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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 stability profiles at optimal pH and temperature conditions for crude extracts (solid line), Lac1 (dashed line), and Lac2 (spotted line) obtained from semisolid culture of T. pubescens on coffee husk (◊) and cedar sawdust (•).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.
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pone-0073721-g006: Laccase stability profiles at optimal pH and temperature conditions for crude extracts (solid line), Lac1 (dashed line), and Lac2 (spotted line) obtained from semisolid culture of T. pubescens on coffee husk (◊) and cedar sawdust (•).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.

Mentions: It was observed that the activity loss depends on the substrate source used to grow T. pubescens. An activity loss of 55% was observed for crude extracts and purified fractions from coffee husk and soybean pod husk cultures, while an activity loss of 70% was observed for the crude extract and purified fractions from cedar sawdust culture. Nevertheless, Lac2 showed the lowest activity reduction, with values of 10, 40 and 55% for coffee husk, soybean pod husk, and cedar sawdust, respectively, while Lac1 presented an activity loss of 34, 50 and 60% for coffee husk, soybean pod husk, and cedar sawdust, respectively. Despite the differences in laccase activity among agro-industrial wastes and between crude extracts and purified fractions, all the profiles showed a sharp decrease during the first 8 h while a reduced activity loss was observed during the last 24 h of the assay (Figure 6).


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 stability profiles at optimal pH and temperature conditions for crude extracts (solid line), Lac1 (dashed line), and Lac2 (spotted line) obtained from semisolid culture of T. pubescens on coffee husk (◊) and cedar sawdust (•).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-g006: Laccase stability profiles at optimal pH and temperature conditions for crude extracts (solid line), Lac1 (dashed line), and Lac2 (spotted line) obtained from semisolid culture of T. pubescens on coffee husk (◊) and cedar sawdust (•).Laccase stability was evaluated by using ABTS. All the assays were performed in triplicate.
Mentions: It was observed that the activity loss depends on the substrate source used to grow T. pubescens. An activity loss of 55% was observed for crude extracts and purified fractions from coffee husk and soybean pod husk cultures, while an activity loss of 70% was observed for the crude extract and purified fractions from cedar sawdust culture. Nevertheless, Lac2 showed the lowest activity reduction, with values of 10, 40 and 55% for coffee husk, soybean pod husk, and cedar sawdust, respectively, while Lac1 presented an activity loss of 34, 50 and 60% for coffee husk, soybean pod husk, and cedar sawdust, respectively. Despite the differences in laccase activity among agro-industrial wastes and between crude extracts and purified fractions, all the profiles showed a sharp decrease during the first 8 h while a reduced activity loss was observed during the last 24 h of the assay (Figure 6).

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