of Laccase of Bacillus marisflavi Strain BB4 and its Synthetic Dyes Decolorization Analysis. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 91, 477–485 (2021). https://doi.org/10.1007/s40011-021-01235-0

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