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Production of Trametes pubescens laccase under submerged and semi-solid culture conditions on agro-industrial wastes.

Gonzalez JC, Medina SC, Rodriguez A, Osma JF, Alméciga-Díaz CJ, Sánchez OF - PLoS ONE (2013)

Bottom Line: The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2).Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3.In addition, they showed similar stability and electro-chemical properties.

View Article: PubMed Central - PubMed

Affiliation: Chemical Engineering Department, Universidad de los Andes, Bogotá, Colombia.

ABSTRACT
Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametespubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69 ± 0.28 U mg(-1) of protein for the crude extract, and 0.08 ± 0.001 and 2.86 ± 0.05 U mg(-1) of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct.

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Laccase activity profiles during T. pubescens CBS 696.94 culture under SmC (a) and SSC (b) on coffee husk.Culture media was enriched with different copper concentrations: 0 (Δ), 0.5 (□), 2 (○) and 5 (◊) mM, and laccase activity was determined by using syringaldazine as substrate. Filled symbols refer to the glucose concentration at the respective copper concentration. All the assays were performed in triplicate.
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pone-0073721-g001: Laccase activity profiles during T. pubescens CBS 696.94 culture under SmC (a) and SSC (b) on coffee husk.Culture media was enriched with different copper concentrations: 0 (Δ), 0.5 (□), 2 (○) and 5 (◊) mM, and laccase activity was determined by using syringaldazine as substrate. Filled symbols refer to the glucose concentration at the respective copper concentration. All the assays were performed in triplicate.

Mentions: Coffee husk, soybean pod husk, and cedar sawdust were used as substrate for the production of laccase by T. pubescens under SmC or SSC conditions, while syringaldazine was used to monitor laccase production during the fungus culture under the studied conditions. Hydrogen peroxide was not detected in any of the evaluated samples, which discard the interference of peroxidases in the syringaldazine assay. Laccase activity and glucose concentration time course for T. pubescens grown on coffee husk under SmC and SSC at different copper concentrations are presented in Figure 1. Although, a similar time course was observed for laccase activity and glucose concentration of soybean pod husk and cedar sawdust cultures, differences on the day of maximum lacasse activity as well as in the maximum activity value were observed among the agro-industrial byproducts (Table 2). In general, it was noticed that laccase activity was detectable around the seventh and fourth day of culture for SmC and SSC, respectively. After the eighth culture day, glucose was depleted from the culture medium and laccase activity increased sharply.


Production of Trametes pubescens laccase under submerged and semi-solid culture conditions on agro-industrial wastes.

Gonzalez JC, Medina SC, Rodriguez A, Osma JF, Alméciga-Díaz CJ, Sánchez OF - PLoS ONE (2013)

Laccase activity profiles during T. pubescens CBS 696.94 culture under SmC (a) and SSC (b) on coffee husk.Culture media was enriched with different copper concentrations: 0 (Δ), 0.5 (□), 2 (○) and 5 (◊) mM, and laccase activity was determined by using syringaldazine as substrate. Filled symbols refer to the glucose concentration at the respective copper concentration. All the assays were performed in triplicate.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760920&req=5

pone-0073721-g001: Laccase activity profiles during T. pubescens CBS 696.94 culture under SmC (a) and SSC (b) on coffee husk.Culture media was enriched with different copper concentrations: 0 (Δ), 0.5 (□), 2 (○) and 5 (◊) mM, and laccase activity was determined by using syringaldazine as substrate. Filled symbols refer to the glucose concentration at the respective copper concentration. All the assays were performed in triplicate.
Mentions: Coffee husk, soybean pod husk, and cedar sawdust were used as substrate for the production of laccase by T. pubescens under SmC or SSC conditions, while syringaldazine was used to monitor laccase production during the fungus culture under the studied conditions. Hydrogen peroxide was not detected in any of the evaluated samples, which discard the interference of peroxidases in the syringaldazine assay. Laccase activity and glucose concentration time course for T. pubescens grown on coffee husk under SmC and SSC at different copper concentrations are presented in Figure 1. Although, a similar time course was observed for laccase activity and glucose concentration of soybean pod husk and cedar sawdust cultures, differences on the day of maximum lacasse activity as well as in the maximum activity value were observed among the agro-industrial byproducts (Table 2). In general, it was noticed that laccase activity was detectable around the seventh and fourth day of culture for SmC and SSC, respectively. After the eighth culture day, glucose was depleted from the culture medium and laccase activity increased sharply.

Bottom Line: The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2).Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3.In addition, they showed similar stability and electro-chemical properties.

View Article: PubMed Central - PubMed

Affiliation: Chemical Engineering Department, Universidad de los Andes, Bogotá, Colombia.

ABSTRACT
Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametespubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69 ± 0.28 U mg(-1) of protein for the crude extract, and 0.08 ± 0.001 and 2.86 ± 0.05 U mg(-1) of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct.

Show MeSH
Related in: MedlinePlus