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DEF from Japanese cedar wood meal with Tween 80 and lipase. (A) DEF with 5 U/g-dry substrate lipase and (B) DEF with 1% (v/v) Tween 80 and 5 U/g-dry substrate lipase. Closed circles denote ethanol concentrations and open circles indicate glucose concentrations. All DEFs were carried out at 10 °C. All experiments were carried out in triplicate and error bars denote standard deviation values.
Source publication
The Antarctic basidiomycetous yeast Mrakia blollopis SK-4 can quite uniquely ferment various sugars under low temperature conditions. When strain SK-4 fermented lignocellulosic biomass using the direct ethanol fermentation (DEF) technique, approximately 30% to 65% of the theoretical ethanol yield was obtained without and with the addition of the no...
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Background:
Lignocellulosic biomass from plant biomass, especially softwoods, are well-known to present difficulties during attempts at hydrolysis due to their rigid structure. Pretreatment of lignocellulosic biomass with ionic liquids (ILs) is attractive as this requires to a low input of energy. However, IL pretreatment has the disadvantage of t...
Citations
... The highest ethanol produced in SSSF experiments was also with 15% (w/v) solid loading and adding 1.0% (w/v) Tween 80, which reached 26.24 ± 0.53 g/L, a productivity of 0.55 g/L.h, and an ethanol yield of 56.39%. Despite several studies showing that ethanol production increase when using Tween 80 [24,[51][52][53], this behavior was not found in this study for SSF and SSSF strategies. One hypothesis may be that even with the increase in glucose production by enzymatic hydrolysis, the surfactant decreased cell viability, impairing yeast growth and decreasing ethanol production. ...
Using lignocellulosic materials as a feedstock to produce ethanol is an alternative to cleaner energy production. The pretreatment and enzymatic hydrolysis steps play a role in breaking down the lignocellulosic structure and making the sugars available for fermentation. Non-ionic surfactants have been included in the cellulosic ethanol production process to increase ethanol titers. Non-ionic surfactants commonly contribute to the enzymatic digestion of lignocelluloses, but there are doubts about whether they are beneficial or harmful in the pretreatment and ethanol fermentation steps. This study evaluated Tween 80 (polysorbate 80) in pretreatment, enzymatic hydrolysis, and fermentation with Saccharomyces cerevisiae PE2 to obtain ethanol from green coconut fiber (GCF). To reduce the GCF’s recalcitrance and to increase enzyme accessibility, the material was pretreated using steam explosion (severity factor of 3.71 and 4.24), alkaline (sodium hydroxide), and sequential steam explosion/alkaline pretreatment in the presence of the surfactant. The surfactant did not increase delignification in steam explosion pretreatment, but it increased the sugar release when compared to the pretreatments without Tween 80. The addition of the surfactant during enzymatic hydrolysis increased glucose production in all experiments. Adsorption experiments showed that Tween 80 increased enzymatic activity. The highest glucose production was 20.31 ± 0.48 g/L and cellulosic conversion of 65.12% after adding 1.0% (w/v) Tween 80 in sequential steam explosion/alkaline pretreatment and enzymatic hydrolysis. Simultaneous saccharification and fermentation (SSF) and semi-simultaneous saccharification and fermentation (SSSF) strategies were investigated with the surfactant addition. When 2.0% (w/v) Tween 80 was added in all fermentations, it hindered ethanol production. A fed-batch SSSF and nonisothermal-fed-batch-SSSF were also performed, along with an enzyme fed-batch strategy. The highest ethanol concentration achieved was 48.21 ± 1.13 g/L by adding 1.0% (w/v) Tween 80 to the nonisothermal-fed-SSSF with 30% (w/v) solid loading.
Graphical Abstract
... are yeasts predominantly found in Arctic and Antarctic regions. Tsuji et al. (2013Tsuji et al. ( , 2014 isolated and characterized Mrakia blollopis SK-4 from Nagaike Lake in Skarvsnes ice-free area (East Antarctica) with the ability to ferment various sugars substrates like glucose, sucrose, maltose, raffinose, and fructose at low temperatures. ...
... Cold active/adapted lipase can be applied to a variety of biotechnological process, including food and beverage industries (Kavitha, 2016), decomposition of milk fat (Tsuji et al., 2015), wastewater treatment in temperate regions (Tsuji et al., 2013), detergent formulation (Sahay and Chouhan, 2018), and ethanol fermentation (Tsuji et al., 2014). Among the Antarctic yeast reported as lipase producers are: Candida antarctica Yasuda and Yamamoto, 2020), Candida zeylanoides, Vishniacozyma victoriae (formerly Cryptococcus victoriae), Leucosporidium creatinivorum (Vaz et al., 2011), Mrakia blollopsis (Tsuji et al., 2013), Leucosporidium scottii (Duarte et al., 2015), Phenoliferia glacialis, Mrakia frigida, Pichia caribbica, Leucosporidium muscorum (Martorell et al., 2017), and Rhodotorula sp. ...
The search for microbial enzymes from extreme environments may result in the discovery of biomolecules with different properties. In this sense, the aim of this study was to evaluate and optimize the conditions for lipase production by the marine-derived Antarctic yeast Leucosporidium scottii CRM 728 using an experimental design with different sources of carbon and nitrogen. The applied strategy was composed of three steps, including Plackett-Burman, Central Composite Design (CCD), and validation assay. Among nine variables applied, corn steep liquor, olive oil, and soybean oil showed a positive effect in the lipase production, while urea showed a negative effect. The production of lipase increased 9-fold in comparison to the basal activity when olive oil and corn steep liquor were used as nutrient sources. On the other hand, there was an increase of 4.8-fold when soybean oil and corn steep liquor were used as nutrient sources. The highest amounts of lipase were achieved in non-saline conditions and at 20.0 ºC. In this study, the lipase production by the marine-derived psychrotolerant yeast L. scottii was for the first time reported, providing new knowledgement in the field of enzymatic production by extremophilic and marine microbial resource.
... Moreover, previous literature indicated that supplementing Tween 80 presented positive influence on the enzymatic hydrolysis and SSF fermentation process [33]. Several studies of combining Tween 80 with SSF had been applied to increase ethanol production [25,34]; however, the comparison of three modes of batch, fed-batch, and fed-batch + Tween 80 at high solids loading with SSF has rarely been reported. ...
Background:
The mixed-feedstock fermentation is a promising approach to enhancing the co-generation of cellulosic ethanol and methane from sugarcane bagasse (SCB) and molasses. However, the unmatched supply of the SCB and molasses remains a main obstacle built upon binary feedstock. Here, we propose a cellulose-starch-sugar ternary waste combinatory approach to overcome this bottleneck by integrating the starch-rich waste of Dioscorea composita Hemls. extracted residue (DER) in mixed fermentation.
Results:
The substrates of the pretreated SCB, DER and molasses with varying ratios were conducted at a relatively low solids loading of 12%, and the optimal mixture ratio of 1:0.5:0.5 for the pretreated SCB/DER/molasses was determined by evaluating the ethanol concentration and yield. Nevertheless, it was found that the ethanol yield decreased from 79.19 ± 0.20 to 62.31 ± 0.61% when the solids loading increased from 12 to 44% in batch modes, regardless of the fact that the co-fermentation of three-component feedstock was performed under the optimal condition defined above. Hence, different fermentation processes such as fed-batch and fed-batch + Tween 80 were implemented to further improve the ethanol concentration and yield at higher solids loading ranging between 36 and 44%. The highest ethanol concentration of 91.82 ± 0.86 g/L (69.33 ± 0.46% of theoretical yield) was obtained with fed-batch + Tween 80 mode during the simultaneous saccharification and fermentation at a high solids loading of 44%. Moreover, after the ethanol recovery, the remaining stillage was digested for biomethane production and finally yielded 320.72 ± 6.98 mL/g of volatile solids.
Conclusions:
Integrated DER into the combination of SCB and molasses would be beneficial for ethanol production. The co-generation of bioethanol and biomethane by mixed cellulose-starch-sugar waste turns out to be a sustainable solution to improve the overall efficacy in biorefinery.
... Since a nonionic surfactant is thought to combine with lignin (Eriksson et al. 2002), this unproductive fermentation of eucalyptus was thought to be potentially improved by the addition of the nonionic surfactant. In fact, the ethanol concentration was increased 1.6-fold compared to the ethanol concentration in the DEF solution not containing the nonionic surfactant (Tsuji et al. 2014b). ...
Basidiomycetous yeast species of the genus Mrakia have been reported from a variety of extreme cold environments such as polar regions, glaciers, and high mountains. Several reports indicate that fungal species within the genus Mrakia occupy a major mycoflora in Antarctic environments. These results strongly suggest that this genus is well adapted to the polar environment. The genus Mrakia has unique characteristics such as an ethanol fermentation ability, and the ability to decompose milk fat under low-temperature conditions. Thus, the genus Mrakia has quite interesting characteristics. We believe that the results obtained in previous studies will contribute to the progress of related research fields and hope that further investigation will offer many opportunities to obtain more valuable knowledge on Antarctic microbes and their potential uses for human activities. In this chapter, we review the taxonomic history, physiology, ecological role, and biotechnological applications of basidiomycetous yeasts within the genus Mrakia. In the near future, this genus will become an important agent in the field of low-temperature microbiology.
... Abundant fungi present in Cu and CuTriQAC treated wood of NG were mainly mould fungi and yeasts with a capability to degrade wood (Horvath et al., 1976;Fryar et al., 2004;Bridžiuvienė and Raudoniene, 2013;Tsuji et al., 2014;Kwaśna et al., 2016), which have been found previously in preservative treated wood for members of the genera Alternaria (Bridžiuvienė and Raudoniene, 2013;Råberg et al., 2013;Kirker et al., 2017), Aureobasidium (Horvath et al., 1976), Pochonia (Kirker et al., 2017) and the family Nectriaceae (Råberg et al., 2013;Kirker et al., 2017). Fungal community compositions in CG differed from those of NG (Fig. 2) and the majority of abundant members of both community compositions were similarly known as decomposers of wood components (Deacon et al., 2006;Råberg et al., 2013). ...
Copper-based preservatives are used for wood protection, which leads to an enrichment of copper tolerant microbial communities in respective soil environments. In this study impregnated pine wood specimens (2 × 2 × 36 cm) were half buried in two field sites with (North Germany, NG) and without (Central Germany, CG) a long preservative history and incubated for 17 and 36 weeks. Pine sapwood specimens were impregnated with either CuTriQAC (containing copper, triazoles and quaternary ammonium compounds), Cu (containing only copper) or water (H2O) as control. The effect of preservative treatment at the wood-soil interface, 1 mm and 7 mm into wood was assessed by quantitative PCR and amplicon sequencing of the fungal ITS region and bacterial 16S rRNA gene of both field sites over time. Fungal and bacterial copy numbers decreased in 1 mm and only fungal in 7 mm into CuTriQAC treated wood in both sites but not in Cu treated wood. Both Cu and CuTriQAC treated wood shifted the richness, evenness of the bacterial and, to a lesser extent, the fungal community composition inside wood. In conclusion, Cu and CuTriQAC treated wood caused a selection towards microfungal ascomycetes and defined bacterial genera, which majority of genera were previously described as diazotrophs.
... En este estudio se comparó la hidrólisis y fermentación llevada a cabo por separado y de forma simultánea, obteniendo un rendimiento de producción de EAB de 15.29 g/l y 13.41 g/l, respectivamente. Otros estudios acerca de la producción de etanol a partir de residuos forestales y de madera presentan rendimientos de producción entre 15 g/l y 16 g/l ( Kaushal, Sharma y Dogra, 2016;Tsuji et al., 2014). En contraste, Wang, Zhu, Zalesny y Chen (2012) obtuvieron un rendimiento de producción de etanol de 29.7 g/l; en este trabajo determinaron que para mejorar la eficiencia en la producción de etanol es muy importante aplicar un pretratamiento eficiente de los residuos forestales. ...
Una de las prioridades del mundo actual es la generación de bioenergéticos sustentables y amigables con el medio ambiente, para lo cual son necesarios cambios en los modelos de producción y utilización de energía. El uso de residuos forestales como una fuente de biomasa para la generación de bioenergéticos representa una alternativa potencial, ya que genera un biocombustible menos contaminante en comparación con los de origen fósil. Además, mientras que el calor y la electricidad pueden ser generados por otras alternativas renovables como viento, sol y agua, la producción de combustibles y la síntesis de productos químicos requieren transformar la biomasa. La elección de la estrategia de conversión depende del tipo, propiedades y cantidad de la biomasa disponible, de los requerimientos de uso, así como de los estándares ambientales y condiciones económicas. El objetivo del presente trabajo es describir los diferentes procesos a través de los cuales se puede obtener energía a partir de residuos de biomasa forestal, así como el potencial que tienen estos residuos en la producción de los diferentes tipos de bioenergéticos. Asimismo, se describen los desafíos y problemas que aún siguen sin resolver, como lo son la recolección de residuos, escalamiento y costos de producción.
... The fermentation aptitude of some Mrakia strains was underlined in several studies that consider different scientific areas such as: i) production of beer using home brewing kit (Thomas-Hall et al., 2010), ii) ability to ferment different sugars and to produce lipase during waste water treatment (Tsuji et al., 2013), iii) production of ethanol from lignocellulosic biomass (Tsuji et al., 2014), iv) activity of milk fat decomposition (Tsuji et al., 2015), v) studies concerning the yeasts adaptability to cold (Tsuji, 2016). Although the fermentation aptitude of Mrakia spp. ...
Due to the increasing consumer demand, the production of low alcoholic and non alcoholic beer is the new goal of the present brewing producers. Although the beer with reduced alcohol content is currently obtained by physical methods, the use of non-Saccharomyces yeast, with low fermentations capacities, may represent an interesting biological approach. In this study the ethanol content and the volatile profile of a beer obtained using the basidiomycetous psychrophilic yeast strain Mrakia gelida DBVPG 5952 was compared with that produced by a commercial starter for low alcohol beers, Saccharomycodes ludwigii WSL17. The two beers were characterized by a low alcohol content (1.40% and 1.32% v/v) and by a low diacetyl production (5.04 and 5.20 μg/L). However, the organoleptic characteristics of the beer obtained using M. gelida are more appreciated by the panelists, in comparison to the analogous produced with the commercial strain of S. ludwigii.
... However, little is known about the ecological roles of Antarctic fungi inhabiting the area around Syowa Station, and further studies are therefore needed to elucidate them. In Japan, Antarctic fungi have not been Mrakia blollopis Thomas-Hall soil and lake sediment Skarvsnes ice-free area M, P AB630315, AB630320, AB763554, AB774466, AB774467, AB775474, AB908114, AB908115, AB908116, AB908117, AB908118, AB908119, AB908120, AB908121, AB908122, AB908123, AB908124, AB908125, AB908126, AB908127, AB908128, AB908178 Tsuji et al., 2013aTsuji et al., , 2013bTsuji et al., , 2013cTsuji et al., , 2013dTsuji et al., , 2014aTsuji et al., , 2014bTsuji et al., , 2015Tsuji et al., , 2016Tsuji, 2016 Mrakia gelida M indicates that the morphology of the species was reported in a previous study. P indicates that physiological characteristics were reported in a previous study. ...
The Japanese Antarctic Research Expedition (JARE) was started in 1957. The expedition marked its 60th anniversary in January 2017. In total 76 fungal species (61 ascomycetous fungi, including 9 unidentified species, and 16 basidiomycetous fungi) have thus far been recorded from the area around Syowa Station. In this review, I present a catalog of the fungal species isolated from the vicinity of Syowa Station to mark the 60th anniversary of JARE.
... Filamentous fungi recovered from Antarctic samples that belong to the genera Cadophora Cladosporium, Geomyces, Penicillium, Pseudeurotium, Pseudogymnoascus, Verticillium, and Wardomyces have shown the ability to produce cellulases [7,66,89,90]. The production of these enzymes was also observed in Antarctic yeasts belonging to the genera Cryptococcus and Mrakia [7,13,33,94]. Additionally, Penicillium roqueforti, Cadophora malorum, Geomyces sp., and Mrakia blollopis can grow and use cellulose substrates in temperatures ranging from 4.0 to 22.0 C [13,89,90]. ...
Antarctica is the coldest, windiest, and driest continent on Earth. In this sense, microorganisms that inhabit Antarctica environments have to be adapted to harsh conditions. Fungal strains affiliated with Ascomycota and Basidiomycota phyla have been recovered from terrestrial and marine Antarctic samples. They have been used for the bioprospecting of molecules, such as enzymes. Many reports have shown that these microorganisms produce cold-adapted enzymes at low or mild temperatures, including hydrolases (e.g. α-amylase, cellulase, chitinase, glucosidase, invertase, lipase, pectinase, phytase, protease, subtilase, tannase, and xylanase) and oxidoreductases (laccase and superoxide dismutase). Most of these enzymes are extracellular and their production in the laboratory has been carried out mainly under submerged culture conditions. Several studies showed that the cold-adapted enzymes exhibit a wide range in optimal pH (1.0–9.0) and temperature (10.0–70.0 °C). A myriad of methods have been applied for cold-adapted enzyme purification, resulting in purification factors and yields ranging from 1.70 to 1568.00-fold and 0.60 to 86.20%, respectively. Additionally, some fungal cold-adapted enzymes have been cloned and expressed in host organisms. Considering the enzyme-producing ability of microorganisms and the properties of cold-adapted enzymes, fungi recovered from Antarctic environments could be a prolific genetic resource for biotechnological processes (industrial and environmental) carried out at low or mild temperatures.
