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. 2012;13(9):11365-11384.
doi: 10.3390/ijms130911365. Epub 2012 Sep 12.

Optimization of ligninolytic enzyme activity and production rate with Ceriporiopsis subvermispora for application in bioremediation by varying submerged media composition and growth immobilization support

Janja Babič  1 , Blaž Likozar  2 , Aleksander Pavko  1
Affiliations

Optimization of ligninolytic enzyme activity and production rate with Ceriporiopsis subvermispora for application in bioremediation by varying submerged media composition and growth immobilization support

Janja Babič et al. Int J Mol Sci. 2012.

Abstract

Response surface methodology (central composite design of experiments) was employed to simultaneously optimize enzyme production and productivities of two ligninolytic enzymes produced by Ceriporiopsis subvermispora. Concentrations of glucose, ammonium tartrate and Polysorbate 80 were varied to establish the optimal composition of liquid media (OLM), where the highest experimentally obtained activities and productivities were 41 U L(-1) and 16 U L(-1) day(-1) for laccase (Lac), and 193 U L(-1) and 80 U L(-1) day(-1) for manganese peroxidase (MnP). Considering culture growth in OLM on various types of immobilization support, the best results were obtained with 1 cm beech wood cubes (BWCM). Enzyme activities in culture filtrate were 152 U L(-1) for Lac and 58 U L(-1) for MnP, since the chemical composition of this immobilization material induced higher Lac activity. Lower enzyme activities were obtained with polyurethane foam. Culture filtrates of OLM and BWCM were applied for dye decolorization. Remazol Brilliant Blue R (RBBR) was decolorized faster and more efficiently than Copper(II)phthalocyanine (CuP) with BWCM (80% and 60%), since Lac played a crucial role. Decolorization of CuP was initially faster than that of RBBR, due to higher MnP activities in OLM. The extent of decolorization after 14 h was 60% for both dyes.

Keywords: bioremediation; decolorization; enzyme production; immobilized culture; laccase; ligninolytic enzymes; manganese peroxidase; medium optimization; response surface methodology; submerged cultivation.

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Figures

Figure 1
Figure 1
Estimated response surface for OFLac (the contribution of relative laccase (Lac) activity to OFLac (w) was set at (a–c) 0.25, (d–f) 0.50 and (g–i) 0.75), on varying the initial glucose (x1), ammonium tartrate (x2) and Polysorbate 80 (x3) concentrations for batch experiments at a temperature of 30 °C.
Figure 2
Figure 2
Comparison of laccase (Lac) and manganese peroxidase (MnP) activity obtained from enzyme assays with ABTS (Lac) and DMP (MnP) in batch experiments at a temperature of 30 °C, an initial glucose concentration of 5.50 g L−1, an initial ammonium tartrate concentration of 1.05 g L−1, and an initial Polysorbate 80 concentration (CP) of 1.00 g L−1 (lower curves) and 0.10 g L−1 (upper curves).
Figure 3
Figure 3
(a) Standardized Pareto chart for the maximal Lac activity, [ELac]; and (b) its maximal production rate, d[ELac]/dt for batch experiments at a temperature of 30 °C; x1 is the initial glucose concentration; x2 is the initial ammonium tartrate concentration and x3 is the initial Polysorbate 80 concentration. Dotted line represents the 95% confidence interval margin.
Figure 4
Figure 4
Graph of optimal normalized highest values of experimentally obtained Lac and MnP activities (y1,Lac and y1,MnP), optimal normalized highest values of experimentally obtained Lac production rate (y2,Lac) and optimal objective function (OFLac, and OFLac/MnP) plotted against the contribution of the maximal enzyme activity divided by its highest experimental value (w).
Figure 5
Figure 5
Normalized and absolute optimum settings and OFLac at w = 0.5 for batch experiments at a temperature of 30 °C; the maximal enzyme activity and productivity (OFLac) are to be obtained after 4 days.
Figure 6
Figure 6
Normalized and absolute optimum settings and OFLac/MnP at w = 0.5 for batch experiments at a temperature of 30 °C; the maximal enzyme activities (OFLac/MnP) are to be obtained after 4 days.
Figure 7
Figure 7
Maximal activities of C. subvermispora MnP (■) and Lac (□) in experiments at a temperature of 30 °C with the optimized liquid medium (OLM) and in the medium with beech wood cubes (BWCM), pine wood cubes (PWCM) and inert polyurethane cubes (PUFCM) as immobilization support; the maximal enzyme activities were obtained after eight days.
Figure 8
Figure 8
Decolourization of Remazol Brilliant Blue R (RBBR) dye with C. subvermispora enzyme mixture from OLM (●) and BWCM (○) at room temperature. Experimental error is in the range of ±4%.
Figure 9
Figure 9
Decolourization of Copper (II) phthalocyanine (CuP) dye with C. subvermispora enzyme mixture from OLM (●) and BWCM (○) at room temperature. Experimental error is in the range of ±4%.
Figure 10
Figure 10
Schematic diagram of the RSM procedure.
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References

    1. Martínez A.T., Speranza M., Ruiz-Dueñas F.J., Ferreira P., Camarero S., Guillén F., Martínez M.J., Gutiérrez A., del Río J.C. Biodegradation of lignocellulosics: Microbial, chemical, and enzymatic aspects of the fungal attack of lignin. Int. Microbiol. 2005;8:195–204. - PubMed
    1. Enoki M., Watanabe T., Nakagame S., Koller K., Messner K., Honda Y., Kuwahara M. Extracellular lipid peroxidation of selective white-rot fungus, Ceriporiopsis subvermispora. FEMS Microbiol. Lett. 1999;180:205–211. - PubMed
    1. Tanaka H., Koike K., Itakura S., Enoki A. Degradation of wood and enzyme production by Ceriporiopsis subvermispora. Enzym. Microb. Technol. 2009;45:384–390.
    1. Maciel M.J., Silva A.C., Ribeiro H.C.T. Industrial and biotechnological applications of ligninolytic enzymes of the basidiomycota: A review. Electron. J. Biotechnol. 2010;13 doi: 10.2225/vol13-issue6-fulltext-2. - DOI
    1. Kirk T.K., Cullen D. In: Environmentally Friendly Technologies for the Pulp and Paper Industry. Young R.A., Akhtar M., editors. Wiley; New York, NY, USA: 1998. pp. 273–308.

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