Ligninolytic enzymes from white-rot fungi are widely used in biotechnological processes. However, the application of these enzymes as free enzymes is limited due to their instability and lack of reusability. Enzyme stabilization is therefore a major challenge in biocatalytic process research, and immobilization methods are desirable. Using cross-linked enzyme aggregates (CLEAs) such as magnetic CLEAs, porous-CLEAs and combi-CLEAs is a promising technique for overcoming these issues. Cross-linking methods can stabilize and immobilize enzymes by interconnecting enzyme molecules via multiple bonds using cross-linking agents such as glutaraldehyde. The high catalyst density and microporous assembly of CLEAs guarantee high catalyst activity, which, together with their long shelf life, operational stability, and reusability, provide a cost-efficient alternative to matrix-assisted immobilization approaches. Here, we review current progress in ligninolytic enzyme immobilization and provide a comprehensive review of CLEAs. Moreover, we summarize the use of these CLEAs for biocatalysis processes, bioremediation such as dye decolourization, wastewater treatment or pharmaceutically active compound elimination.
The key to obtaining an optimum performance of an enzyme is often a question of devising a suitable enzyme and optimisation of conditions for its immobilization. In this study, laccases from the native isolates of white rot fungi Fomes fomentarius and/or Trametes versicolor, obtained from Czech forests, were used. From these, cross-linked enzyme aggregates (CLEA) were prepared and characterised when the experimental conditions were optimized. Based on the optimization steps, saturated ammonium sulphate solution (75 wt.%) was used as the precipitating agent, and different concentrations of glutaraldehyde as a cross-linking agent were investigated. CLEA aggregates formed under the optimal conditions showed higher catalytic efficiency and stabilities (thermal, pH, and storage, against denaturation) as well as high reusability compared to free laccase for both fungal strains. The best concentration of glutaraldehyde seemed to be 50 mM and higher efficiency of cross-linking was observed at a low temperature 4 °C. An insignificant increase in optimum pH for CLEA laccases with respect to free laccases for both fungi was observed. The results show that the optimum temperature for both free laccase and CLEA laccase was 35 °C for T. versicolor and 30 °C for F. fomentarius. The CLEAs retained 80% of their initial activity for Trametes and 74% for Fomes after 70 days of cultivation. Prepared cross-linked enzyme aggregates were also investigated for their decolourisation activity on malachite green, bromothymol blue, and methyl red dyes. Immobilised CLEA laccase from Trametes versicolor showed 95% decolourisation potential and CLEA from Fomes fomentarius demonstrated 90% decolourisation efficiency within 10 h for all dyes used. These results suggest that these CLEAs have promising potential in dye decolourisation.
- MeSH
- azosloučeniny chemie MeSH
- barva MeSH
- barvicí látky chemie MeSH
- bromthymolová modř chemie MeSH
- enzymy imobilizované chemie MeSH
- glutaraldehyd chemie MeSH
- katalýza MeSH
- lakasa chemie MeSH
- Polyporales enzymologie MeSH
- reagencia zkříženě vázaná chemie MeSH
- rosanilinová barviva chemie MeSH
- síran amonný chemie MeSH
- teplota MeSH
- Trametes enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
An investigation was carried out on the effect of inoculation methods on the compost of an organic fraction of municipal solid waste. Three types of white-rot fungi (Phanerochaete chrysosporium, Trametes versicolor and Fomes fomentarius), and a consortium of these fungi, were used. The study assessed their influence on microbial enzymatic activities and the quality of the finished compost. It was found that the addition of white-rot fungi to municipal solid waste (after 37days of composting) could be a useful strategy for enhancing the properties of the final compost product. In comparison with the control sample (compost without inoculation), it accelerates degradation of solid waste as indicated by changes in C/N, electrical conductivity and pH. However, the effectiveness of waste degradation and compost maturation depends on the type of microorganism used for inoculation. The presence of inoculants, such as Trametes versicolor and Fomes fomentarius, led to a higher degrading ratio and a better degree of maturity. This resulted in an increase of enzymatic activities (especially dehydrogenase and protease) and a germination index in comparison with inoculation using Phanerochaete chrysosporium or a consortium of fungi.
- MeSH
- biodegradace MeSH
- Coriolaceae metabolismus MeSH
- dusík metabolismus MeSH
- enzymy metabolismus MeSH
- houby metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- odpadky - odstraňování metody MeSH
- Phanerochaete metabolismus MeSH
- půda * MeSH
- půdní mikrobiologie * MeSH
- Trametes metabolismus MeSH
- tuhý odpad * MeSH
- uhlík metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Ligninolytic enzymes, such as laccase, lignin peroxidase and manganese peroxidase, are biotechnologically-important enzymes. The ability of five white-rot fungal strains Daedaleopsis confragosa, Fomes fomentarius, Trametes gibbosa, Trametes suaveolens and Trametes versicolor to produce these enzymes has been studied. Three different copper(II) complexes have been prepared ((Him)[Cu(im)₄(H₂O)₂](btc)·3H₂O, where im = imidazole, H₃btc = 1,3,5-benzenetricarboxylic acid, [Cu₃(pmdien)₃(btc)](ClO₄)₃·6H₂O) and [Cu₃(mdpta)₃(btc)](ClO₄)₃·4H₂O, where pmdien = N,N,N',N'',N''-pentamethyl-diethylenetriamine and mdpta = N,N-bis-(3-aminopropyl)methyl- amine), and their potential application for laccase and peroxidases induction have been tested. The enzyme-inducing activities of the complexes were compared with that of copper sulfate, and it has been found that all of the complexes are suitable for the induction of laccase and peroxidase activities in white-rot fungi; however, the newly-synthesized complex M1 showed the greatest potential for the induction. With respect to the different copper inducers, this parameter seems to be important for enzyme activity, which depends also on the fungal strains.
- MeSH
- enzymová indukce MeSH
- fungální proteiny biosyntéza genetika MeSH
- komplexní sloučeniny chemie farmakologie MeSH
- lakasa biosyntéza genetika MeSH
- měď chemie MeSH
- peroxidasy biosyntéza genetika MeSH
- regulace genové exprese enzymů MeSH
- Trametes enzymologie MeSH
- vodíková vazba MeSH
- Publikační typ
- časopisecké články MeSH
