Challenges and prospects of yeast-based microbial oil production within a biorefinery concept.

Biodiesel, unlike to its fossil-based homologue (diesel), is renewable. Its use contributes to greater sustainability in the energy sector, mainly by reducing greenhouse gas emissions. Current biodiesel production relies on plant- and animal-related feedstocks, resulting in high final costs to the prices of those raw materials. In addition, the production of those materials competes for arable land and has provoked a heated debate involving their use food vs. fuel. As an alternative, single-cell oils (SCOs) obtained from oleaginous microorganisms are attractive sources as a biofuel precursor due to their high lipid content, and composition similar to vegetable oils and animal fats. To make SCOs competitive from an economic point of view, the use of readily available low-cost substrates becomes essential. This work reviews the most recent advances in microbial oil production from non-synthetic sugar-rich media, particularly sugars from lignocellulosic wastes, highlighting the main challenges and prospects for deploying this technology fully in the framework of a Biorefinery concept.

Efficient use of discarded vegetal residues as cost-effective feedstocks for microbial oil production.

BACKGROUND: Horticultural intensive type systems dedicated in producing greenhouse vegetables are one of the primary industries generating organic waste. Towards the implementation of a zero-waste strategy, this work aims to use discarded vegetables (tomato, pepper and watermelon) as feedstock for producing microbial oil using the oleaginous yeast Cryptococcus curvatus. RESULTS: The soluble fraction, resulting after crushing and centrifuging these residues, showed C/N ratios of about 15, with a total carbohydrate content (mainly glucose, fructose and sucrose) ranging from 30 g/L to 65 g/L. Using these liquid fractions as substrate under a pulse-feeding strategy with a concentrated glucose solution resulted in an intracellular total lipid accumulation of about 30% (w/w) of the total dry cell weight (DCW). To increase this intracellular lipid content, the initial C/N content was increased from 15 to 30 and 50. Under these conditions, the process performance of the pulse-feeding strategy increased by 20-36%, resulting in a total intracellular lipid concentration of 35-40% DCW (w/w). CONCLUSION: These results demonstrate the potential of discarded vegetables as a substrate for producing bio-based products such as microbial oil when proper cultivation strategies are available.

Recent advances on physical technologies for the pretreatment of food waste and lignocellulosic residues.

The complete deployment of a bio-based economy is essential to meet the United Nations' Sustainable Development Goals from the 2030 Agenda. In this context, food waste and lignocellulosic residues are considered low-cost feedstocks for obtaining industrially attractive products through biological processes. The effective conversion of these raw materials is, however, still challenging, since they are recalcitrant to bioprocessing and must be first treated to alter their physicochemical properties and ease the accessibility to their structural components. Among the full pallet of pretreatments, physical methods are recognised to have a high potential to transform food waste and lignocellulosic residues. This review provides a critical discussion about the recent advances on milling, extrusion, ultrasound, and microwave pretreatments. Their mechanisms and modes of application are analysed and the main drawbacks and limitations for their use at an industrial scale are discussed.

In Vitro Assessment of the Prebiotic Potential of Xylooligosaccharides from Barley Straw.

Barley straw was subjected to hydrothermal pretreatment (steam explosion) processing to evaluate its potential as a raw material to produce xylooligosaccharides (XOS) suitable for use as a prebiotic. The steam explosion pretreatment generated a liquid fraction containing solubilised hemicellulose. This fraction was purified using gel permeation chromatography to obtain a fraction rich in XOS DP2-DP6. The sample was characterised through analytical techniques such as HPAEC-PAD, FTIR and MALDI-TOF-MS. The prebiotic activity was evaluated using in vitro fermentation in human faecal cultures through the quantification of short-chain fatty acid (SCFA) and lactate production, the evolution of the pH and the consumption of carbon sources. The total SCFA production at the end of fermentation (30 h) was 90.1 mM. Positive significant differences between the amount of XOS from barley straw and fructooligosaccharides after incubation were observed.

Production of xylooligosaccharides, bioethanol, and lignin from structural components of barley straw pretreated with a steam explosion.

Barley straw (BS) is a potential source to obtain bioethanol and value-added products such as xylooligosaccharides (XOS) and lignin for application in diverse industries. In this study, BS was submitted to steam explosion pretreatment to valorize the main components of this lignocellulose biomass. For hemicellulose fraction valorization, different combinations of endo-ß-(1,4)-D-xylanase enzyme with accessory enzymes (α-L-arabinofuranosidase, feruloy -esterase and acetylxylan-esterase) have been studied to produce XOS with a low degree of polymerization. The application of accessory enzymes combined with endo-ß-(1,4)-D-xylanase enzymes turned out to be the most effective strategy for the formation of XOS. The solid fraction obtained after the pretreatment was submitted to presacharification and simultaneous saccharification and fermentation process for bioethanol production. The resulting lignin-rich residue was characterized. In this integrated process, 13.0 g XOS (DP2-DP6), 12.6 g ethanol and 16.6 g lignin were obtained from 100 g of BS, achieving the goal of valorizing this agricultural residue.

Valorization of Greenhouse Horticulture Waste from a Biorefinery Perspective.

Greenhouse cultivation and harvesting generate considerable amounts of organic waste, including vegetal waste from plants and discarded products. This study evaluated the residues derived from tomato cultivation practices in Almería (Spain) as sugar-rich raw materials for biorefineries. First, lignocellulose-based residues were subjected to an alkali-catalyzed extrusion process in a twin-screw extruder (100 °C and 6-12% (w/w) NaOH) to assess maximum sugar recovery during the subsequent enzymatic hydrolysis step. A high saccharification yield was reached when using an alkali concentration of 12% (w/w), releasing up to 81% of the initial glucan. Second, the discarded tomato residue was crushed and centrifuged to collect both the juice and the pulp fractions. The juice contained 39.4 g of sugars per 100 g of dry culled tomato, while the pulp yielded an extra 9.1 g of sugars per 100 g of dry culled tomato after an enzymatic hydrolysis process. The results presented herein show the potential of using horticulture waste as an attractive sugar source for biorefineries, including lignocellulose-based residues when effective fractionation processes, such as reactive extrusion technology, are available.

Integral process assessment of sugarcane agricultural crop residues conversion to ethanol.

This work focuses a whole process assessment on post-harvesting sugarcane residues for 2G ethanol production by different saccharification-fermentation conditions at high solids loading, performed after steam explosion, alkaline and acidic pretreatments. Carbohydrate recoveries and enzymatic digestibility results showed that alkali and steam explosion pretreatments were effective for the biomass assayed. Due to a significant improvement (60%) of the glucose released by combining hemicellulases and cellulases only after the NaOH pretreatment, the most favorable process settled comprised an alkali-based pretreatment followed by a pre-saccharification and simultaneous saccharification and fermentation (PSSF). The produced ethanol reached 4.8% (w/w) as a result of an 80% conversion of the glucose from the pretreated biomass. Finally, an ethanol concentration of 3.2% (w/w) was obtained by means of a steam explosion followed by PSSF, representing a suitable start point to further develop a low environmental impact alternative for ethanol production.

Strategies of xylanase supplementation for an efficient saccharification and cofermentation process from pretreated wheat straw.

Ethanol production from lignocellulosic raw materials includes a pretreatment step before enzymatic hydrolysis (EH). Pretreated substrates contain complex hemicelluloses in the solid fraction that can protect the cellulose from enzymatic attack. In addition, soluble xylooligomers are contained in the pretreated materials and may have an inhibitory effect on cellulase activity. In this context, several approaches for xylanase supplementation have been studied to increase EH yields. In this study, the whole slurry obtained after steam explosion pretreatment of wheat straw has been used as substrate. EH experiments were performed using commercial cellulase preparations supplemented with an endoxylanase (XlnC) from Aspergillus nidulans. Among different strategies of XlnC supplementation, the 24-h xylanase treatment before cellulase addition yielded an increase of 40.1 and 10.1% in glucose and xylose production, respectively. Different XlnC addition strategies were integrated in a simultaneous saccharification and cofermentation process (SSCF) using the xylose fermenting strain Saccharomyces cerevisiae F12. Ethanol production in SSCF was 28.4% higher when comparing to a simultaneous saccharification and fermentation process.

Ethanol production from the organic fraction obtained after thermal pretreatment of municipal solid waste.

In this work, the use of organic fraction from municipal solid waste (MSW) as substrate for ethanol production based on enzymatic hydrolysis was evaluated. MSW was subjected to a thermal pretreatment (active hygienization) at 160 degrees C from 5 to 50 min. The organic fiber obtained after 30 min was used as substrate in a simultaneous saccharification and fermentation (SSF) and fed-batch SSF process using cellulases and amylases. In a fed-batch mode with 25% (w/w) substrate loading, final ethanol concentration of 30 g/L was achieved (60% of theoretical). In these conditions, more than 160 L of ethanol per ton of dry matter could be produced from the organic fraction of MSW.

Hydrothermal pretreatment conditions to enhance ethanol production from poplar biomass.

Pretreatment has been recognized as a key step in enzyme-based conversion processes of lignocellulose biomass to ethanol. The aim of this study is to evaluate two hydrothermal pretreatments (steam explosion and liquid hot water) to enhance ethanol production from poplar (Populus nigra) biomass by a simultaneous saccharification and fermentation (SSF) process. The composition of liquid and solid fractions obtained after pretreatment, enzymatic digestibility, and ethanol production of poplar biomass pretreated at different experimental conditions was analyzed. The best results were obtained in steam explosion pretreatment at 210 C and 4 min, taking into account cellulose recovery above 95%, enzymatic hydrolysis yield of about 60%, SSF yield of 60% of theoretical, and 41% xylose recovery in the liquid fraction. Large particles can be used for poplar biomass in both pretreatments, since no significant effect of particle size on enzymatic hydrolysis and SSF was obtained.

Effect of lignocellulosic degradation compounds from steam explosion pretreatment on ethanol fermentation by thermotolerant yeast Kluyveromyces marxianus.

The filtrate from steam-pretreated poplar was analyzed to identify degradation compounds. The effect of selected compounds on growth and ethanolic fermentation of the thermotolerant yeast strain Kluyveromyces marxianus CECT 10875 was tested. Several fermentations on glucose medium, containing individual inhibitory compounds found in the hydrolysate, were carried out. The degree of inhibition on yeast strain growth and ethanolic fermentation was determined. At concentrations found in the prehy-drolysate, none of the individual compounds significantly affected the fermentation. For all tested compounds, growth was inhibited to a lesser extent than ethanol production. Lower concentrations of catechol (0.96 g/L) and 4-hydroxybenzaldehyde (1.02 g/L) were required to produce the 50% reduction in cell mass in comparison to other tested compounds.

Ethanol production from olive oil extraction residue pretreated with hot water.

The olive pulp fraction contained in the residue generated in olive oil extraction by a two-step centrifugation process can be upgraded by using the cellulose fraction to produce ethanol and recovering high value phenols (tyrosol and hydroxytyrosol). Olive pulp was pretreated in a laboratory scale stirred autoclave at different temperatures (150-250 degrees C). Pretreatment was evaluated regarding cellulose recovery, enzymatic hydrolysis effectiveness, ethanol production by a simultaneous saccharification and fermentation process (SSF), and phenols recovery in the filtrate. The pretreatment of olive pulp using water at temperatures between 200 degrees C and 250 degrees C enhanced enzymatic hydrolysis. Maximum ethanol production (11.9 g/L) was obtained after pretreating pulp at 210 degrees C in a SSF fed-batch procedure. Maximum hydroxytyrosol recovery was obtained in the liquid fraction when pretreated at 230 degrees C.