Process optimization for enhanced production of cellulases form locally isolated fungal strain by submerged fermentation

Cellulase has myriad applications in various sectors like pharmaceuticals, textile, detergents, animal feed and bioethanol production, etc. The current study focuses on the isolation, screening and optimization of fungal strain through one factor at a time technique for enhanced cellulase production. In current study sixteen different fungal cultures were isolated and the culture which quantitatively exhibits higher titers of cellulase activity was identified both morphologically and molecularly by 18S rDNA and designated as Aspergillus niger ABT11. Different parameters like fermentation medium, volume, temperature, pH and nutritional components were optimized. The highest CMCase and FPase activities was achieved in 100ml of M5 medium in the presence of 1% lactose and sodium nitrate at 30 oC, pH5 after 72 hours. The result revealed A. niger can be a potential candidate for scale up studies.

Albizia lebbeck fruit pods as a substrate for the production of lignocellulolytic enzymes by Aspergillus niger

Aspergillus niger isolated from the soil was investigated for its capability to produce various lignocellulolyticenzymes, such as LiP, endoglucanase, FPase, xylanase by solid-state fermentation, using Albizia lebbeck fruitpods as a substrate. The chemical composition of the fruit pods was studied, and the production pattern of theenzymes was examined by growing the fungi for 25 days. The LiP activity was low, whereas a good productionof endoglucanase, FPase, and xylanase enzymes was noted. A dye decolorization capacity of the A. niger wasalso studied with Congo red. Therefore, A. lebbeck fruit pods which are considered as waste and burnt off canbe utilized for the production of holocellulolytic enzymes using A. niger.

Cellulolytic ability of a promising Irpex lacteus (Basidiomycota: Polyporales) strain from the subtropical rainforest of Misiones province, Argentina / Capacidad celulolítica de una cepa prometedora de Irpex lacteus (Basidiomycota: Polyporales) de la selva subtropical de la provincia de Misiones, Argentina

Abstract The cellulolytic activity of fungi growing in the subtropical rainforest of Misiones (Argentina) represents a challenge in the technological development of the production of cellulosic bioethanol in the region using native sources. These fungi are promising to obtain sustainable enzyme cocktails using their enzymes. Cellulolytic ability of 22 white-rot fungi isolated from the subtropical rainforest of Misiones-Argentina in agar medium with two types of cellulosic substrates, carboxy-methylcellulose or crystalline cellulose, were comparatively analyzed, and the activity of two cellulolytic enzymes was evaluated in liquid medium. Although all isolates were able to grow and degrade both substrates in agar medium, and to produce total cellulase Filter paper (FPase) and endo-β-1,4-glucanase (EG) activities in broth, the isolate Irpex sp. LBM 034 showed the greatest enzymatic levels (FPase, 65.45 U L-1; EG, 221.21 U L-1). Therefore, the ITS sequence of this fungus was sequenced and analyzed through a phylogenetic analysis. These results indicate that the isolate LBM 034, corresponding to Irpex lacteus, has a promising cellulolytic ability and enzymes such as EG useful in sustainable saccharification of cellulosic materials in the region. Rev. Biol. Trop. 66(3): 1034-1045. Epub 2018 September 01.

Resumen La actividad celulolítica de hongos autóctonos asociados a la selva subtropical de Misiones (Argentina) representa un desafío en el desarrollo tecnológico de la producción de bioetanol celulósico en la región, mediante el uso de recursos nativos. Los sistemas enzimáticos de estos hongos tienen potencial aplicación en la obtención de cocteles enzimáticos rentables. La habilidad celulolítica de 22 hongos causantes de pudrición blanca se analizó comparativamente, que fueron aislados de la selva subtropical de Misiones-Argentina, en cultivos agarizados con dos tipos de sustratos celulósicos, carboxi-metilcelulosa o celulosa cristalina. También se evaluó la actividad de dos enzimas celulolíticas en cultivos líquidos. Aunque todos los aislamientos fueron capaces de crecer y degradar ambos sustratos en medio agarizado y revelar actividad celulolítica total y endo-β-1,4-glucanasa en cultivo líquido, el aislamiento Irpex sp. LBM 034 mostró las mayores actividades en papel de filtro con 65.45 U L-1 y endo-β-1,4-glucanasa con 221.21 U L-1, respectivamente. Por tanto, se secuenció y analizó la secuencia ITS de este hongo a través de un análisis filogenético. Estos resultados indicaron que el aislamiento LBM 034, correspondiente a Irpex lacteus, tiene una habilidad celulolítica prometedora en la producción de enzimas con actividad endo-β-1,4-glucanasa, útil en la sacarificación sustentable de materiales celulósicos de la región.

Measurement of Cellulolytic Potential of Cellulase Producing Bacteria

The bioconversion of cellulose and hemicellulose to soluble sugars is important for global stabilization and for a sustainable human society. Here, hundreds of cellulolytic bacteria were found in soil, compost and animal waste slurry of our environment. Bacillus spp. are aerobic cellulolytic bacteria. Here, two Bacillus strains 2414, 2579 (T) and their mixed culture utilized for measuring the cellulolytic potential. The capability of cellulolytic potential was analyzed by enriching the basal salt media with Whatman no.1 filter paper as a substrate for cellulose degradation. Here, Cellulose-degrading potential of Bacillus strains was measured by measuring the diameter of a clear zone around the colony and its hydrolytic value on cellulose Congo-Red agar media. The extracellular cellulase activities ranged from 0.08233 to 0.44 IU/mL for FPase and 0.243 to 0.595 IU/mL for endoglucanase assay. The maximum activities range of β-glucosidase or cellobiase activity was 0.6 to1.5 1U/ml. The maximum xylanase activities value Bacillus cellulolysticus 2579 (T), Bacillus subtilis 2414 and their mixed culture were 12.0,11.5 and 12.5 unit/mL, respectively. All the enzymes were stable at an optimum pH range value of 3.0-7.0 and temperature range of 30˚C-50˚C. The maximum filter paper degradation percentage was estimated to be 71.76% by mixed culture after 48hrs of incubation period, it was observed that the maximum filter paper degradation was done by mixed culture than Bacillus strains. Biodiesel production was estimated by following the EN-14103 method and ester content was calculated on the basis of response factor with a minimum set value of ester content will be 96.5%.

Aspergillus sclerotiorum: riesgo para la herencia cultural y la salud / Aspergillus sclerotiorum: a risk to cultural heritage and health Abstract / Aspergillus sclerotiorum: risco para a herança cultural e para a saúde

A partir de una mancha de enmohecido de un libro del Archivo "Coronado" con valor patrimonial, se obtuvo una cepa fúngica mediante aislamiento en medio de cultivo micológico. La cepa fué Aspergillus sclerotiorum y se identificó por secuenciación de ADN ribosomal. Se evaluó y confirmó su capacidad deteriorante sobre el papel, al determinar cualitativamente las actividades enzimáticas celulolítica, proteolítica y amilolítica, así como la producción de pigmentos y ácidos. Se evaluaron cuantitativamente sus actividades enzimáticas celulasa total sobre papel de filtro (FPasa) y &beta-endoglucanasa, FPasa y &beta-endoglucanasa; confirmando actividades bajas. Aunque su capacidad deteriorante es discreta, resulta ser amenza para la conservación del documento y potencialmente peligroso para la salud de las personas que consultan y archivan.

By performing a mycological culture, we isolated a fungal strain from a mold patch in a book of great heritage value from the "Coronado" archives. Ribosomal DNA sequencing identified the strain as Aspergillus sclerotiorum. By qualitatively determining its cellulolytic, proteolytic and amylolytic and enzymatic activities, as well as the production of pigments and acids, we confirmed its paper deteriorating abilities. Quantitatively, we evaluated its total cellulase enzyme activities on filter paper (FPase) and &beta-endoglucanase, and FPase and &beta-endoglucanase; confirming low activities. Although its deteriorating abilities are weak, it poses a threat to the preservation of the document and is a potential health hazard to the people who refer to and archive these books.

A partir de uma mancha de bolor de um livro do Arquivo "Coronado" com valor patrimonial, obteve-se uma cepa fúngica mediante isolamento em meio de cultura micologica. A cepa foi Aspergillus sclerotiorum e identificou-se por sequenciação de ADN ribossomal. Avaliou-se e confirmou-se a sua capacidade deteriorantesobre o papel, ao determinar qualitativamente as atividades enzimáticas celulósica, proteolitica e amiolitica, assim como a produção de epigmentos e ácidos. Avaliaram-se quantitativamente as suas atividades enzimáticas celulase toral sobre o papel de filtro (FPasa) e &beta-endoglucanasa, FPasa e &beta-endoglucanasa; confirmando atividades baixas. Apesar da sua capacidade deteriorante ser discreta, resulta ser ameaçante para a conservação do documento e potencialmente perigoso para a saúde das pessoas que consultam e arquivam.

Saccharification of Sugarcane Bagasse by Enzymatic Treatment for bioethanol production

Aims: The escalating demands for traditional fossil fuels with unsecured deliverance and issues of climate change compel the researchers to develop alternative fuels like bioethanol. This study examines the prospect of biofuel production from high carbohydrate containing lignocellulosic material, e.g. sugarcane bagasse through biological means. Methodology and Results: Cellulolytic enzymes were collected from the culture filtrate of thermotolerant Trichoderma viride grown on variously pre-treated sugarcane bagasse. CMCase and FPase enzyme activities were determined as a measure of suitable substrate pre-treatment and optimum condition for cellulolytic enzyme production. The highest CMCase and FPase activity was found to be 1.217 U/ml and 0.109 U/ml respectively under the production conditions of 200 rpm, pH 4.0 and 50 °C using steamed NaOH treated bagasse as substrate. SEM was carried out to compare and confirm the activity of cellulolytic enzymes on sugarcane bagasse. Saccharification of pre-treated bagasse was carried out with crude enzymes together using a two-factor experimental design. Under optimized conditions the pre-treated bagasse was saccharified up to 42.7 % in 24 h. The hydrolysate was concentrated by heating to suitable concentration and then used for fermentation by an indigenous isolate of Saccharomyces cerevisiae. With 50 and 80 % brix containing liquor the concentration of alcohol was 0.579 % and 1.15 % respectively. Conclusion, significance and impact of study: This is the first report in Bangladesh for the production of cellulosic ethanol using local isolates. Though the rate of alcohol production was very low, a great impetus in this field can maximize the production thereby meet the demand for fuel in future.