A Multispecies Fungal Biofilm Approach to Enhance the Celluloyltic Efficiency of Membrane Reactors for Consolidated Bioprocessing of Plant Biomass

The constraints and advantages in cellulolytic enzymes production by fungal biofilms for a consolidated bioconversion process were investigated during this study. The biofilm cultivations were carried out in reactors designed for consolidated bioprocessing Multispecies Biofilm Membrane reactors, (MBM) where an aerobic fungal biofilm produces the lignocellulolytic enzymes while a fermenting microorganism forms the fermentation product at anaerobic conditions. It was shown that although mycelial growth was limited in the MBM reactors compared to submerged cultivations, the secretion of cellulolytic enzymes per cell dry weight was higher. When Trichoderma reesei was used as the sole enzyme producer, cellobiose accumulated in the liquid medium as the result of the deficiency of β-glucosidase in the fungal secretome. To enhance β-glucosidase activity, T. reesei was co-cultivated with A. phoenicis which is a β-glucosidase overproducer. The two fungi formed a multispecies biofilm which produced a balanced cellulolytic cocktail for the saccharification of plant biomass. The mixed biofilm reached a 2.5 fold increase in β-glucosidase production, compared to the single T. reesei biofilm. The enzymatic systems of single and mixed biofilms were evaluated regarding their efficiency on cellulosic substrates degradation. Washed solids from steam pretreated beechwood, as well as microcrystalline cellulose were used as the substrates. The enzymatic system of the multispecies biofilm released four times more glucose than the enzymatic system of T. reesei alone from both substrates and hydrolyzed 78 and 60% of the cellulose content of washed solids from beechwood and microcrystalline cellulose, respectively

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

Fermentation performance and physiology of two strains of Saccharomyces cerevisiae during growth in high gravity spruce hydrolysate and spent sulphite liquor

Background: Lignocellulosic materials are a diverse group of substrates that are generally scarce in nutrients, which compromises the tolerance and fermentation performance of the fermenting organism. The problem is exacerbated by harsh pre-treatment, which introduces sugars and substances inhibitory to yeast metabolism. This study compares the fermentation behaviours of two yeast strains using different types of lignocellulosic substrates; high gravity dilute acid spruce hydrolysate (SH) and spent sulphite liquor (SSL), in the absence and presence of yeast extract. To this end, the fermentation performance, energy status and fermentation capacity of the strains were measured under different growth conditions. Results: Nutrient supplementation with yeast extract increased sugar uptake, cell growth and ethanol production in all tested fermentation conditions, but had little or no effect on the energy status, irrespective of media. Nutrient-supplemented medium enhanced the fermentation capacity of harvested cells, indicating that cell viability and reusability was increased by nutrient addition. Conclusions: Although both substrates belong to the lignocellulosic spruce hydrolysates, their differences offer specific challenges and the overall yields and productivities largely depend on choice of fermenting strain

Toward a sustainable biorefinery using high-gravity technology

The realization of process solutions for a sustainable bioeconomy depends on the efficient processing of biomass. High-gravity technology is one important alternative to realizing such solutions. The aims of this work were to expand the knowledge-base on lignocellulosic bioconversion processes at high solids content, to advance the current technologies for production of second-generation liquid biofuels, to evaluate the environmental impact of the proposed process by using life cycle assessment (LCA), and to develop and present a technically, economically, and environmentally sound process at high gravity, i.e., a process operating at the highest possible concentrations of raw material. The results and opinions presented here are the result of a Nordic collaborative study within the framework of the HG Biofuels project. Processes with bioethanol or biobutanol as target products were studied using wheat straw and spruce as interesting Nordic raw materials. During the project, the main scientific, economic, and technical challenges of such a process were identified. Integrated solutions to these challenges were proposed and tested experimentally, using wheat straw and spruce wood at a dry matter content of 30% (w/w) as model substrates. The LCA performed revealed the environmental impact of each of the process steps, highlighting the importance of the enzyme dose used for the hydrolysis of the plant biomass, as well as the importance of the fermentation yield

Biotechnological potential of brewers spent grain and its recent applications

Purpose Brewers spent grain (BSG) is a by-product of the brewing process corresponding to around 85% of total by-products generated. The great number of publications over the last 5 years, on the biotechnological applications of BSG, represents the increased scientific interest on it. This critical, state of the art review aims at gathering and analysing the most recent scientific efforts on the biotechnological potential of Brewer's spent grain and on its evaluation as a feedstock for high added value products. Methods The assiduous bibliographic retrospection focused on the latest scientific reports. The consideration of all relevant scientific articles was thorough and critical. The classification of the scientific efforts was made not only according to the end-products but also according to the biotechnological approach adopted. Results BSG has been used in a wide range of biotechnological applications such as substrate for enzymes production, as a source for value-added products (antioxidants, monosaccharides, oligosaccharides, xylitol, arabitol, bioethanol, biogas or lactic acid) or for the production of functional proteins and lipids. Its applications as a carrier in various bioprocesses have also been reported. Conclusion The implementation of BSG's fractionation in industrial scale seems to be the next step in BSG's exploitation. A fractionation process which allows the exploitation of biomolecules belonging to different classes, produced from one feedstock (BSG) may be used as a pattern for the implementation of the biorefinery concept in industrial scale, as long as the methods adopted ensure the functionality of the potentially valuable components

Evaluation of different strategies to overcome the inhibitory effects at high gravity processes using multivariate data analysis (MVDA)

High-gravity (HG) technology aims at generating final ethanol concentrations above 50 kg m3 in order to reduce the cost of the distillation step. The generation of higher amounts of inhibitors during the pretreatment step is one of the challenges that accom-pany the increase in initial dry matter. Detoxification of spruce hydrolysate, adaptation of the cells before fermentation, supplementation with nutrients, and washing of solids were the strategies compared in this study. They represent different approaches tocope with the inhibitory effects, and we compared their efficiencies using a thermotolerant strain of Saccharomyces cerevisiae at temperatures from 30oC up to 40oC.The dilute acid-pretreated spruce used as substrate in this study was not fermentable under HG conditions (200 g kg-1water-insoluble solids) when no improvement method was applied. In HG simultaneous saccharification and fermentationat 30oC combined with a 24 h pre-hydrolysis step, the detoxification of pretreated spruce with reducing agent (Na2S2O4) gave the best result with an ethanol yield of 57% (on total sugars) of the maximum theoretical and a volumetric productivity of1.58 g dm3 h−1. In HG separate hydrolysis and fermentation, nutrients supplementation gave better final ethanol yields than detoxification of the material, reaching an ethanol yield of about 60% of the theoretical (on total sugars). The results obtained, showed an increase in severity of inhibitory effects with temperature increase. Improved cell viability was observed when detoxified material was used and also when yeast extract addition was coupled with adaptation of the cells to the hydrolysate. The different fractions of hydrolysates after the application of different treatments were characterised and analysed using MVDA in order to evaluate the differences observed