Enhanced ethanol production from brewer's spent grain by a Fusarium oxysporum consolidated system

<p>Abstract</p> <p>Background</p> <p>Brewer's spent grain (BG), a by-product of the brewing process, is attracting increasing scientific interest as a low-cost feedstock for many biotechnological applications. BG in the present study is evaluated as a substrate for lignocellulolytic enzyme production and for the production of ethanol by the mesophilic fungus <it>Fusarium oxysporum </it>under submerged conditions, implementing a consolidated bioconversion process. Fermentation experiments were performed with sugar mixtures simulating the carbohydrate content of BG in order to determine the utilization pattern that could be expected during the fermentation of the cellulose and hemicellulose hydrolysate of BG. The sugar mixture fermentation study focused on the effect of the initial total sugar concentration and on the effect of the aeration rate on fermenting performance of <it>F. oxysporum</it>. The alkali pretreatment of BG and different aeration levels during the ethanol production stage were studied for the optimization of the ethanol production by <it>F. oxysporum</it>.</p> <p>Results</p> <p>Enzyme yields as high as 550, 22.5, 6.5, 3225, 0.3, 1.25 and 3 U per g of carbon source of endoglucanase, cellobiohydrolase, β-D-glucosidase, xylanase, feruloyl esterase, β-D-xylosidase and α-L-arabinofuranosidase respectively, were obtained during the growth stage under optimized submerged conditions. An ethanol yield of 109 g ethanol per kg of dry BG was obtained with alkali-pretreated BG under microaerobic conditions (0.01 vvm), corresponding to 60% of the theoretical yield based on total glucose and xylose content of BG.</p> <p>Conclusion</p> <p>The enzymatic profile of the extracellular extract from <it>F. oxysporum </it>submerged cultures using BG and corn cob as the carbon source was proved efficient for a successful hydrolysis of BG. The fermentation study carried out using sugar mixtures simulating BG's carbohydrates content and consecutively alkali-pretreated and untreated BG, indicates that BG hydrolysis is the bottleneck of the bioconversion process. However, a considerable bioconversion yield was achieved (60% of the theoretical) making this bioprocess worthy of further investigation for a potential commercial application.</p

Development of Thermophilic Tailor-Made Enzyme Mixtures for the Bioconversion of Agricultural and Forest Residues

Even though the main components of all lignocellulosic feedstocks include cellulose, hemicellulose, as well as the protective lignin matrix, there are some differences in structure, such as in hardwoods and softwoods, which may influence the degradability of the materials. Under this view, various types of biomass might require a minimal set of enzymes that has to be tailor-made. Partially defined complex mixtures that are currently commercially used are not adapted to efficiently degrade different materials, so novel enzyme mixtures have to be customized. Development of these cocktails requires better knowledge about the specific activities involved, in order to optimize hydrolysis. The role of filamentous fungus Myceliophthora thermophila and its complete enzymatic repertoire for the bioconversion of complex carbohydrates has been widely proven. In this study, four core cellulases (MtCBH7, MtCBH6, MtEG5 and MtEG7), in the presence of other four accessory enzymes (mannanase, lytic polyssacharide monooxygenase MtGH61, xylanase, MtFae1a) and β-glucosidase MtBGL3, were tested as a 9-component cocktail against one model substrate (phosphoric acid swollen cellulose) and four hydrothermally pretreated natural substrates (wheat straw as an agricultural waste, birch and spruce biomass, as forest residues). Synergistic interactions among different enzymes were determined using a suitable design of experiments methodology. The results suggest that for the hydrolysis of the pure substrate (PASC), high proportions of MtEG7 are needed for efficient yields. MtCBH7 and MtEG7 are enzymes of major importance during the hydrolysis of pretreated wheat straw, while MtCBH7 plays a crucial role in case of spruce. Cellobiohydrolases MtCBH6 and MtCBH7 act in combination and are key enzymes for the hydrolysis of the hardwood (birch). Optimum combinations were predicted from suitable statistical models which were able to further increase hydrolysis yields, suggesting that tailor-made enzyme mixtures targeted towards a particular residual biomass can help maximize hydrolysis yields. The present work demonstrates the change from ‘one cocktail for all’ to ‘tailor-made cocktails’ that are needed for the efficient saccharification of targeted feed stocks prior to the production of biobased products through the biorefinery concept

Carbonic anhydrase to boost CO2 sequestration : Improving carbon capture utilization and storage (CCUS)

CO2 Capture Utilization and Storage (CCUS) is a fundamental strategy to mitigate climate change, and carbon sequestration, through absorption, can be one of the solutions to achieving this goal. In nature, carbonic anhydrase (CA) catalyzes the CO2 hydration to bicarbonates. Targeting the development of novel biotechnological routes which can compete with traditional CO2 absorption methods, CA utilization has presented a potential to expand as a promising catalyst for CCUS applications. Driven by this feature, the search for novel CAs as biocatalysts and the utilization of enzyme improvement techniques, such as protein engineering and immobilization methods, has resulted in suitable variants able to catalyze CO2 absorption at relevant industrial conditions. Limitations related to enzyme recovery and recyclability are still a concern in the field, affecting cost efficiency. Under different absorption approaches, CA enhances both kinetics and CO2 absorption yields, besides reduced energy consumption. However, efforts directed to process optimization and demonstrative plants are still limited. A recent topic with great potential for development is the CA utilization in accelerated weathering, where industrial residues could be re-purposed towards becoming carbon sequestrating agents. Furthermore, research of new solvents has identified potential candidates for integration with CA in CO2 capture, and through techno-economic assessments, CA can be a path to increase the competitiveness of alternative CO2 absorption systems, offering lower environmental costs. This review provides a favorable scenario combining the enzyme and CO2 capture, with possibilities in reaching an industrial-like stage in the future

High concentrations of dried sorghum stalks as a biomass feedstock for single cell oil production by Rhodosporidium toruloides

Environmental crisis and concerns for energy security have made the research for renewable fuels that will substitute the usage of fossil fuels an important priority. Biodiesel is a potential substitute for petroleum, but its feasibility is hindered by the utilization of edible vegetable oil as raw material, which is responsible for a large fraction of the production cost and fosters the food versus fuel competition. Microbial oils are an interesting alternative as they do not compete with food production, and low cost renewable materials could serve as raw materials during cultivation of microorganisms. Sweet sorghum is an excellent candidate as substrate for microbial oil production, as it possesses high photosynthetic activity yielding high amounts of soluble and insoluble carbohydrates, and does not require high fertilization and irrigation rates8FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informaçã

Characterization of Organosolv Birch Lignins: Toward Application-Specific Lignin Production

Organosolv pretreatment represents one of the most promising biomass valorization strategies for renewable carbon-based products; meanwhile, there is an overall lack of holistic approach to how extraction conditions affect the suitable end-usages. In this context, lignin extracted from silver birch (Betula pendula L.) by a novel hybrid organosolv/steam-explosion treatment at varying process conditions (EtOH %; time; catalyst %) was analyzed by quantitative NMR (1H-13C HSQC; 13C NMR; 31P NMR), gel permeation chromatography, Fourier transform infrared (FT-IR), Pyr-gas chromatography-mass spectroscopy (GC/MS), and thermogravimetric analysis, and the physicochemical characteristics of the lignins were discussed regarding their potential usages. Characteristic lignin interunit bonding motifs, such as beta-O-4', beta-beta', and beta-5', were found to dominate in the extracted lignins, with their abundance varying with treatment conditions. Low-molecular-weight lignins with fairly unaltered characteristics were generated via extraction with the highest ethanol content potentially suitable for subsequent production of free phenolics. Furthermore, beta-beta' and beta-5' structures were predominant at higher acid catalyst contents and prolonged treatment times. Higher acid catalyst content led to oxidation and ethoxylation of side-chains, with the concomitant gradual disappearance of p-hydroxycinnamyl alcohol and cinnamaldehyde. This said, the increasing application of acid generated a broad set of lignin characteristics with potential applications such as antioxidants, carbon fiber, nanoparticles, and water remediation purposes

Prediction of yields and composition of char from fast pyrolysis of commercial lignocellulosic materials, organosolv fractionated and torrefied olive stones

peer-reviewedThis study investigated the fast pyrolysis behaviour of torrefied olive stones, fractionated olive stones and lignocellulosic commercial compounds. Olive stones were reacted in a continuous industrial torrefaction unit. The olive stones were also fractionated into their main components in an organosolv reactor at temperatures from 170 to 190 ◦C in both the presence and absence of an acidic catalyst. All samples were reacted in a wire mesh reactor at different temperatures (800–1150 ◦C) and heating rates (400–1150 ◦C/s), and the solid product was characterised for its yield, morphology, and elemental composition. The char yields from fast pyrolysis of commercially available cellulose, hemicelluloses, and lignin were compared with yields of fractionated olive stones. A model was developed to compare the measured yields of olive stones with the predicted yields using fractionated or commercial components. The presence of acid during fractionation had a stronger effect than the temperature, particularly on the lignin fraction. The fractionated lignocellulosic compounds provided more accurate predictions of the char yields of olive stones, as compared to the commercial lignocellulosic compounds. The fractionation at 180 ◦C without acid catalyst gave the cellulose, hemicellulose, and lignin with highest degree of purity and resulted in the most accurate predictions of the experimental yields of olive stones. The results showed that interactions between the lignocellulosic components were not significant. The char yield of each fractioned compound and non-treated olive stones could be accurately predicted from the lignocellulosic content which has importance for biorefinery applications in which each fraction is used as a value-added product

Green conversion of municipal solid wastes into fuels and chemicals

Presently, the society is facing a serious challenge for the effective management of the increasing amount of produced municipal solid wastes. The accumulated waste has caused a series of environmental problems such as uncontrolled release of greenhouse gases. Moreover, the increasing amount of wastes has resulted in a shortage of areas available for waste disposal, resulting in a nonsustainable waste management. These problems led to serious public concerns, which in turn resulted in political actions aiming to reduce the amount of wastes reaching the environment. These actions aim to promote sustainable waste management solutions. The main objective of these policies is to promote the recycling of municipal solid waste and the conversion of waste to energy and valuable chemicals. These conversions can be performed using either biological (e.g., anaerobic digestion) or thermochemical processes (e.g., pyrolysis). Research efforts during the last years have been fruitful, and many publications demonstrated the effective conversation of municipal solid waste to energy and chemicals. These processes are discussed in the current review article together with the change of the waste policy that was implemented in the EU during the last years

Isolation and modification of nano-scale cellulose from organosolv-treated birch through the synergistic activity of LPMO and endoglucanases

Nanocellulose isolation fromlignocellulose is a tedious and expensive processwith high energy and harsh chemical requirements, primarily due to the recalcitrance of the substrate, which otherwise would have been costeffective due to its abundance. Replacing the chemical steps with biocatalytic processes offers opportunities to solve this bottleneck to a certain extent due to the enzymes substrate specificity and mild reaction chemistry. In this work, we demonstrate the isolation of sulphate-free nanocellulose from organosolv pretreated birch biomass using different glycosyl-hydrolases, along with accessory oxidative enzymes including a lytic polysaccharide monooxygenase (LPMO). The suggested process produced colloidal nanocellulose suspensions (zeta-potential-19.4 mV) with particles of 7-20 nm diameter, high carboxylate content and improved thermostability (T-o= 301 degrees C, T-max= 337 degrees C). Nanocelluloseswere subjected to post-modification using LPMOs of different regioselectivity. The sample from chemical route was the least favorable for LPMO to enhance the carboxylate content, while that from the C1-specific LPMO treatment showed the highest increase in carboxylate content. (c) 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/)

optimization of enzymatic synthesis of l arabinose ferulate catalyzed by feruloyl esterases from myceliophthora thermophila in detergentless microemulsions and assessment of its antioxidant and cytotoxicity activities

The feruloyl esterases FaeA1, FaeA2, FaeB1, FaeB2 from Myceliophthora thermophila C1 and MtFae1a from M. thermophila ATCC 42464 were used as biocatalysts for the transesterification of vinyl ferula ..