Production of Laccase Enzyme in Various Media by Funalia trogii and Trametes versicolor in Repeated Batch Process

Yıl 2017, Cilt: 43 Sayı: 1, 27 - 40, 28.04.2017

Emre Birhanlı Özfer Yeşilada

Öz

Laccase enzyme (EC
1.10.3.2) is a polyphenol oxidase which has four copper ions in the structure
and the best laccase-producing organisms are white rot fungi. Laccase enzyme
which has a rather broad substrate specifity can be used in many
biotechnological, industrial and environmental fields. However, laccase enzyme
should be produced at large amounts as it can be used in these fields.
Therefore, it is extremely important to investigate high levels of
laccase-producing organisms, the cost-effective and appropriate enzyme
production media and also methods. In accordance with this purpose Funalia
trogii ATCC 200800 and Trametes versicolor ATCC 200801 which have high
laccase production potential were used as enzyme producers. Distilled water,
Sabouraud Dextrose Broth, Malt Extract Broth and cheese whey media with copper
and without copper were used as enzyme production media. Repeated batch process
is used as laccase production method which we have found as an efficient method
in our previous studies. The highest laccase activities for F. trogii
and T. versicolor were obtained from Malt Extract Broth as 17.68 and
6.27 U mL^-1, respectively. In terms of the laccase production, the
efficiency order of copper added media for both fungus is Malt Extract Broth
> Sabouraud Dextrose Broth > distilled water. While the efficiency order
of copper free media for F. trogii is Malt Extract Broth > Sabouraud
Dextrose Broth > 25% cheese whey > distilled water > 50% cheese whey,
the efficiency order of copper free media for T. versicolor is Malt
Extract Broth > 25% cheese whey > Sabouraud Dextrose Broth > 50%
cheese whey > distilled water. As a result both fungus could produce laccase
in all media and laccase production highly increased in copper added media.

Anahtar Kelimeler

Funalia trogii, Trametes versicolor, Laccase, Copper

Tekrarlı Kesikli Süreçte Funalia trogii ve Trametes versicolor ile Lakkaz Enziminin Çeşitli Ortamlarda Üretimi

Öz

Lakkaz enzimi (EC 1.10.3.2) yapısında dört bakır iyonu bulunan bir polifenol oksidaz olup, en iyi lakkaz üreten organizmalar beyaz çürükçül funguslardır. Oldukça geniş bir substrat özgüllüğüne sahip olan lakkaz enzimi pek çok biyoteknolojik, endüstriyel ve çevresel alanlarda kullanılabilmektedir. Ancak lakkaz enziminin bu alanlarda kullanılabilmesi için yüksek miktarlarda üretilmesi gerekir. Bu nedenle yüksek miktarlarda lakkaz üreten organizmaların, düşük maliyetli ve uygun enzim üretim ortamlarının ve de yöntemlerinin araştırılması son derece önemlidir. Bu amaç doğrultusunda enzim üreticisi olarak yüksek lakkaz üretim potansiyeline sahip olan Funalia trogii ATCC 200800 ve Trametes versicolor ATCC 200801 kullanılmıştır. Enzim üretim ortamları olarak bakır içeren ve bakır içermeyen distile su, Sabouraud Dekstroz Broth, Malt Ekstrakt Broth ve peynir altı suyu ortamları kullanılmıştır. Lakkaz üretim yöntemi olarak daha önceki çalışmalarımızda verimli bir yöntem olarak saptadığımız tekrarlı kesikli işlem kullanılmıştır. En yüksek lakkaz aktiviteleri bakır ilave edilmiş Malt Ekstrakt Broth ortamlarında F. trogii ve T. versicolor için sırasıyla 17.68 ve 6.27 U mL-1 olarak elde edilmiştir. Lakkaz üretimi açısından bakır ilave edilmiş ortamların etkinlik sıralaması her iki fungus için de; Malt Ekstrakt Broth > Sabouraud Dekstroz Broth > distile su şeklindedir. Bakırsız ortamlardaki etkinlik sıralaması F. trogii için Malt Ekstrakt Broth > Sabouraud Dekstroz Broth > %25 peynir altı suyu > distile su > %50 peynir altı suyu iken, T. versicolor için; Malt Ekstrakt Broth > %25 peynir altı suyu > Sabouraud Dekstroz Broth > %50 peynir altı suyu > distile su şeklindedir. Sonuç olarak her iki fungus da tüm ortamlarda lakkaz üretebilmiş ve bakırlı ortamlarda lakkaz üretimi son derece artmıştır.

Anahtar Kelimeler

Funalia trogii, Trametes versicolor, Lakkaz, Bakır

Kaynakça

• Amutha C, Abhijit M (2015). Screening and isolation of laccase producers, determination of optimal condition for growth, laccase production and choose the best strain. Journal of Bioremediation & Biodegradation 6: 1–8.
• Arora DS, Gill PK (2000). Laccase production by some white rot fungi under different nutritional conditions. Bioresource Technology 73: 283–285.
• Baldrian P (2003). Interactions of heavy metals with white-rot fungi. Enzyme and Microbial Technology 32: 78–91.
• Birhanli E, Yesilada O (2006). Increased production of laccase by pellets of Funalia trogii ATCC 200800 and Trametes versicolor 200801 in repeated-batch mode. Enzyme and Microbial Technology 39: 1286–1293.
• Birhanli E, Yesilada O (2010). Enhanced production of laccase in repeated-batch cultures of Funalia trogii and Trametes versicolor. Biochemical Engineering Journal 52: 33–37.
• Birhanlı E, Yeşilada Ö (2013). The utilization of lignocellulosic wastes for laccase production under semisolid-state and submerged fermentation conditions. Turkish Journal of Biology 37: 450–456.
• Chandra R, Chowdhary P (2015). Properties of bacterial laccases and their application in bioremediation of industrial wastes. Environmental Science: Processes & Impacts 17: 326–342.
• Couto SR, Herrera JLT (2006). Industrial and biotechnological applications of laccases: A review. Biotechnology Advances 24: 500–513.
• Divya LM, Prasanth GK, Sadasivan C (2013). Isolation of a salt tolerant laccase secreting strain of Trichoderma sp. NFCCI–2745 and optimization of culture conditions and assessing its effectiveness in treating saline phenolic effluents. Journal of Environmental Sciences 25: 2410–2416.
• Janusz G, Rogalski J, Barwińska M, Szczodrak J (2006). Effects of culture conditions on production of extracellular laccase by Rhizoctonia praticola. Polish Journal of Microbiology 55: 309–319.
• Jegatheesan M, Eyini M (2015). Response surface methodology mediated modulation of laccase production by Polyporus arcularius. Arabian Journal for Science and Engineering 40: 1809–1818.
• Khan MMR, Ray M, Ray L, Guha AK (2016). Extracellular laccase from Pleurotus sajor-caju: Fermentative conditions and influence of nitrogenous sources. Indian Journal of Biotechnology 15: 230–235.
• Madhavi V, Lele SS (2009). Laccase: Properties and applications. Bioresources 4: 1694–1717.
• Manimozhi M, Kaviyarasan V (2012). Screening the effect of nutritional parameters on biomass and laccase production in submerged medium by litter decomposing basidiomycete Agaricus heterocystis. International Journal of Pharmacy and Pharmaceutical Sciences 4: 592–599.
• Mougin C, Kollmann A, Jolivalt C (2002). Enhanced production of laccase in the fungus Trametes versicolor by the addition of xenobiotics. Biotechnology Letters 24: 139–142.
• Otlu B (2002). Peyniraltı Suyu ve Alkol Fabrikası Atıksularının Arıtımı ve Değerlendirilmesi. Yüksek Lisans Tezi, İnönü Üniversitesi Fen Bilimleri Enstitüsü (Basılmamış), Malatya.
• Patel H, Gupte A (2016). Optimization of different culture conditions for enhanced laccase production and its purification from Tricholoma giganteum AGHP. Bioresources and Bioprocessing 3: 1–10.
• Pezzella C, Guarino L, Piscitelli A (2015). How to enjoy laccases. Cellular and Molecular Life Sciences 72: 923–940.
• Revankar MS, Lele SS (2006). Enhanced production of laccase using a new isolate of white rot fungus WR–1. Process Biochemistry 41: 581–588.
• Roth S, Spiess AC (2015). Laccases for biorefinery applications: A critical review on challenges and perspectives. Bioprocess and Biosystems Engineering 38: 2285–2313.
• Shutova VV, Revin VV, Myakushina YA (2008). The effect of copper ions on the production of laccase by the fungus Lentinus (Panus) tigrinus. Applied Biochemistry and Microbiology 44: 619–623.
• Singh AP, Singh T (2014). Biotechnological applications of wood-rotting fungi: A review. Biomass and Energy 62: 198–206.
• Siso MIG (1996). The biotechnological utilization of cheese whey: A review. Bioresource Technology 57: 1–11.
• Sun W, Xu M, Xia C, Li A, Sun G (2013). Enhanced production of laccase by Coriolus hirsutus using molasses distillery wastewater. Frontiers of Environmental Science & Engineering 7: 200–210.
• Vivekanand V, Dwivedi P, Pareek N, Singh RP (2011). Banana peel: A potential substrate for laccase production by Aspergillus fumigatus VkJ2.4.5 in solid-state fermentation. Applied Biochemistry and Biotechnology 165: 204–220.
• Yesilada O, Asma D, Cing S (2003). Decolorization of textile dyes by fungal pellets. Process Biochemistry 38: 933–938.
• Zhu C, Bao G, Huang S (2016). Optimization of laccase production in the white rot fungus Pleurotus ostreatus (ACCC 52857) induced through yeast extract and copper. Biotechnology and Biotechnological Equipment 30: 270–276.