LACCASE ISOLATION FROM SPENT MUSHROOM WASTE (AGARICUS BISPORUS): PARTIAL PURIFICATION, CHARACTERIZATION, IMMOBILIZATION AND BIOTECHNOLOGICAL APPLICATION

Year 2021, , 44 - 56, 25.01.2021

Güliz Ak , Habibe Yılmaz , Şenay Şanlıer

https://doi.org/10.18036/estubtdc.726398

Abstract

Laccase was partially purified from mushroom compost with cheap and simple purification methods. Partially purified laccase was immobilized onto Amberlite XAD-7 resin. Immobilization conditions and textile dye (Direct Green B) decolorization of immobilized enzyme were examined. Laccase was partially purified from mushroom compost with 3.22 purification fold. Immobilization time for laccase was 30 minutes with 97% efficiency. The immobilization yield based on the activity was calculated by about 90%. The immobilization yield was calculated as nearly 95% based on the measurement of protein amount at each stage. Immobilized enzyme preserved its initial activity with an 84% rate even after its tenth use. The free enzyme lost its activity immediately when it was incubated with the dye solution under its optimal conditions (pH 3.0 and 65°C). On the contrary, the immobilized enzyme maintained its initial activity with a 53% rate when incubated with dye solution under its optimal conditions (pH 3.5 and 65°C).

Keywords

Mushroom compost, Laccase, Direct Green B, Diamine Green B, Enzyme Immobilization, Dye decolorization, Amberlite XAD-7

Supporting Institution

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Project Number

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Thanks

The authors acknowledge to Prof. Dr. İhsan YASA for gifting the mushroom compost. The authors have declared no conflict of interest.

LACCASE ISOLATION FROM SPENT MUSHROOM WASTE (AGARICUS BISPORUS): PARTIAL PURIFICATION, CHARACTERIZATION, IMMOBILIZATION AND BIOTECHNOLOGICAL APPLICATION

Abstract

Lakkaz, ucuz ve basit saflaştırma yöntemleriyle mantar kompostundan kısmen saflaştırılmıştır. Kısmen saflaştırılmış lakkaz, Amberlite XAD-7 reçinesi üzerine immobilize edilmiştir. İmmobilizasyon koşulları ve immobilize enzimin tekstil boyasının (Direct Green B) renk giderimi incelenmiştir. Lakkaz, mantar kompostundan 3,22 saflaştırma katı ile kısmen saflaştırılmıştır. Lakkaz için immobilizasyon süresi % 97 verimlilikle 30 dakikadır. Aktiviteye dayanan immobilizasyon verimi yaklaşık % 90 olarak hesaplanmıştır. İmmobilizasyon verimi, her aşamadaki protein miktarının ölçülmesine dayanılarak yaklaşık % 95 olarak hesaplanmıştır. İmmobilize enzim onuncu kullanımdan sonra bile başlangıçtaki aktivitesini % 84 oranında korumuştur. Serbest enzim, optimal koşullar (pH 3,0 ve 65°C) altında boya çözeltisi ile inkübe edildiğinde hemen aktivitesini kaybetmiştir. Aksine, immobilize enzim, optimal koşullar (pH 3,5 ve 65°C) altında boya çözeltisi ile inkübe edildiğinde başlangıç aktivitesini % 53 oranında sürdürmüştür.

Keywords

Mantar kompostu, Lakkaz, Direct Green B, Diamine Green B, Enzim immobilizasyonu, Boya renk giderimi, Amberlite XAD-7

Project Number

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References

• Chang ST & Miles PG. A New Look at Cultivated Mushrooms. Bioscience. 1984; 34(6): 358-362.
• Phillips R & Reid DR. Mushrooms, UK. London: Pan Macmillan Limited, 2006.
• Alhulajaily A, Yu H, Zhang X, Ma F. Highly Efficient and Sustainable Spent Mushroom Waste Adsorbent Based on Surfactant Modification for the Removal of Toxic Dyes. Int. J. Environ. Res. Public Health. 2018; 15: 1421-1436.
• Lim SH, Lee YH, Kang HW. Efficient Recovery of Lignocellulolytic Enzymes of Spent Mushroom Compost from Oyster Mushrooms, Pleurotus spp., and Potential Use in Dye Decolorization. Mycobiology. 2013; 41(4): 214-220.
• Bilal M and Asgher M. Biodegradation of agrowastes by lignocellulolytic activity of an oyster mushroom, Pleurotus sapidus. J. Natn. Sci. Foundation Sri Lanka. 2016; 44(4): 399-407.
• Oliveira SF, Rodrigues da Luz JM, Kasuya MCM, Ladeira LO, Junior AC. Enzymatic extract containing lignin peroxidase immobilized on carbon nanotubes: Potential biocatalyst in dye decolourization. Saudi J. Biol. Sci.. 2018; 25(4): 651-659.
• Machado KMG and Matheus DR. Biodegradation of remazol brilliant blue R by ligninolytic enzymatic complex produced by Pleurotus ostreatus. Braz. J. Microbiol. 2006; 37(4): 468-473.
• Mayer AM & Staples RC. Laccase: new functions for an old enzyme. Phytochemistry. 2002; 60(6): 551-565.
• Claus H. Laccases: structure, reactions, distribution. Micron. 2004; 35(1-2): 93-96.
• Madhavi, V., & Lele, S. S. 2009. Laccase: Properties and applications. BioResources, 4(4), 1694-1717.
• Chhabra M, Mishra S, Sreekrishnan TR. Immobilized laccase mediated dye decolorization and transformation pathway of azo dye acid red 27. J Environ Health Sci Eng. 2015; 13(38): 1-9.
• Couto SR & Toca-Herrera JL. Lacasses in the textile industry. Biotechnol. Mol. Biol. Rev. 2006; 1(4): 115-120.
• Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976; 72: 248-254.
• Hamarat Sanlier Ş, Gider S, Köprülü S. Immobilization of laccase for biotechnology applications. Artif. Cells Nanomed. Biotechnol. 2012; 41(4): 259-263.
• Pezella C, Russo ME, Marzocchella A, Salatino P, Sannia G. Immobilization of a Pleurotus ostreatus Laccase Mixture on Perlite and Its Application to Dye Decolourisation. Biomed Res. Int. 2014: 308613.
• Liu Y, Zeng Z, Zeng G, Tang L, Pang Y, Li Z, Liu C, Lei X, Wu M, Ren P, Liu Z, Chen M, Xie G. Immobilization of Laccase on Magnetic Bimodal Mesoporous Carbon and the Application in the Removal of Phenolic Compounds. Bioresour. Technol. 2012; 115: 21-26.
• Lassouane F, Aït-Amar H, Amrani S, Rodriguez-Couto S. A promising laccase immobilization approach for Bisphenol A removal from aqueous solutions. Bioresour. Technol. 2019; 271: 360-367.
• Xu R, Tang R, Zhou Q, Li F, Zhang B. Enhancement of catalytic activity of immobilized laccase for diclofenac biodegradation by carbon nanotubes. Chem Eng J. 2015; 262: 88-95.
• Wong P, Fan X, Cui L, Wong Q, Zhan A. Decolorization of reactive dyes by laccase immobilized in alginate/gelatin blent with PEG. J. Envıron. Scı-Chına. 2008; 20: 1519-1522.
• Bayramoglu G, Yilmaz M, Arica YM. Reversible immobilization of laccase to poly(4-vinylpyridine) grafted and Cu(II) chelated magnetic beads: Biodegradation of reactive dyes. Bioresour. Technol. 2010; 101: 6615-6621.
• Cristovao RO, Tavares APM, Brigida AI, Loureiro JM, Boaventura RAR, Macedo EA, Coelho MAZ. Immobilization of commercial laccase onto green coconut fiber by adsorption and its application for reactive textile dyes degradation. J. Mol. Catal., B Enzym. 2011; 72: 6-12.