Evaluación del consorcio entre Pleurotus ostreatus, Trametes versicolor y bacterias aeróbicas para remoción de colorantes sintéticos / Evaluation of the consortium between Pleurotus ostreatus, Trametes versicolor and aerobic bacteria for removal of synthetic dye

RESUMEN Los residuos líquidos producidos al elaborar tinciones biológicas contienen mezclas de compuestos químicos y microorganismos, que generan un elevado impacto ambiental si no son tratados adecuadamente. Por esta razón, en el presente trabajo se evaluaron a Pleurotus ostreatus, Trametes versicolor, Enterobacter xianfangensis, Pseudomonas azotoformans, Pseudomonas sp., Bacillus subtilis y Pseudomonas fluorescens, para el tratamiento de un residuo líquido que contenía colorantes trifenilmetánicos y azóicos, a escala de laboratorio. Inicialmente, se seleccionaron las cepas con menor efecto antagónico y se determinó su potencial para producir las enzimas Lacasa, Manganeso Peroxidasa y Lignino Peroxidasa, al emplear sustratos inductores y mezclas de colorantes. Para el consorcio fúngico/bacteriano la disminución de las unidades de color y demanda química de oxígeno fueron del 99 % y 70 % a las 96 h. La remoción de estos parámetros se relacionó con la interacción positiva e incremento de las poblaciones de hongos, bacterias y la producción de enzimas ligninolíticas, obteniendo valores a las 96 h de 7.0 y 14.0 unidades logarítmicas para hongos y bacterias, con unas actividades enzimáticas de 75 U/L, 205 U/L y 0.63 U/L para Lacasa, MnP y LiP, respectivamente. Con el presente trabajo se demostró que con el uso consorcios fúngicos/bacterianos se incrementa la remoción de colorantes y se disminuye el tiempo de proceso. Sugiriendo que estos microorganismos podrían ser evaluados en plantas de tratamiento que integren diferentes unidades de tratamiento para optimizar la remoción de contaminantes con baja biodegradabilidad.

ABSTRACT The liquid wastes generated when biological stains are prepared, contain a mixture of chemical compounds and microorganisms, with high environmental impact. For this reason, Pleurotus ostreatus, Trametes versicolor, Enterobacter xianfangensis, Pseudomonas azotoformans, Pseudomonas sp., Bacillus subtilis and Pseudomonas fluorescens, were used to evaluate the treatment of a liquid waste containing triphenylmethane and azo dyes, on a laboratory scale. Initially, the strains with less antagonistic effect among them were selected for their potential to produce enzymes as Laccase, Manganese Peroxidase and Lignin Peroxidase. The enzymatic activity was determined by using inducing substrates and dye mixtures. For fungal/bacterial consortium, the decrease in color, Chemical Oxygen Demand and in Biochemical Oxygen demand was of 99 %, 70 % and 65 % at 96 h, respectively. The removal of these parameters was related to the positive interaction between the populations of fungi, bacteria and the production of ligninolytic enzymes, obtaining values of 7.0 and 14.0 logarithmic units for fungi and total bacteria at 96 h with enzymatic activities of 75 U/ L, 205 U/L and 0.63 U/L for Laccase, MnP and LiP. The present work demonstrates that using of fungal/bacterial consortia, the removal of dyes is increased, and the process time is decreased. Suggesting that these microorganisms could be evaluated in treatment plants that integrate different treatment units to optimize the removal of contaminants with low biodegradability.

A novel textile wastewater treatment using ligninolytic co-culture and photocatalysis with TiO2 / Un novedoso tratamiento de aguas residuales de la industria textil usando co-cultivos ligninolíticos y fotocatálisis con TiO2 / Um novo tratamento para águas residuais têxtis usando co-cultura ligninolítica e fotocatálise com TiO2

Abstract Textile industries produce effluent wastewater that, if discharged, exerts a negative impact on the environment. Thus, it is necessary to design and implement novel wastewater treatment solutions. A sequential treatment consisting of ligninolytic co-culture with the fungi Pleurotus ostreatus and Phanerochaete crhysosporium (secondary treatment) coupled to TiO2 /UV photocatalysis (tertiary treatment) was evaluated in the laboratory in order to discolor, detoxify, and reuse textile effluent wastewater in subsequent textile dyeing cycles. After 48 h of secondary treatment, up to 80% of the color in the wastewater was removed and its chemical and biochemical oxygen demands (COD, and BOD5) were abated in 92% and 76%, respectively. Laccase and MnP activities were central to color removal and COD and BOD5 abatement, exhibiting activity values of 410 U L-1 and 1 428 U L-1, respectively. Subjecting wastewater samples to 12 h of tertiary treatment led to an 86% color removal and 73% and 86% COD and BOD5 abatement, respectively. The application of a sequential treatment for 18 h improved the effectiveness of the wastewater treatment, resulting in 89% of color removal, along with 81% and 89% COD and BOD5 abatement, respectively. With this sequential treatment a bacterial inactivation of 55% was observed. TiO2 films were reused continuously during two consecutive treatment cycles without thermic reactivation. Removal percentages greater than 50% were attained. Acute toxicity tests performed with untreated wastewater led to a lethality level of 100% at 50% in Hydra attenuata and to a growth inhibition of 54% at 50% in Lactuca sativa. Whereas sequentially treated wastewater excreted a 13% lethality at 6.25% and an inhibition of 12% at 75% for H. attenuata and L. sativa, respectively. Finally, sequentially treated wastewater was reused on dyeing experiments in which 0.86 mg g-1 adsorbed dye per g of fabric, that is equivalent to 80% of dye adsorption.

Resumen Las industrias textiles producen un efluente que, si es descargado, ejerce un impacto negativo en el ambiente. Así, es necesario diseñar e implementar soluciones novedosas para el tratamiento de estos desechos. Con el fin de decolorar, detoxificar y reutilizar aguas residuales producidas en la industria textil en ciclos de tinturado subsecuentes, se evaluó en laboratorio un tratamiento secuencial, consistente en un cultivo ligninolítico con los hongos Pleurotus ostreatus y Phanerochaete crhysosporium (tratamiento secundario) acoplado con fotocatálisis TiO2/UV (tratamiento terciario). Después de 48 horas de tratamiento secundario, se removió hasta un 80% del color del agua residual y las Demandas Química y Bioquímica de Oxígeno (DQO y DBO5) fueron reducidas en 92 y 76% respectivamente. Las actividades Lacasa y MnP (410 U L-1 y 1428 U L-1, respectivamente) fueron centrales en la remoción de color y en la reducción de DQO y DBO5. Someter muestras de agua residual a 12 horas de tratamiento terciario resultó en una remoción de 86% de color, 73% de DQO y 86% de DBO. La aplicación de un tratamiento secuencial por 18 h incrementó su efectividad, lo cual resultó en 89% de remoción de color, además de 81% y 89% de remoción de DQO y DBO5 respectivamente. Con este tratamiento secuencial se observó una inactivación bacteriana del 55%. Las películas de TiO2 se reutilizaron continuamente durante 2 ciclos de tratamiento consecutivos sin reactivación térmica. Se alcanzaron porcentajes de remoción mayores al 50%. Las pruebas de toxicidad aguda llevadas a cabo con agua residual cruda produjeron un nivel de letalidad del 100% a 50% en Hydra attenuata y una inhibición de crecimiento del 54% a 50% en Lactuca sativa. El agua residual tratada secuencialmente produjo una letalidad del 13% a 6.25% y una inhibición del 12% a 75% para H. attenuata y L. sativa, respectivamente. Finalmente, el agua residual tratada secuencialmente fue reutilizada en experimentos de tinturado en los que la cantidad de colorante absorbido fue de 0.86 mg g-1 por g de tela, lo cual es equivalente a 80% de adsorción.

Resumo A indústria têxtil produz águas residuais que causam um impacto negativo ao meio ambiente quando são descartadas. Portanto, é necessário projetar e implementar novas soluções para seu tratamento. Um tratamento sequencial que consiste na co-cultura ligninolítica com os fungos Pleurotus ostreatus e Phanerochaete crhysosporium (tratamento secundário) acoplada à fotocatálise com TiO2/UV foi avaliado no laboratório para descolorir, desintoxicar e reutilizar água residual têxtil em ciclos subsequentes de tintura. Após 48 h de tratamento secundário, aproximadamente 80% da cor foi removida da água residual e sua demanda química e bioquímica de oxigênio (COD, e BOD5) foram diminuídas em 92% e 76%, respectivamente. A atividade de Laccase e MnP foram centrais na remoção da cor e diminuição de COD e BOD5, mostrando valores de atividade de 410 U L-1 e 1428 U L-1, respectivamente. Submeter as amostras de água residual a 12 h de tratamento terciário levou a uma remoção de 86% da cor e uma diminuição de 73% e 86% de COD e BOD5 respectivamente. A aplicação do tratamento sequencial por 18 h melhorou a efetividade do tratamento de águas residuais, resultando em 89% de remoção de cor e uma diminuição de 81% e 89% de COD e BOD5, respectivamente. Com este tratamento sequencial, observamos uma inativação bacteriana de 55%. As películas de TiO2 foram reutilizadas continuamente durante dois tratamentos consecutivos sem reativação térmica. Atingimos porcentagens de remoção acima de 50%. Os testes de toxicidade aguda feitos em água residual sem tratar causaram níveis de letalidades de 100% a 50% em Hydra attenuata e uma inibição do crescimento de 54% a 50% em Lactuca sativa. Enquanto água residual tratada sequencialmente causou uma letalidade de 13% a 6.25% e uma inibição de 12% a 75% para H. attenuata e L. sativa, respectivamente. Finalmente, a água residual tratada sequencialmente foi reutilizada em experimentos de tintura nos quais absorveu 0.86 mg g-1 por g de tecido, o que é equivalente a uma absorção de 80%.

Production of lignocellulolytic enzymes from floriculture residues using Pleurotos ostreatus / Producción de enzimas lignocelulolíticas a partir de residuos de floricultura empleando Pleurotus ostreatus / Produçâo de enzimas lignoceluliticas a partir de residuos de floricultura utilizando Pleurotus ostreatus

Floriculture is a vital agro-industrial sector in the Colombian economy; the export of flowers positively impacts employment and the balance of trade. However, this industry could negatively impact the environment if its waste products are not handled properly. These flower residues, rich in lignin, hemicellulose and cellulose, could be a cost-effective raw material to produce enzymes. Here, we evaluate the production of lignocellulolytic enzymes by degradation of Chrysanthemum and Rosa residues using Pleurotus ostreatus, and manganese sulfate and copper sulfate as inductors. From the two residues, we obtained laccase, manganese peroxidase, endoglucanase, exoglucanase, and (3-glucosidase. The use of inductors, favored all enzyme activities except for (3-glucosidase. The enzymes that displayed the highest activity were laccase (4,693.4 U/L and 2,640 U/L from the residues of Chrysanthemum and Rosa, respectively) and (3-glucosidase (9,513 U/L and 6,811.9 U/L). The enzyme that showed the lowest activity was endoglucanase (11.5 U/L and 15.4 U/L). Under the conditions evaluated, the best substrate for enzyme production is Chrysanthemum wastes; the extracts obtained had higher enzymatic activity than the extracts from Rosa residues.

En Colombia la floricultura es un sector agro-industrial importante, con impactos positivos en el empleo y la balanza comercial. Sin embargo, tiene impacto negativo en el medio ambiente porque genera alto volumen de residuos. Estos residuos, ricos en lignina, hemicelulosa y celulosa, podrían ser una materia prima de bajo costo para la producción de enzimas. En este trabajo se estudió en la producción de enzimas lignocelulíticas por la degradación con Pleurotus ostreatus de residuos de Chrysanthemum y Rosa, usando como inductores sulfato de manganeso y de cobre. A partir de ambos residuos se obtuvieron lacasa, manganeso peroxidasa, endoglucanasa, exoglucanasa y p-glucosidasa. Los inductores favorecieron todas las actividades enzimáticas, excepto para p-glucosidasa. Las enzimas que tuvieron mayores actividades fueron lacasa (4,693.4 U/L y 2,640 U/L a partir del residuo de Chrysanthemum y Rosa, respectivamente) y p-glucosidasa (9,513 U/L y 6,811.9 U/L). La enzima que tuvo menor actividad fue endoglucanasa (11.5 U/L y 15.4 U/L). Bajo las condiciones evaluadas, el mejor residuo para producción de enzimas fue Chrysanthemum, porque los extractos tuvieron mayor actividad enzimática que los producidos a partir de Rosa.

Na Colombia, a floricultura è um importante setor da indùstria agrícola, com impactos positivos sobre o sector laboral na balança comercial. Além disto, tem impacto negativo sobre o meio ambiente, pois gera grandes volumes de resíduos. Estes resíduos, com altos conteùdos em lignina, hemicelulose e celulose, podem ser uma matèria-prima de baixo custo para a produçâo de enzimas. Neste traballio foi estudada a produçâo da enzimas lignoceluliticas pela degradaçâo com Pleurotus ostreatus de residuos de Chrysanthemum e Rosa, utilizando como indutores sulfatos de cobre e manganês. A partir destes resíduos foram obtidos lacase, manganês peroxidase, endoglucanase, exoglucanase e β-glicosidase. O uso de indutores favoreceu as atividades enzimáticas, exceto β-glicosidase. As enzimas que apresentaram atividades mais elevadas foram lacase (4,693.4 U/L e 2,640 U/L a partir do Chrysanthemum e Rosa, respetivamente) e β-glicosidase (9,513 U/L e 6,81.9 U/L). A enzima que apresentou menor atividade foi a endoglucanase (11.5 U/L e 15.4 U/L). Nas condiçôes testadas, o melhor resíduo para a produçâo da enzima foi Chrysanthemum, porque os extratos tinham uma atividade enzimática mais elevada que aqueles produzidos a partir de Rosa.

Computational analysis and low-scale constitutive expression of laccases synthetic genes GlLCC1 from Ganoderma lucidum and POXA 1B from Pleurotus ostreatus in Pichia pastoris.

Lacasses are multicopper oxidases that can catalyze aromatic and non-aromatic compounds concomitantly with reduction of molecular oxygen to water. Fungal laccases have generated a growing interest due to their biotechnological potential applications, such as lignocellulosic material delignification, biopulping and biobleaching, wastewater treatment, and transformation of toxic organic pollutants. In this work we selected fungal genes encoding for laccase enzymes GlLCC1 in Ganoderma lucidum and POXA 1B in Pleurotus ostreatus. These genes were optimized for codon use, GC content, and regions generating secondary structures. Laccase proposed computational models, and their interaction with ABTS [2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] substrate was evaluated by molecular docking. Synthetic genes were cloned under the control of Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase (GAP) constitutive promoter. P. pastoris X-33 was transformed with pGAPZαA-LaccGluc-Stop and pGAPZαA-LaccPost-Stop constructs. Optimization reduced GC content by 47 and 49% for LaccGluc-Stop and LaccPost-Stop genes, respectively. A codon adaptation index of 0.84 was obtained for both genes. 3D structure analysis using SuperPose revealed LaccGluc-Stop is similar to the laccase crystallographic structure 1GYC of Trametes versicolor. Interaction analysis of the 3D models validated through ABTS, demonstrated higher substrate affinity for LaccPost-Stop, in agreement with our experimental results with enzymatic activities of 451.08 ± 6.46 UL-1 compared to activities of 0.13 ± 0.028 UL-1 for LaccGluc-Stop. This study demonstrated that G. lucidum GlLCC1 and P. ostreatus POXA 1B gene optimization resulted in constitutive gene expression under GAP promoter and α-factor leader in P. pastoris. These are important findings in light of recombinant enzyme expression system utility for environmentally friendly designed expression systems, because of the wide range of substrates that laccases can transform. This contributes to a great gamut of products in diverse settings: industry, clinical and chemical use, and environmental applications.

Ethanol production by Saccharomyces cerevisiae using lignocellulosic hydrolysate from Chrysanthemum waste degradation.

Ethanol production derived from Saccharomyces cerevisiae fermentation of a hydrolysate from floriculture waste degradation was studied. The hydrolysate was produced from Chrysanthemum (Dendranthema grandiflora) waste degradation by Pleurotus ostreatus and characterized to determine the presence of compounds that may inhibit fermentation. The products of hydrolysis confirmed by HPLC were cellobiose, glucose, xylose and mannose. The hydrolysate was fermented by S. cerevisiae, and concentrations of biomass, ethanol, and glucose were determined as a function of time. Results were compared to YGC modified medium (yeast extract, glucose and chloramphenicol) fermentation. Ethanol yield was 0.45 g g(-1), 88 % of the maximal theoretical value. Crysanthemum waste hydrolysate was suitable for ethanol production, containing glucose and mannose with adequate nutrients for S. cerevisiae fermentation and low fermentation inhibitor levels.

Desarrollo de un bioadsorbente laminar con Phanerochaete chrysosporium hipertolerante al cadmio, al níquel y al plomo para el tratamiento de aguas / Development of a laminar bioadsorbent with hypertolerant Phanerochaete chrysosporium to Cd, Ni and Pb for wastewater treatment

Antecedentes. El uso de basidiomicetes para la remoción de metales es una alternativa con respecto a los métodos tradicionales, básicamente porque la biomasa actúa como un intercambiador natural al remover metales en solución. Objetivo. Desarrollar un bioadsorbente laminar empleando el hongo basidiomicete con mayor resistencia al cadmio (Cd), al níquel (Ni) y al plomo (Pb). Métodos. Se valoró la tolerancia de Trametes versicolor, Pleurotus ostreatus y Phanerochaete chrysosporium frente a concentraciones ascendentes de sulfato de sulfato de cadmio, acetato de plomo y cloruro de níquel. Se desarrolló un bioadsorbente a base de láminas de polietileno con biomasa fúngica, que se evaluó en columnas de burbujeo empleando agua residual sintética con los 3 metales con una concentración final de 300mg/l. Finalmente, con un experimento complementario en matraces Erlenmeyer, se valoró el efecto de mayor cantidad de biomasa sobre la eficiencia de remoción. Resultados. El hongo P. chrysosporium fue la cepa más tolerante a C4H6O4Pb (10.000mg/l), a Cl2Ni (300mg/l) y CdSO4·8H2O (1.500mg/l). En el reactor y bajo condiciones no ligninolíticas, el hongo removió el 69% de la demanda química de oxígeno, produjo enzimas como lignino peroxidasa (0,01U/l) y manganeso peroxidasa (0,6U/l) y se observó la acumulación de los metales en la pared. Al incrementar la biomasa a 1,6 (p/v) se favoreció la bioadsorción de los metales en la mezcla (el 57% para el Pb, el 74% para el Cd y el 98% para el Ni) y por separado (el 95% para el Pb, el 60% para el Cd y el 56% para el Ni). Se observó una competencia entre el Ni y el Pb por los ligandos de la pared. Conclusión. Se desarrolló un novedoso sistema laminar a base de biomasa viable de P. chrysosporium que posee gran área superficial y tolera elevadas concentraciones de Cd, Ni y Pb, como alternativa para la remoción de metales en aguas(AU)

Background. The use of basidiomycetes for metal removal is an alternative to traditional methods. In this, the biomass acts as a natural ionic exchanger removing metals from solution. Objective. To develop a laminar biosorbent using a basidiomycete fungus resistant to high Cd, Ni and Pb concentrations. Methods. The tolerance of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium was evaluated using increasing concentrations of the heavy metal salts, cadmium sulphate, lead acetate and nickel chloride. A biosorbent system was developed based on polyethylene sheets with a fungal biomass. It was evaluated in bubble columns using synthetic wastewater with the 3 metal salts at a rate of 300mg/l. Finally, in a complementary experiment using shake flasks, the effect of a higher amount of biomass related to the metal removal efficiency was evaluated. Results. P. chrysosporium strain was more tolerant to C4H6O4Pb (10,000mg/l), Cl2Ni (300mg/l) and CdSO4·8H2O (1,500mg/l). In a reactor, under non-ligninolytic conditions, the fungus removed 69% of the chemical oxygen demand and produced enzymes such as LiP (0.01U/l) and MnP (0.6U/l.). An accumulation of metals in the wall was observed. By increasing the biomass to 1.6 (w/v), the metal biosorption was favored in the mixture (57% Pb, 74% Cd, and 98% Ni) and separately (95% Pb, 60% Cd, and 56% Ni). Competition between Ni and Pb by ligands of the wall was observed. Conclusion. A novel laminar system based on P. chrysosporium viable biomass was developed. It has a large surface area and tolerance to high concentrations of Cd, Ni and Pb. It seems to be an alternative for the removal of metals from water(AU)

Fabrication and optical characterization of a high-quality fcc-opal-based photonic crystal grown by the vertical convective self-assembly method / Fabricación y caracterización óptica de cristales coloidales de alta calidad formados por esferas de SiO2 de 225 nm de diámetro con empaquetamiento fcc crecidos por el método convectivo de auto-ensamblado de deposición vertical / Fabricação e caracterização óptica de cristais coloidais de alta qualidade formados por esferas de SiO2 de 225 nm de diâmetro com embalagem fcc crescidos pelo método convectivo de automontagem de deposição vertical

Objective: Fabrication and optical characterization of close-packed 225 nm SiO2 -based colloidal crystals. Materials and methods: The vertical convective self-assembly method is used to grow high-quality 225 nm close-packed SiO2-based colloidal crystals. An annealing process (550°C) is made in order to improve the mechanical stability of the sample. Optical characterization is done by angle-resolved transmission spectroscopy (A-RTS) and structural characterization by Scanning Electron Microscopy (SEM). Results: Both, A-RTS and SEM, show that with the vertical convective self-assembly method, with the appropriate parameters of temperature of evaporation (60°C), volume fraction of the colloidal suspension (0.2% w/w) and acidity (pH=6), highly ordered close packed face centered cubic (fcc) SiO2 based colloidal crystals are obtained. Conclusions: The growth of high-quality (long range order and defect-free) face centered cubic opalbased photonic crystals is reported.

Objetivo: Fabricación y caracterización óptica de cristales coloidales de esferas de SiO2 de 225 nm de diámetro y empaquetamiento fcc. Materiales y métodos: Se producen cristales coloidales de 225 nm de diámetro de alta calidad mediante la técnica convectiva de auto-ensamblado de deposición vertical. Se hace, adicionalmente, un proceso de recocido (550°C) con el fin de mejorar la estabilidad mecánica de la estructura. La caracterización óptica se lleva a cabo por espectroscopia de transmisión resuelta en ángulo (A-RTS) y la caracterización estructural mediante microscopía electrónica de barrido (SEM). Resultados: Ambos, A-RTS y MEB, muestran que con el método utilizado y con los parámetros adecuados de temperatura de evaporación (60°C), fracción en volumen de la suspensión coloidal (0,2% w / w) y acidez (pH = 6) se obtienen cristales coloidales de SiO2 con empaquetamiento compacto fcc altamente ordenados. Conclusiones: Se reporta el crecimiento de ópalos artificiales con empaquetamiento compacto fcc altamente ordenados (libre de defectos y con ordenamiento a largo alcance).

Objetivo: Fabricação e caracterização óptica de cristais coloidais de esferas de SiO2 de 225 nm de diâmetro e embalagem fcc. Materiais e métodos: Foram produzidos cristais coloidais de 225 nm de diâmetro de alta qualidade através da técnica convectiva de automontagem de deposição vertical. Faz-se, adicionalmente, um processo de recozimento (550°C), a fim de melhorar a estabilidade mecânica da estrutura. A caracterização óptica foi realizada por espectroscopia de transmissão em ângulo (A-RTS) e a caracterização estrutural utilizando microscopia eletrônica de varredura (MEV). Resultados: Ambos, A-RTS e MEV mostram que com o método utilizado e com os parâmetros adequados de temperatura de evaporação (60°C), fração de volume da suspensão coloidal (0,2% w / w) e acidez (pH = 6) são obtidos cristais coloidais de SiO2 com embalagem compacto fcc altamente ordenados. Conclusões: Registrou-se o crescimento de opalas artificiais com embalagem compacto fcc altamente ordenadas (livres de defeitos e de ordem de longo alcance).

[Development of a laminar bioadsorbent with hypertolerant Phanerochaete chrysosporium to Cd, Ni and Pb for wastewater treatment]. / Desarrollo de un bioadsorbente laminar con Phanerochaete chrysosporium hipertolerante al cadmio, al níquel y al plomo para el tratamiento de aguas.

BACKGROUND: The use of basidiomycetes for metal removal is an alternative to traditional methods. In this, the biomass acts as a natural ionic exchanger removing metals from solution. OBJECTIVE: To develop a laminar biosorbent using a basidiomycete fungus resistant to high Cd, Ni and Pb concentrations. METHODS: The tolerance of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium was evaluated using increasing concentrations of the heavy metal salts, cadmium sulphate, lead acetate and nickel chloride. A biosorbent system was developed based on polyethylene sheets with a fungal biomass. It was evaluated in bubble columns using synthetic wastewater with the 3 metal salts at a rate of 300 mg/l. Finally, in a complementary experiment using shake flasks, the effect of a higher amount of biomass related to the metal removal efficiency was evaluated. RESULTS: P. chrysosporium strain was more tolerant to C4H6O4Pb (10,000mg/l), Cl2Ni (300mg/l) and CdSO4·8H2O (1,500 mg/l). In a reactor, under non-ligninolytic conditions, the fungus removed 69% of the chemical oxygen demand and produced enzymes such as LiP (0.01 U/l) and MnP (0.6 U/l.). An accumulation of metals in the wall was observed. By increasing the biomass to 1.6 (w/v), the metal biosorption was favored in the mixture (57% Pb, 74% Cd, and 98% Ni) and separately (95% Pb, 60% Cd, and 56% Ni). Competition between Ni and Pb by ligands of the wall was observed. CONCLUSION: A novel laminar system based on P. chrysosporium viable biomass was developed. It has a large surface area and tolerance to high concentrations of Cd, Ni and Pb. It seems to be an alternative for the removal of metals from water.

Inmovilización de hongos ligninolíticos para la remoción del colorante negro reactivo 5 / Immobilising lignilolytic fungus for removing reactive black 5 dye

La presencia de colorantes azoicos en aguas residuales de la industria textil es un problema ambiental y sanitario, porque muchos de estos compuestos son cancerígenos. Los tratamientos biológicos son una alternativa para la remoción de ese tipo de colorantes. En el presente trabajo se evaluó el efecto de tres hongos de podredumbre blanca, Trametes versicolor, Pleurotus ostreatus y Phanerochaete chrysosporium sobre la decoloración de un agua que contiene colorante negro reactivo 5 (NR5), ampliamente usado en la industria textil. Se estudió la inmovilización de estos hongos en dos soportes, espuma de poliuretano y estropajo (L. cylíndrica) para seleccionar el mejor soporte y el hongo con mayor capacidad para la decoloración. Ambos soportes fueron igualmente efectivos, pero se seleccionó estropajo por ser un producto natural. El hongo que generó los mayores porcentajes de decoloración en 4 días fue Trametes versicolor, con 96%, 98% y 98% para agua con concentración de NR5 300 ppm, 150 ppm y 75 ppm, respectivamente. La actividad lacasa para cada concentración de NR5 fue 8 U L-1, 7 U L-1 y 5 U L-1.

Waste water from the textile industry represents a major environmental and health problem because it contains azo dyes whose carcirogenic effect has been tested in research. Biological treatment represents a valuable alternative for removing these dyes. The effect of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium rot fungi on decoloration of water containing reactive black five (NR5) textile dye was evaluated in this work. Immobilising the fungi on polyurethane foam and luffa sponge (Luffa cylindrica) supports was studied in order to select the best support and the fungi having the best decolorisation. Both supports were equally effective; however, the luffa sponge was selected as being a natural product. Trametes versicolor produced the highest decolorisation percentages in four days (96%, 98% and 98% for 300 ppm, 150 ppm and 75 ppm NR5 concentrations, respectively) while lacase enzyme activity was 8 UL-1, 7 UL-1 and 5 UL-1 for each of them.

Inmovilización de bacillus licheniformis y Saccharomyces cerevisiae para la producción de etanol a partir de almidón de papa / Inmobilization of bacillus licheniformis and Saccharomyces cerevisiae for ethanol production from potato starch

Se evaluó un sistema discontinuo secuencial compuesto por células de Bacillus licheniformis y Saccharomyces cerevisiae para producción de etanol, utilizando en la segunda fase del proceso, un hidrolizado de almidón de papa, obtenido con el uso de células de B. licheniformis. Ambos microorganismos fueron inmovilizados en matriz de alginato de calcio al 3,2 por ciento y 2,5 por ciento (p/v), observando que a estas concentraciones se retiene la mayor cantidad de células (26x106 y 10x107 UFC/g) y permite la difusión de los productos, obteniendo 3,3 g/L de azúcares reductores y 642 UA/L (unidades amilolíticas) para B. licheniformis y 0,866 por ciento (v/v) de etanol con S. cerevisiae. Mediante un diseño factorial 22 se seleccionaron las condiciones de operación a escala de reactor para la producción del hidrolizado, encontrando que al cultivar a B. licheniformis con 3 v.v.m. y 150 r.p.m. se produjeron 3,7 g/L de azúcares reductores y 669 UA/L a las 4 horas de proceso. El hidrolizado se caracterizó por cromatografía HPLC determinando que es rico en oligómeros, dextrinas y que tiene baja concentración de glucosa y maltosa. El uso del hidrolizado para la producción de etanol, generó porcentajes bajos (0,47por ciento y 0,74 por ciento v/v), tanto en células libres como inmovilizadas, respectivamente.

We evaluated a sequential discontinuous system composed by Bacillus licheniformis and Saccharomyces cerevisiae forethanol production. For the second phase of the process potato starch hydrolyzed were used, which was obtained from B. licheniformis cells. Both microorganisms were immobilized in a calcium alginate matrix of 3,2% and 2,5% (w/v), where was observed that these concentrations retained the majority of the cells (26x106 and 10x107 UFC/g) and alloweddissemination of its products, gaining 3.3 g/L of reducing sugars and 642 AU/L (units Amylolytic) for B. licheniformis and 0,866% (v/v) ethanol with S. cerevisiae. By means of a 22 factorial design were selected operating conditions at a reactor scale for production of hydrolyzed, finding that by cultivating B. licheniformis with 3 v.v.m. and 150 r.p.m. there were 3.7 g/L of reducing sugars and 669 AU/L after 4 hours of the process. The hydrolyzed was characterized using HPLC chromatography, which determined that it is rich in oligomers and dextrin, and it has low concentration of glucose andmaltose. The use of hydrolyzed for ethanol production, generated low percentages (0,47% and 0,74% v/v) in free and immobilized cells respectively.

El pulque: características microbiológicas y contenido alcohólico mediante espectroscopia Raman / Pulque: Microbiological characteristics and alcohol content by Raman spectroscopy

A partir de cuatro muestras de pulque (bebida mexicana prehispánica) en diferentes etapas de fermentación denominadas aguamiel, semilla, contrapunta y corrida; se aislaron e identificaron un hongo levaduriforme, un cocobacilo Gram negativo y un bacilo Gram positivo pertenecientes a los géneros Saccharomyces sp, Zymomonas sp, y Lactobacillus sp. El consorcio desarrollado fue el responsable de la fermentación, alcohólica, ácida y viscosa propias de esta bebida tradicional Mexicana. El contenido alcohólico varió en función del tiempo de fermentación alcanzando porcentajes de 10.35 (v/v) y 9.01 (v/v) determinados por cromatografía de gases y espectroscopia Raman para la etapa denominada ®corrida¼. El producto terminado tiene un elevado contenido de proteínas (1 g/L), azúcares reductores (4.75 g/L) y poblaciones microbianas en ordenes de 45x108 UFC/mL para Saccharomyces sp, 41x107 UFC/mL para Zymomonas sp y 34x106 UFC/mL para Lactobacillus sp, que le dan a la bebida un elevado valor nutricional y los microorganismos aislados podrían tener un efecto benéfico sobre sistema digestivo al ser consumido por vía oral.