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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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Effect of temperature (a) and pH (b) on laccase activity for the UF retentate obtained by semisolid culture of T. pubescens on coffee husk (□), soybean pod husk (◊), and cedar sawdust (Δ).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.
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pone-0073721-g004: Effect of temperature (a) and pH (b) on laccase activity for the UF retentate obtained by semisolid culture of T. pubescens on coffee husk (□), soybean pod husk (◊), and cedar sawdust (Δ).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.

Mentions: Temperature and pH effect on laccase activity was determined for the UF retentate, and Lac1 and Lac2 fractions obtained along the purification of the enzymatic crude extracts that resulted after the SSC of T. pubescens on the selected agro-industrial wastes. Laccase activity for the UF retentate under different temperature and pH conditions is presented in Figure 4 and Table 4. The maximum relative activity of the evaluated UF retentates was observed at 50-55 °C, and at a pH range from 2.0 to 4.0. However, UF retentate from cedar sawdust culture behaved different to the UF retentate from coffee husk and soybean pod husk cultures. In this sense, the UF retentate from cedar sawdust culture presented the lowest enzyme activity at temperatures ≤ 40 °C, but at 70 °C presented the highest enzyme activity, ~90% of its initial value, in comparison to the other two UF retentates (Figure 4).


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)

Effect of temperature (a) and pH (b) on laccase activity for the UF retentate obtained by semisolid culture of T. pubescens on coffee husk (□), soybean pod husk (◊), and cedar sawdust (Δ).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0073721-g004: Effect of temperature (a) and pH (b) on laccase activity for the UF retentate obtained by semisolid culture of T. pubescens on coffee husk (□), soybean pod husk (◊), and cedar sawdust (Δ).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.
Mentions: Temperature and pH effect on laccase activity was determined for the UF retentate, and Lac1 and Lac2 fractions obtained along the purification of the enzymatic crude extracts that resulted after the SSC of T. pubescens on the selected agro-industrial wastes. Laccase activity for the UF retentate under different temperature and pH conditions is presented in Figure 4 and Table 4. The maximum relative activity of the evaluated UF retentates was observed at 50-55 °C, and at a pH range from 2.0 to 4.0. However, UF retentate from cedar sawdust culture behaved different to the UF retentate from coffee husk and soybean pod husk cultures. In this sense, the UF retentate from cedar sawdust culture presented the lowest enzyme activity at temperatures ≤ 40 °C, but at 70 °C presented the highest enzyme activity, ~90% of its initial value, in comparison to the other two UF retentates (Figure 4).

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