Acta Scientific Microbiology (ISSN: 2581-3226)

Research Article Volume 6 Issue 6

Screening and Identification of Novel Laccase Producing Entomopathogenic Fungus Beauveria pseudobassiana PHF4 and Optimization of Production Conditions of Enzyme

Divya Sharma1, Manpreet Kaur1, Sunita Borkar2 and Reena Gupta1*

1Department of Biotechnology, Himachal Pradesh University, Summerhill, Shimla, India
2Department of Microbiology, PES College of Arts and Science, Farmagudi, Ponda, Goa, India

*Corresponding Author: Reena Gupta, Professor, Department of Biotechnology, Himachal Pradesh University, Summerhill, Shimla, India. E-mail: reenagupta_2001@yahoo.com.

Received: May 08, 2023; Published: May 19, 2023

Abstract

In the present study, twenty-nine positive fungal isolates for production of laccase were obtained from decaying wood and soil samples from Shimla district of Himachal Pradesh, India. The positive isolates were confirmed by brown halo formation owing to oxidation of guaiacol by laccase enzyme. Out of the 29 isolates, the isolate PHF4, which produced the maximum laccase activity, was recognized as Beauveria pseudobassiana. Laccase displayed good activity in production medium comprising peptone (1.0%, w/v), malt extract (0.5%, w/v), glucose (2.0%, w/v) and CuSO4 (0.005%, w/v). Optimization of inoculum size, incubation time, carbon source, carbon source concentration, nitrogen source, nitrogen source concentration, copper sulphate concentration, temperature and pH for laccase production was performed. The optimal inoculum size and incubation time for the laccase production were found to be 6 discs (1 cm in diameter) and 8 days respectively. Glucose (2.0%, w/v) and yeast extract (2.0%, w/v) were the most appropriate carbon and nitrogen source respectively for the production of enzyme. The presence of 0.003% (w/v) copper sulphate enhanced the laccase production. A temperature of 25°C and pH 6.0 were found to be optimum for laccase production. Under the optimum conditions, the maximum activity of enzyme was 21.47 U/ml which was 2.9 times higher than the unoptimized conditions. This is the first record of Beauveria pseudobassiana being isolated and optimized for the production of laccase.

 Keywords: Laccase; Optimization; Beauveria pseudobassiana; Guaiacol; Yeast Extract

References

  1. Devasia S and Nair AJ. “Screening of potent laccase producing organisms based on the oxidation pattern of different phenolic substrates”. International Journal of Current Microbiology and Applied Sciences 5 (2016): 127-137.
  2. Mehra R., et al. “A structural chemical explanation of fungal laccase activity”. Scientific Reports 8 (2018): 17285.
  3. Surwase SV., et al. “Interaction of small molecules with fungal laccase: a surface plasmon resonance-based study”. Enzyme and Microbial Technology 82 (2016): 110-114.
  4. Agrawal K., et al. “Fungal laccase discovered but yet undiscovered”. Bioresources and Bioprocessing 5 (2018): 1-12.
  5. Pooalai R., et al. “Genomic analysis uncovers laccase-coding genes and biosynthetic gene clusters encoding antimicrobial compounds in laccase-producing Acinetobacter baumannii”. Scientific Reports 12 (2022): 11932.
  6. Janusz G., et al. “Laccase properties, physiological functions, and evolution”. International Journal of Molecular Sciences 21 (2020): 966.
  7. Christopher LP., et al. “Lignin Biodegradation with Laccase-Mediator Systems”. Frontiers in Energy Research 2 (2014): 13.
  8. Mate DM and Alcalde M. “Laccase engineering: from rational design to directed evolution”. Biotechnology Advances 33 (2015): 25-40.
  9. Pourkhanali K., et al. “Isolation, identification and optimization of enhanced production of laccase from Galactomyces geotrichumunder solid-state fermentation”. Preparative Biochemistry and Biotechnology 51 (2021): 659-668.
  10. Dong SJ., et al. “Efficient lignin degradation of corn stalk by Trametes with high laccase activity and enzymatic stability in salt and ionic liquid”. Bioresources 14 (2019): 5339-5354.
  11. Arregui L., et al. “Laccases: structure, function and potential application in water bioremediation”. Microbial Cell Factories 18 (2019):
  12. Xenakis A., et al. “Enzymatic reactions in structured surfactant-free microemulsions”. Current Opinion in Colloid and Interface Science 22 (2016): 41-45.
  13. Akpinar M and Urek RO. “Induction of fungal laccase production under solid state bioprocessing of New agroindustrial waste and its application on dye decolorization”. 3 Biotech 7 (2017): 98.
  14. Góralczyk-Bińkowska A., et al. “Characteristics and use of multicopper oxidases enzymes”. Postępy Mikrobiologii 58 (2019): 7-18.
  15. Rao A., et al. “Isolation, screening and optimization of laccase-producing endophytic fungi from Euphorbia milii”. Arabian Journal for Science and Engineering 44 (2019): 51-64.
  16. Kumar A., et al. “Fungal phytoremediation of heavy metal-contaminated resources: current scenario and future prospects”. In: Yadav, AN, Singh, S, Mishra, S, Gupta, A. (eds): Recent advancement in white biotechnology through fungi. Switzerland: Springer Nature, (2019): 437-461.
  17. Fillat Ú., et al. “Screening of eucalyptus wood endophytes for laccase activity”. Process Biochemistry51 (2016): 589-598.
  18. Sidhu AK., et al. “Isolation, screening and optimization for laccase production by Scytalidium lignicola pesante under submerged fermentation”. International Journal of Current Microbiology and Applied Sciences 6 (2017): 2477-2491.
  19. Vantamuri AB., et al. “Isolation and characterization of laccase producing fungi from different environmental samples”. International Journal of Recent Scientific Research 6 (2015): 6853-6857.
  20. Sehgal R and Gupta R. “Characterization of medium chain length polyhydroxyalkanoates from Priestia megaterium POD1”. Research Journal of Chemistry and Environment 26 (2022): 46-54.
  21. Kalra K., et al. “Isolation of laccase producing Trichoderma and effect of pH and temperature on its activity”. International Journal of Environmental Science and Technology 5 (2013): 2229-2235.
  22. Minussi RC., et al. “Potential applications of laccase in the food industry”. Trends in Food Science and Technology 13 (2002): 205-216.
  23. Uthandi S., et al. “LccA, an archaeal laccase secreted as a highly stable glycoprotein into the extracellular medium by Haloferax volcanii”. Applied and Environmental Microbiology76 (2010): 733-743.
  24. Senthivelan T., et al. “Screening and production of a potential extracellular fungal laccase from Penicillium chrysogenum, media optimization by response surface methodology (RSM) and central composite rotatable design (CCRD)”. BiotechnologyReports 23 (2019): 30-44.
  25. Sharma RK., et al. “Synthesis, characterization and properties of Mn-doped ZnO nanocrystals”. Advances in Natural Sciences: Nanoscience and Nanotechnology 3 (2012): 35-50.
  26. Dı́az S and Cabido M. “Plant functional diversity matters to ecosystem processes”. Trends in Ecology and Evolution 16 (2001): 646-655.
  27. Luke G., et al. “Analysis of the aphthovirus 2A/2B polyprotein cleavage mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal skip”. Journalof General Virology 82 (2001): 1013-1025.
  28. Liu M., et al. “Thickness-dependent mechanical properties of polydimethylsiloxane membranes”. Journal of Micromechanics and Microengineering 19 (2009): 35-38.
  29. Kuhar F and Papinutti L. “Optimization of laccase production by two strains of Ganoderma lucidum using phenolic and metallic inducers”. Revista Argentina de Microbiologia 46 (2014): 144-149.
  30. Chmelová, et al. “The production of laccases by white-rot fungi under solid-state fermentation conditions”. World Journal of Microbiology and Biotechnology 38 (2022): 1-20.
  31. Rehan A., et al. “Production of laccase enzyme for their potential application to decolorize fungal pigments on aging paper and parchment”. Annals of Agricultural Sciences 61 (2016): 145-154.
  32. Robak K and Balcerek M. “Review of second-generation bioethanol production from residual biomass”. Food Technology and Biotechnology 56 (2018): 174.
  33. Abdul RA., et al. “Production of laccase enzyme for their potential application to decolorize fungal pigments on aging paper and parchment”. Annals of Agricultural Sciences 61 (2016): 145-154.
  34. Brijwani K., et al. “Fungal laccases: production, function, and applications in food processing”. Enzyme Research 3 (2010): 112-113
  35. Pillet L., et al. “Copper-induced production of laccases for lignin depolymerization and micropollutant degradation by laccase-mediator systems”. International Journal of Chemistry 75 (2021): 1058-1065.
  36. Strong PJ. “Improved laccase production by Trametes pubescens MB89 in distillery wastewaters”. Enzyme Research 2 (2011): 6-7.
  37. Reyes C., et al. “Enzyme activities of five white-rot fungi in the presence of nanocellulose”. Journal of Fungus 7 (2021): 222.
  38. Shukur YN. “Determination of optimal conditions for the production of laccase enzyme by local isolate of Bacillus sp”. Iraqi Journal of Science 56 (2015): 132-139.
  39. Thakkar AT and Bhatt SA. “Isolation, identification and optimization of laccase from Alternaria alternate”. Journal of Applied Biology and Biotechnology 8 (2020): 64-69.
  40. Zhang J., et al. “Cancer cell metabolism: the essential role of the nonessential amino acid, glutamine”. EMBO Journal 36 (2017): 1302-1315.
  41. Piscitelli A., et al. “Induction and transcriptional regulation of laccases in fungi”. Current Genomics 12 (2011): 104-112.
  42. Virk AP., et al. “Use of laccase in pulp and paper industry”. Biotechnology progress 28 (2012): 21-32.
  43. Umar A and Ahmed S. “Optimization, purification and characterization of laccase from Ganoderma leucocontextum along with its phylogenetic relationship”. Scientific Reports12 (2022): 1-14.
  44. Jasińska A., et al. “Novel laccase-like multicopper oxidases from the Myrothecium roridum fungus-production enhancement, identification and application in the dye removal process”. Acta Biochimica Polonica 65 (2018): 287-295.
  45. Shrestha P., et al. “Isolation and physicochemical characterization of laccase fromGanoderma lucidum-CDBT1 isolated from its native habitat in Nepal”. BioMed Research International (2016): 3238909.
  46. Ademakinwa AN. “A heat-resistant intracellular laccase immobilized via cross-linked enzyme aggregate preparation: characterization application in bisphenol a removal and phytotoxicity evaluation”. Journal of Hazardous Materials 419 (2021): 126-480.
  47. Belhamiche , et al. “Effect of some agro-industrial residues on mycelial growth and production of lignocellulolytic enzymes by a fungus native to Algerian forest”. Environmental Pollution 1 (2021): 10-11.
  48. Sondhi S and Saini K. “Response surface-based optimization of laccase production from Bacillus sp. MSK-01 using fruit juice waste as an effective substrate”. Heliyon 5 (2019): 17-18.

Citation

Citation: Divya Sharma., et al. “Screening and Identification of Novel Laccase Producing Entomopathogenic Fungus Beauveria pseudobassiana PHF4 and Optimization of Production Conditions of Enzyme". Acta Scientific Microbiology 6.6 (2023): 51-60.

Copyright

Copyright: © 2023 Reena Gupta., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.




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