Production of laccase by repeated batch semi-solid fermentation using wheat straw as substrate and support for fungal growth
Repeated batch semi-solid fermentation (sSF) process using wheat straw substrate and fungal growth of Ganoderma lucidum on solid substrate was studied for production of laccase. pH showed significant effect on laccase production. Highest laccase activity with pH controlled to 5.0 in batch sSF was 15257.2 ± 353.4 U L− 1 on 9th day. In repeated batch process at pH 5.0, insoluble biomass substrate and fungal growth were reused after liquid part of medium was replaced with glucose, ammonium phosphate (best nitrogen source) and combined glucose and ammonium phosphate solution separately. Refilled to 80% w v− 1 of initial soluble sugar of first batch resulted in highest laccase production with peak activity after 4 days from replacement. Production of enzyme increased from 15257.2 U L− 1 in first batch to cumulative 90164.4 U L− 1 in 29 days after six repeated batches, productivity increased from 1680.2 to 3110.3 U L− 1 day− 1 (∼ 1.9 times) due to reductions in inhibitory effects and time required for fungal growth. Utilization of wheat straw in repeated batch sSF was supported by composition analysis and morphological changes (scanning electron microscopy) of substrate. Economic production of laccase using agricultural residues in repeated batch sSF could be possible.
KeywordsWhite rot fungi Semi-solid fermentation Repeated batch process Laccase production
Mr Antriksh Gupta gratefully acknowledges Ministry of Human Resource Development (MHRD), Govt. of India, for providing the fellowship during the study. All authors are highly thankful to National Institute of Technology (NIT), Jalandhar, for providing grants and administrative supports for the study.
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Conflict of interest
Authors declare that they have no conflict of interest.
- 6.Gonzalez JC, Medina SC, Rodriguez A, Osma JF, Alméciga-Díaz CJ, Sánchez OF (2013) Production of Trametes pubescens laccase under submerged and semi-solid culture conditions on agro-industrial wastes. PLoS One 8(9):e73721–e73735. https://doi.org/10.1371/journal.pone.0073721 CrossRefPubMedPubMedCentralGoogle Scholar
- 7.Parenti A, Muguerza E, Iroz AR, Omarini A, Conde E, Alfaro M, Castanera R, Santoyo F, Ramírez L, Pisabarro AG (2013) Induction of laccase activity in the white rot fungus Pleurotus ostreatus using water polluted with wheat straw extracts. Bioresour Technol 133:142–149. https://doi.org/10.1016/j.biortech.2013.01.072 CrossRefPubMedGoogle Scholar
- 9.Hakala TK (2007) Characterization of the lignin-modifying enzymes of the selective white-rot fungus Physisporinus rivulosus, vol 1. Helsinki University Printing House, Helsinki, pp 1–60Google Scholar
- 23.Osma JF, Moilanen U, Toca-Herrera JL, Rodríguez-Couto S (2011) Morphology and laccase production of white-rot fungi grown on wheat bran flakes under semi-solid-state fermentation conditions. FEMS Microbiol Lett 318(1):27–34. https://doi.org/10.1111/j.1574-6968.2011.02234.x CrossRefPubMedGoogle Scholar
- 29.Pointing SB (1999) Qualitative methods for the determination of lignocellulolytic enzyme production by tropical fungi. Fungal Divers 2(3):17–33Google Scholar
- 34.Sluiter A, Hames B, Ruiz R, Scarlata C, Sluiter J, Templeton D, Crocker D (2010) Determination of structural carbohydrates and lignin in biomass: laboratory analytical procedure (LAP). National Renewable Energy Laboratory, GoldenGoogle Scholar
- 39.Sitarz AK, Mikkelsen JD, Hojrup P, Meyer AS (2013) Identification of a laccase from Ganoderma lucidum CBS 229. 93 having potential for enhancing cellulase catalyzed lignocellulose degradation. Enzyme Microbial Technol 53(6–7):378–385. https://doi.org/10.1016/j.enzmictec.2013.08.003 CrossRefGoogle Scholar
- 42.Philippoussis A, Diamantopoulou P, Papadopoulou K, Lakhtar H, Roussos S, Parissopoulos G, Papanikolaou S (2010) Biomass, laccase and endoglucanase production by Lentinula edodes during solid state fermentation of reed grass, bean stalks and wheat straw residues. World J Microbiol Biotechnol 27(2):285–297. https://doi.org/10.1007/s11274-010-0458-8 CrossRefGoogle Scholar
- 43.Shi J, Chinn MS, Sharma-Shivappa RR (2014) Interactions between fungal growth, substrate utilization, and enzyme production during solid substrate cultivation of Phanerochaete chrysosporium on cotton stalks. Bioprocess Biosyst Eng 37(12):2463–2473. https://doi.org/10.1007/s00449-014-1224-3 CrossRefPubMedGoogle Scholar
- 56.Stajić M, Persky L, Friesem D, Hadar Y, Wasser SP, Nevo E, Vukojević J (2006) Effect of different carbon and nitrogen sources on laccase and peroxidases production by selected Pleurotus species. Enzyme Microbial Technol 38(1–2):65–73. https://doi.org/10.1016/j.enzmictec.2005.03.026 CrossRefGoogle Scholar
- 57.Kenkebashvili N, Elisashvili V, Wasser SP (2012) Effect of carbon, nitrogen sources, and copper concentration on the ligninolytic enzyme production of Coriolopsis gallica. J Waste Convers Bioprod Biotechnol 1(2):22–27Google Scholar