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

Babič J, Likozar B, Pavko A - Int J Mol Sci (2012)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Chair of Chemical, Biochemical and Environmental Engineering, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, 1000 Ljubljana, Slovenia; E-Mail: janja.babic@fkkt.uni-lj.si.

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.

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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).
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f2-ijms-13-11365: 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).

Mentions: In the production of enzymes, especially in large-scale processing, antifoaming agents and surfactants are normally used. Polysorbate 80 (Tween 80) is a non-ionic surfactant that is helpful in releasing fungal enzymes to the external environment. Although Polysorbate 80 is of low toxicity to the cellular membrane, it can alter the structure and morphology of fungi and the bacterial cell wall, leading to an increase of protein secretion [26]. Figure 2 shows the influence of Polysorbate 80 on the activity of Lac and MnP for the fungus C. subvermispora in a shaken flask experiment. The initial concentrations of nitrogen and carbon sources were 5.50 g L−1 for glucose and 1.05 g L−1 for ammonium tartrate. Lac and MnP activities in the filtrate are 10 times higher in the presence of a low concentration of Polysorbate 80 (0.1 g L−1). The results thus showed that Polysorbate 80 could be used as a stimulatory agent in submerged fungal cultivation, but in low concentrations. Figure 1b,c,e,f,h,i (model predictions) also show that either at fixed glucose (x1 = 0.5, CG = 5.50 g L−1) or ammonium tartrate concentrations (x2 = 0.5, CDT = 1.05 g L−1), the concentration of Polysorbate 80 (x3) should be as low as possible. The dependence on nitrogen or carbon source exhibits a maximum. The latter is again within medium levels of glucose (2–6 g L−1) and ammonium tartrate (0.8–1.2 g L−1).


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

Babič J, Likozar B, Pavko A - Int J Mol Sci (2012)

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).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472751&req=5

f2-ijms-13-11365: 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).
Mentions: In the production of enzymes, especially in large-scale processing, antifoaming agents and surfactants are normally used. Polysorbate 80 (Tween 80) is a non-ionic surfactant that is helpful in releasing fungal enzymes to the external environment. Although Polysorbate 80 is of low toxicity to the cellular membrane, it can alter the structure and morphology of fungi and the bacterial cell wall, leading to an increase of protein secretion [26]. Figure 2 shows the influence of Polysorbate 80 on the activity of Lac and MnP for the fungus C. subvermispora in a shaken flask experiment. The initial concentrations of nitrogen and carbon sources were 5.50 g L−1 for glucose and 1.05 g L−1 for ammonium tartrate. Lac and MnP activities in the filtrate are 10 times higher in the presence of a low concentration of Polysorbate 80 (0.1 g L−1). The results thus showed that Polysorbate 80 could be used as a stimulatory agent in submerged fungal cultivation, but in low concentrations. Figure 1b,c,e,f,h,i (model predictions) also show that either at fixed glucose (x1 = 0.5, CG = 5.50 g L−1) or ammonium tartrate concentrations (x2 = 0.5, CDT = 1.05 g L−1), the concentration of Polysorbate 80 (x3) should be as low as possible. The dependence on nitrogen or carbon source exhibits a maximum. The latter is again within medium levels of glucose (2–6 g L−1) and ammonium tartrate (0.8–1.2 g L−1).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Chair of Chemical, Biochemical and Environmental Engineering, Faculty of Chemistry and Chemical Technology, University of Ljubljana, Aškerčeva 5, 1000 Ljubljana, Slovenia; E-Mail: janja.babic@fkkt.uni-lj.si.

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.

Show MeSH