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Research article
Environmental Microbiology and Engineering
Media Optimization for Laccase Production by Trichoderma harzianum ZF-2 Using Response Surface Methodology
College of Forestry, Shandong Agricultural University, Shandong 271018, China
Received: February 25, 2013; Accepted: September 12, 2013
J. Microbiol. Biotechnol. 2013; 23(12): 1757-1764
Published December 28, 2013 https://doi.org/10.4014/jmb.1302.02057
Copyright © The Korean Society for Microbiology and Biotechnology.
Abstract
Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from
Shandong, China, and showed dye decolorization activities. The objective of this study was to
optimize its culture conditions using a statistical analysis of its laccase production. The
interactions between different fermentation parameters for laccase production were
characterized using a Plackett-Burman design and the response surface methodology. The
different media components were initially optimized using the conventional one-factor-at-atime
method and an orthogonal test design, and a Plackett-Burman experiment was then
performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal,
and CuSO4 were all found to have a significant influence on laccase production, and the
optimal concentrations of these three factors were then sequentially investigated using the
response surface methodology with a central composite design. The resulting optimal medium
components for laccase production were determined as follows: wheat straw powder 7.63 g/l,
soybean meal 23.07 g/l, (NH4)2SO4 1 g/l, CuSO4 0.51 g/l, Tween-20 1 g/l, MgSO4 1 g/l, and
KH2PO4 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased
59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized
medium. This is the first report on the statistical optimization of laccase production by
Trichoderma harzianum ZF-2.
Keywords
Trichoderma harzianum ZF-2, media optimization, laccase production, response surface methodology
References
- Abadulla E, Tzanov T, Costa S, Robra KH, Cavaco PA, Gubitz G. 2000. Decolorization and detoxication of textile dyes with a laccase from Trametes hirsuta. Appl. Environ. Microbiol. 66: 3357-3362.
- Assavanig A, Amornkitticharoen B, Ekpaisal N, Meevootisom V, Flegel TM. 1992. Isolation, characterization and function of laccase from Trichoderma. Appl. Microbiol. Biotechnol. 38:198-202.
- Bollag JM, Leonowicz A. 1984. Comparative studies of extracellular fungal laccases. Appl. Environ. Microbiol. 48:849-854.
- Box GEP, Wilson KB. 1951. On the experimental attainment of optimum conditions. J. Roy. Stat. Soc. B 13: 145.
- Chen T, Barton SC, Binyamin G, Gao Z, Zhang Y, Kim HH, et al. 2001. A miniature biofuel cell. J. Am. Chem. Soc. 123:8630-8631.
- Collins PJ, Dobson ADW. 1997. Regulation of laccase gene transcription in Trametes versicolor. Appl. Environ. Microbiol. 63: 3444-3450.
- Dittmer JK, Patel NJ, Dhawale SW, Dhawale SS. 1997. Production of multiple laccase isoforms by Phanerochaete chrysosporium grown under nutrient sufficiency. FEMS Microbiol. Lett. 149: 65-70.
- Eggert C, Temp U, Eriksson KEL. 1996. The ligninolytic system of the white-rot fungus Pycnoporus cinnabarinus: purification and characterization of the laccase. Appl. Environ. Microbiol. 62: 1151-1158.
- Gao HJ, Wang YW, Zhang WT, Wang WL, Mu ZM. 2011. Isolation, identification and application in lignin degradation of an ascomycete GHJ-4. Afr. J. Biotechnol. 10: 4166-4174.
- Gianfreda L, Xu F, Bollag JM. 1999. Laccases: a useful group of oxidoreductive enzymes. Bioremediat. J. 3: 1-26.
- Gochev VK, Krastanov AI. 2007. Isolation of laccase producing Trichoderma spp. Bulg. J. Agric. Sci. 13: 171-176.
- Gomes SA, Rebelo MJ. 2003. A new laccase biosensor for polyphenols determination. Sensors 3: 166-175.
- Holker U, Dohse J, Hofer M. 2002. Extracellular laccases in ascomycetes Trichoderma atroviride and Trichoderma harzianum. Folia Microbiol. 47: 423-427.
- Jing DB, Li PJ, Stagnitti F, Xiong XZ. 2007. Optimization of laccase production from Trametes versicolor by solid fermentation. Can. J. Microbiol. 53: 245-251.
- Kantelinen A, Hatakka A, Viikari L. 1989. Production of lignin peroxidase and laccase by Phlebia radiata. Appl. Microbiol. Biotechnol. 31: 234-239.
- Lee IY, Jung KH, Lee CH, Park YH. 1999. Enhanced production of laccase in Trametes versicolor by the addition of ethanol. Biotechnol. Lett. 21: 965-968.
- Nemec T, Jernejc K. 2002. Influence of Tween 80 on lipid metabolism of an Aspergillus niger strain. Appl. Biochem. Biotechnol. 101: 229-238.
- Nyanhongo GS, Gomes J, Gubitz GM, Zvauya R, Read JS, Steiner W. 2002. Decolorization of textile dyes by laccases from a newly isolated strain of Trametes modesta. Water Res. 36: 1449-1456.
- Nyanhongo G, Gomes J, Cubitz G, Zvauya R, Read J, Steiner W. 2002. Production of laccase by a newly isolated strain of Trametes modesta. Bioresour. Technol. 84: 259-263.
- Palmieri G, Giardina P, Bianco C, Fontanella B, Sannia G. 2000. Copper induction of laccase isoenzymes in the ligninolytic fungus Pleurotus ostreatus. Appl. Environ. Microbiol. 66: 920-924.
- Paramjit KB, Daljit SA. 2009. Comparative production of ligninolytic enzymes by Phanerochaete chrysosporium and Polyporus sanguineus. Can. J. Microbiol. 55: 1397-1402.
- Patel H, Gupte A, Gupte S. 2009. Effect of different cultural conditions and inducers on production of laccase by a basidiomycete fungal isolate Pleurotus ostreatus HP-1 under solid state fermentation. Bioresources 4: 268-284.
- Pointing SB, Pelling AL, Smith GJD, Hyde KD, Reddy CA. 2005. Screening of basidiomycetes and xylariaceous fungi for lignin peroxidase and laccase gene-specific sequences. Mycol. Res. 109: 115-124.
- Pointing SB, Jones EBG, Vrijmoed LLP. 2000. Optimization of laccase production by Pycnoporus sanguineus in submerged liquid culture. Mycologia 92: 139-144.
- Power T, Ortoneda M, Morrissey JP, Dobson ADW. 2006. Differential expression of genes involved in iron metabolism in Aspergillus fumigatus. Int. Microbiol. 9: 281-287.
- Rodríguez A, Falcon MA, Carnicero A, Perestelo F, De la Fuente G, Trojanowski J. 1996. Laccase activities of Penicillium chrysogenum in relation to lignin degradation. Appl. Microbiol. Biotechnol. 45: 399–403.
- Savoie JM, Mata G, Billette C. 1998. Extracellular laccase production during hyphal interactions between Trichoderma sp. and Shiitake, Lentinula edodes. Appl. Microbiol. Biotechnol. 49: 589-593.
- Sethuraman A, Akin DE, Erriksson KE. 1999. Production of ligninolytic enzymes and synthetic lignin mineralization by the bird’s nest fungus Cyathus stercoreus. Appl. Mcirobiol. Biotechnol. 52: 689-697.
- Sharma KK, Kapoor M, Kuhad RC. 2005. In vivo enzymatic digestion, in vitro xylanase digestion, metabolic analogues, surfactants and polyethylene glycol ameliorate laccase production from Ganoderma sp. kk-02. Lett. Appl. Microbiol. 41: 24-31.
- Soden DM, Dobson ADW. 2001. Differential regulation of laccase gene expression in Pleurotus sajor-caju. Microbiology 147: 1755-1763.
- Stajic M, Persky L, Friesem D, Hadar Y, Wasser SP, Nevo E, et al. 2006. Effect of different carbon and nitrogen sources on laccase and peroxidases production by selected Pleurotus species. Enzyme Microb. Technol. 38: 65-73.
- Vasconcelos AFD, Barbosa AM, Dekker RFH, Scarmínio IS, Rezende MI. 2000. Optimization of laccase production by Botryosphaeria sp. in the presence of veratryl alcohol by the response-surface method. Process Biochem. 35: 1131-1138.
- Velazquez-Cedeiio MA, Farnet AM, Ferre E, Savoie JM. 2004. Variations of lignocellulosic activities in dual cultures of Pleurotus ostreatus and Trichoderma longibrachiatum on unsterilized wheat straw. Mycologia 96: 712-719.
- Warsinke A, Benkert A, Scheller FW. 2000. Electrochemical immunoassays. Fresen. J. Anal. Chem. 366: 622-634.
- Zhang SX, Xiao YZ, Wang YP. 2004. Immobilization of fungal laccase on nylon net and application of the immobilized enzyme. Microbiology 31: 85-88.
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Research article
J. Microbiol. Biotechnol. 2013; 23(12): 1757-1764
Published online December 28, 2013 https://doi.org/10.4014/jmb.1302.02057
Copyright © The Korean Society for Microbiology and Biotechnology.
Media Optimization for Laccase Production by Trichoderma harzianum ZF-2 Using Response Surface Methodology
Huiju Gao 1, Xiang Chu 1, Yanwen Wang 1*, Fei Zhou 1, Kai Zhao 1, Zhimei Mu 1 and Qingxin Liu 1
College of Forestry, Shandong Agricultural University, Shandong 271018, China
Received: February 25, 2013; Accepted: September 12, 2013
Abstract
Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from
Shandong, China, and showed dye decolorization activities. The objective of this study was to
optimize its culture conditions using a statistical analysis of its laccase production. The
interactions between different fermentation parameters for laccase production were
characterized using a Plackett-Burman design and the response surface methodology. The
different media components were initially optimized using the conventional one-factor-at-atime
method and an orthogonal test design, and a Plackett-Burman experiment was then
performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal,
and CuSO4 were all found to have a significant influence on laccase production, and the
optimal concentrations of these three factors were then sequentially investigated using the
response surface methodology with a central composite design. The resulting optimal medium
components for laccase production were determined as follows: wheat straw powder 7.63 g/l,
soybean meal 23.07 g/l, (NH4)2SO4 1 g/l, CuSO4 0.51 g/l, Tween-20 1 g/l, MgSO4 1 g/l, and
KH2PO4 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased
59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized
medium. This is the first report on the statistical optimization of laccase production by
Trichoderma harzianum ZF-2.
Keywords: Trichoderma harzianum ZF-2, media optimization, laccase production, response surface methodology
References
- Abadulla E, Tzanov T, Costa S, Robra KH, Cavaco PA, Gubitz G. 2000. Decolorization and detoxication of textile dyes with a laccase from Trametes hirsuta. Appl. Environ. Microbiol. 66: 3357-3362.
- Assavanig A, Amornkitticharoen B, Ekpaisal N, Meevootisom V, Flegel TM. 1992. Isolation, characterization and function of laccase from Trichoderma. Appl. Microbiol. Biotechnol. 38:198-202.
- Bollag JM, Leonowicz A. 1984. Comparative studies of extracellular fungal laccases. Appl. Environ. Microbiol. 48:849-854.
- Box GEP, Wilson KB. 1951. On the experimental attainment of optimum conditions. J. Roy. Stat. Soc. B 13: 145.
- Chen T, Barton SC, Binyamin G, Gao Z, Zhang Y, Kim HH, et al. 2001. A miniature biofuel cell. J. Am. Chem. Soc. 123:8630-8631.
- Collins PJ, Dobson ADW. 1997. Regulation of laccase gene transcription in Trametes versicolor. Appl. Environ. Microbiol. 63: 3444-3450.
- Dittmer JK, Patel NJ, Dhawale SW, Dhawale SS. 1997. Production of multiple laccase isoforms by Phanerochaete chrysosporium grown under nutrient sufficiency. FEMS Microbiol. Lett. 149: 65-70.
- Eggert C, Temp U, Eriksson KEL. 1996. The ligninolytic system of the white-rot fungus Pycnoporus cinnabarinus: purification and characterization of the laccase. Appl. Environ. Microbiol. 62: 1151-1158.
- Gao HJ, Wang YW, Zhang WT, Wang WL, Mu ZM. 2011. Isolation, identification and application in lignin degradation of an ascomycete GHJ-4. Afr. J. Biotechnol. 10: 4166-4174.
- Gianfreda L, Xu F, Bollag JM. 1999. Laccases: a useful group of oxidoreductive enzymes. Bioremediat. J. 3: 1-26.
- Gochev VK, Krastanov AI. 2007. Isolation of laccase producing Trichoderma spp. Bulg. J. Agric. Sci. 13: 171-176.
- Gomes SA, Rebelo MJ. 2003. A new laccase biosensor for polyphenols determination. Sensors 3: 166-175.
- Holker U, Dohse J, Hofer M. 2002. Extracellular laccases in ascomycetes Trichoderma atroviride and Trichoderma harzianum. Folia Microbiol. 47: 423-427.
- Jing DB, Li PJ, Stagnitti F, Xiong XZ. 2007. Optimization of laccase production from Trametes versicolor by solid fermentation. Can. J. Microbiol. 53: 245-251.
- Kantelinen A, Hatakka A, Viikari L. 1989. Production of lignin peroxidase and laccase by Phlebia radiata. Appl. Microbiol. Biotechnol. 31: 234-239.
- Lee IY, Jung KH, Lee CH, Park YH. 1999. Enhanced production of laccase in Trametes versicolor by the addition of ethanol. Biotechnol. Lett. 21: 965-968.
- Nemec T, Jernejc K. 2002. Influence of Tween 80 on lipid metabolism of an Aspergillus niger strain. Appl. Biochem. Biotechnol. 101: 229-238.
- Nyanhongo GS, Gomes J, Gubitz GM, Zvauya R, Read JS, Steiner W. 2002. Decolorization of textile dyes by laccases from a newly isolated strain of Trametes modesta. Water Res. 36: 1449-1456.
- Nyanhongo G, Gomes J, Cubitz G, Zvauya R, Read J, Steiner W. 2002. Production of laccase by a newly isolated strain of Trametes modesta. Bioresour. Technol. 84: 259-263.
- Palmieri G, Giardina P, Bianco C, Fontanella B, Sannia G. 2000. Copper induction of laccase isoenzymes in the ligninolytic fungus Pleurotus ostreatus. Appl. Environ. Microbiol. 66: 920-924.
- Paramjit KB, Daljit SA. 2009. Comparative production of ligninolytic enzymes by Phanerochaete chrysosporium and Polyporus sanguineus. Can. J. Microbiol. 55: 1397-1402.
- Patel H, Gupte A, Gupte S. 2009. Effect of different cultural conditions and inducers on production of laccase by a basidiomycete fungal isolate Pleurotus ostreatus HP-1 under solid state fermentation. Bioresources 4: 268-284.
- Pointing SB, Pelling AL, Smith GJD, Hyde KD, Reddy CA. 2005. Screening of basidiomycetes and xylariaceous fungi for lignin peroxidase and laccase gene-specific sequences. Mycol. Res. 109: 115-124.
- Pointing SB, Jones EBG, Vrijmoed LLP. 2000. Optimization of laccase production by Pycnoporus sanguineus in submerged liquid culture. Mycologia 92: 139-144.
- Power T, Ortoneda M, Morrissey JP, Dobson ADW. 2006. Differential expression of genes involved in iron metabolism in Aspergillus fumigatus. Int. Microbiol. 9: 281-287.
- Rodríguez A, Falcon MA, Carnicero A, Perestelo F, De la Fuente G, Trojanowski J. 1996. Laccase activities of Penicillium chrysogenum in relation to lignin degradation. Appl. Microbiol. Biotechnol. 45: 399–403.
- Savoie JM, Mata G, Billette C. 1998. Extracellular laccase production during hyphal interactions between Trichoderma sp. and Shiitake, Lentinula edodes. Appl. Microbiol. Biotechnol. 49: 589-593.
- Sethuraman A, Akin DE, Erriksson KE. 1999. Production of ligninolytic enzymes and synthetic lignin mineralization by the bird’s nest fungus Cyathus stercoreus. Appl. Mcirobiol. Biotechnol. 52: 689-697.
- Sharma KK, Kapoor M, Kuhad RC. 2005. In vivo enzymatic digestion, in vitro xylanase digestion, metabolic analogues, surfactants and polyethylene glycol ameliorate laccase production from Ganoderma sp. kk-02. Lett. Appl. Microbiol. 41: 24-31.
- Soden DM, Dobson ADW. 2001. Differential regulation of laccase gene expression in Pleurotus sajor-caju. Microbiology 147: 1755-1763.
- Stajic M, Persky L, Friesem D, Hadar Y, Wasser SP, Nevo E, et al. 2006. Effect of different carbon and nitrogen sources on laccase and peroxidases production by selected Pleurotus species. Enzyme Microb. Technol. 38: 65-73.
- Vasconcelos AFD, Barbosa AM, Dekker RFH, Scarmínio IS, Rezende MI. 2000. Optimization of laccase production by Botryosphaeria sp. in the presence of veratryl alcohol by the response-surface method. Process Biochem. 35: 1131-1138.
- Velazquez-Cedeiio MA, Farnet AM, Ferre E, Savoie JM. 2004. Variations of lignocellulosic activities in dual cultures of Pleurotus ostreatus and Trichoderma longibrachiatum on unsterilized wheat straw. Mycologia 96: 712-719.
- Warsinke A, Benkert A, Scheller FW. 2000. Electrochemical immunoassays. Fresen. J. Anal. Chem. 366: 622-634.
- Zhang SX, Xiao YZ, Wang YP. 2004. Immobilization of fungal laccase on nylon net and application of the immobilized enzyme. Microbiology 31: 85-88.
