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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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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.
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f7-ijms-13-11365: 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.

Mentions: The second phase of the study continued with C. subvermispora cultivation experiments on various immobilization supports using the optimized liquid medium (OLM). Beech wood cubes (BWCM) and pine wood cubes (PWCM) containing natural lignin as inducer and inert polyurethane foam cubes (PUFCM) were used to immobilize the culture growth and to control the immobilization effect in comparison to the combined effect of the other supports that act as carriers as well as inducers. The composition of the liquid media was prepared according to the optimum settings for OFLac/MnP at w = 0.5 obtained from the response surface methodology experiment. In addition, the OLM was used as a reference medium with free mycelium. The results are presented in Figure 7. The maximal enzyme activities with free mycelium in the OLM without immobilization support were achieved after eight days, namely 108 U L−1 for MnP and 19 U L−1 for Lac. It is evident that the use of wood supports affected the enzyme activities. Beech wood and pine wood induced higher Lac activities. Maximal values using BWCM and PWCM, achieved after eight days of cultivation, were 150 U L−1 and 135 U L−1, respectively. In contrast, MnP activities using wood supports were lower than the results with only liquid medium OLM, namely 60 U L−1 for BWCM and 20 U L−1 for PWCM. Production of enzymes was the lowest under PUFCM cultivation conditions: MnP activity was only 40 U L−1, while Lac activity was close to zero (2 U L−1). In the literature it was reported that inert materials such as PUF affect fungal growth as an immobilization surface, but alone cannot increase enzyme synthesis [16,20]. In contrast, natural materials such as straw or wood with structural cell-wall components like cellulose, hemicelluloses and lignin form carbon sources available for fungal growth. In addition, wood contains small amounts of soluble sugars, lipids, peptides and starch as well as minerals and a wide range of extractives and volatiles. All these compounds vary with tree species like pine, beech or oak and therefore affect fungal growth and enzyme production in different ways [27]. Hence, they can be utilized both as an immobilization surface and a natural inducer [20,21]. Present results are also in agreement with our previous investigation, where the synthesis of ‘tailor made’ enzyme composition, that is excess Lac or excess MnP, was proposed by variation of the carbon and nitrogen source in the medium and the use of immobilization supports of different types of wood [21].


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)

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

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

f7-ijms-13-11365: 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.
Mentions: The second phase of the study continued with C. subvermispora cultivation experiments on various immobilization supports using the optimized liquid medium (OLM). Beech wood cubes (BWCM) and pine wood cubes (PWCM) containing natural lignin as inducer and inert polyurethane foam cubes (PUFCM) were used to immobilize the culture growth and to control the immobilization effect in comparison to the combined effect of the other supports that act as carriers as well as inducers. The composition of the liquid media was prepared according to the optimum settings for OFLac/MnP at w = 0.5 obtained from the response surface methodology experiment. In addition, the OLM was used as a reference medium with free mycelium. The results are presented in Figure 7. The maximal enzyme activities with free mycelium in the OLM without immobilization support were achieved after eight days, namely 108 U L−1 for MnP and 19 U L−1 for Lac. It is evident that the use of wood supports affected the enzyme activities. Beech wood and pine wood induced higher Lac activities. Maximal values using BWCM and PWCM, achieved after eight days of cultivation, were 150 U L−1 and 135 U L−1, respectively. In contrast, MnP activities using wood supports were lower than the results with only liquid medium OLM, namely 60 U L−1 for BWCM and 20 U L−1 for PWCM. Production of enzymes was the lowest under PUFCM cultivation conditions: MnP activity was only 40 U L−1, while Lac activity was close to zero (2 U L−1). In the literature it was reported that inert materials such as PUF affect fungal growth as an immobilization surface, but alone cannot increase enzyme synthesis [16,20]. In contrast, natural materials such as straw or wood with structural cell-wall components like cellulose, hemicelluloses and lignin form carbon sources available for fungal growth. In addition, wood contains small amounts of soluble sugars, lipids, peptides and starch as well as minerals and a wide range of extractives and volatiles. All these compounds vary with tree species like pine, beech or oak and therefore affect fungal growth and enzyme production in different ways [27]. Hence, they can be utilized both as an immobilization surface and a natural inducer [20,21]. Present results are also in agreement with our previous investigation, where the synthesis of ‘tailor made’ enzyme composition, that is excess Lac or excess MnP, was proposed by variation of the carbon and nitrogen source in the medium and the use of immobilization supports of different types of wood [21].

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