Plant Cell Wall Material Research Articles

A determination of the composition and structure of the polysaccharides fractions isolated from apple cell wall based on FT-IR and FT-Raman spectra supported by PCA analysis

Generally in plants, cell wall polysaccharides are the main determinants of tissue properties and contents of an essential part of dietary fiber, prebiotics, or cosmetic ingredients. Therefore, evaluation of the content and structure of polysaccharides is crucial to understanding and controlling their functionality. Vibrational spectroscopy has already proven to be very useful in studying wall-based compounds. Here, it was proposed to use the FT-IR and FT-Raman spectra combined with PCA analysis to discriminate and distinguish pectic and hemicellulosic fractions isolated from apple parenchyma as an example. Reference measurements have been done on commercially available polysaccharides in order to identify relevant fingerprint bands. The similarities of different polysaccharide fractions from apple: water, imidazole, dilute alkali-soluble polysaccharides (WSP, ISP, DASP, respectively), and polysaccharides soluble in three potassium hydroxide concentration (KOHs) to the reference polysaccharides were found. In the case of both types of spectroscopy, the region below 1800 cm−1 was used to differentiate the samples. The results showed that the WSP, ISP, and DASP fractions have band patterns similar to the pectic polysaccharides, but also contain bands characteristic of polysaccharides containing arabinan and galactan. While the KOH fractions showed the greatest similarities to the hemicellulose polysaccharides, especially the glucomannan and xyloglucan. The results showed that FT-IR and FT-Raman spectra supported by PCA can be useful for studying the structure of polysaccharide fractions directly isolated from plant cell wall material.

Mass transfer study on noncovalent adsorption and desorption of soluble phenolics by plant cell wall materials for green purification

This study originated from the observation that plant cell walls can bind soluble phenolics through noncovalent interactions. Celery and blueberry cell wall materials obtained from waste sources were selected as effective adsorbents for purifying phenolics. First, the impact of environmental ionic strength on the adsorption of phenolics by plant cell walls was investigated, indicating that ionic interactions play a more important role in adsorption than hydrophobic interactions. Second, numerical simulation results found that the increments of diffusivities of phenolics both within the liquids in the pores of plant cell wall materials and the skeleton of cell wall adsorbents accelerated the adsorption. The abundance of adsorption sites within plant cell wall adsorbents determined their adsorption capacity, and the penetration of phenolics across the adsorbent/liquid interface was unrelated to the adsorption behavior. Third, due to the porous structure of celery cell wall adsorbents, the pore volume diffusion of phenolics impacted intraparticle diffusion within celery cell wall materials more than blueberry cell wall materials. Fourth, after plant cell wall adsorption and desorption, the obtained phenolics achieved a purity of 60%, comparable to the purification achieved by macroporous resin adsorption/desorption. Moreover, purification using plant cell wall adsorption/desorption was more efficient. These findings contribute to the theoretical understanding of interactions between plant cell walls and polyphenols and provide valuable insights for designing natural plant cell wall adsorbents and developing environmentally friendly polyphenol purification techniques.

Identifying plant cell wall remnants in detritus of a subtropical wetland with fluorescence labeling

Sediment accretion in wetlands represents a significant carbon burial pathway. While litter studies can quantify the loss rates of plant leaf material, those studies do not provide insight into the specific cell wall polymers being retained or lost within the detrital matrix. The Everglades ecosystem has been dramatically altered due to anthropogenic eutrophication and hydrologic modifications. The results are changes in macrophyte species composition and sediment accretion- and loss- rates. To improve ecological conditions, active management strategies are re-establishing open water slough environments. A question remains about the persistence of new- and old- plant cell wall material in sediments because of active management. In this pilot project we utilized immuno-fluorescence labeling with lectins applied to plant leaf material and detrital flocculent collected from created open and control plots in the Everglades to observe the presence, absence, and overlap of specific cell wall polymers between macrophytes and detrital flocculent in increasingly recalcitrant materials that would most likely contribute to peat accumulation. The persistence and loss of specific polymers between treatment and control plots provided insight into the differing levels of recalcitrance amongst plant cell walls and their relative potential as a carbon sink. This study provides a novel method for testing for the presence and persistence of specific cell wall polymers in detritus to gain a better understanding of plant material persistence in wetland ecosystems.

The gastrointestinal microbiome of browsing goats (Capra hircus).

Despite the growing interest in the ruminants’ gastrointestinal tract (GIT) microbiomes’ ability to degrade plant materials by animal husbandry and industrial sectors, only a few studies addressed browsing ruminants. The present work describes the taxonomic and functional profile of the bacterial and archaeal communities from five different gastrointestinal sections (rumen, omasum-abomasum, jejunum, cecum and colon) of browsing Capra hircus, by metabarcoding using 16S rRNA genes hypervariable regions. The bacterial communities across the GITs are mainly composed of Bacillota and Bacteroidota. Prevotella was the leading bacterial group found in the stomachs, Romboutsia in the jejuna, and Rikenellaceae_RC9_gut_group, Bacteroides, UCG-010_ge, UCG-005, and Alistipes in large intestines. The archaeal communities in the stomachs and jejuna revealed to be mainly composed of Methanobrevibacter, while in the large intestines its dominance is shared with Methanocorpusculum. Across the GITs, the main metabolic functions were related to carbohydrate, amino acid, and energy metabolisms. Significant differences in the composition and potential biological functions of the bacterial communities were observed among stomachs, jejuna and large intestines. In contrast, significant differences were observed among stomachs and jejuna verse large intestines for archaeal communities. Overall different regions of the GIT are occupied by different microbial communities performing distinct biological functions. A high variety of glycoside hydrolases (GHs) indispensable for degrading plant cell wall materials were predicted to be present in all the GIT sections.

Multiple strategies of plant colonization by beneficial endophytic Enterobacter sp. SA187.

Although many endophytic plant growth-promoting rhizobacteria have been identified, relatively little is still known about the mechanisms by which they enter plants and promote plant growth. The beneficial endophyte Enterobacter sp. SA187 was shown to maintain the productivity of crops in extreme agricultural conditions. Here we present that roots of its natural host (Indigofera argentea), alfalfa, tomato, wheat, barley and Arabidopsis are all efficiently colonized by SA187. Detailed analysis of the colonization process in Arabidopsis showed that colonization already starts during seed germination, where seed-coat mucilage supports SA187 proliferation. The meristematic zone of growing roots attracts SA187, allowing epiphytic colonization in the elongation zone. Unlike primary roots, lateral roots are significantly less epiphytically colonized by SA187. Root endophytic colonization was found to occur by passive entry of SA187 at lateral-root bases. However, SA187 also actively penetrates the root epidermis by enzymatic disruption of plant cell wall material. In contrast to roots, endophytic colonization of shoots occurs via stomata, whereby SA187 can actively re-open stomata similarly to pathogenic bacteria. In summary, several entry strategies were identified that allow SA187 to establish itself as a beneficial endophyte in several plant species, supporting its use as a plant growth-promoting bacterium in agriculture systems.

Organically-bound silicon enhances resistance to enzymatic degradation and nanomechanical properties of rice plant cell walls

Plant cell walls exhibit excellent mechanical properties, which form the structural basis for sustainable bioresources and multifunctional nanocelluloses. The wall nanomechanical properties of living cells through covalent modifications of hybrid inorganic elements, such as silicon, may confer significant influence on local mechano-response and enzymatic degradation. Here, we present a combination of ex situ measurements of enzyme-released oligosaccharide fragments using MALDI-TOF MS and in situ atomic force microscopy (AFM) imaging through PeakForce quantitative nanomechanical mapping of tip-functionalized single-molecule enzyme-polysaccharide substrate recognition and the nanoscale dissolution kinetics of individual cellulose microfibrils of living rice (Oryza sativa) cells following silicate cross-linking of cell wall xyloglucan. We find that xyloglucan-bound silicon enhances the resistance to degradation by cellulase and improves the wall nanomechanical properties in the elastic modulus at the single-cell level. The findings establish a direct link between an inorganic element of silicon and the nanoscale architecture of plant cell wall materials for sustainable utilization.

Analytical implications of different methods for preparing plant cell wall material

Almost all plant cells are surrounded by a wall constructed of co-extensive networks of polysaccharides and proteoglycans. The capability to analyse cell wall components is essential for both understanding their complex biology and to fully exploit their numerous practical applications. Several biochemical and immunological techniques are used to analyse cell walls and in almost all cases the first step is the preparation of an alcohol insoluble residue (AIR). There is significant variation in the protocols used for AIR preparation, which can have a notable impact on the downstream extractability and detection of cell wall components. To explore these effects, we have formally compared ten AIR preparation methods and analysed polysaccharides subsequently extracted using high-performance anion exchange chromatography (HPAEC-PAD) and Micro Array Polymer Profiling (MAPP). Our results reveal the impact that AIR preparation has on downstream detection of cell wall components and the need for optimisation and consistency when preparing AIR.

Effect of flavonoid structure and pH on iron-mediated pectin interaction

Flavonoids are polyphenolic natural compounds with suggested health-promoting properties and impact on sensory aspects. Processing, as well as mastication of plant-based food, can allow interactions between flavonoids and plant cell wall material (CWM). Pectin is a cell wall polysaccharide, present in all plant-based products, and used in a wide range of foods. Recently, a novel iron-mediated mechanism for pectin-flavonoid interactions was suggested. The current work aimed to study the impact of flavonoid structure and pH on the iron-mediated interaction by using 7 aglycones (belonging to the flavonol, flavanonol, and flavone sub-groups) differing by specific small structural moieties resulting in different binding sites. By using commercial water-soluble pectin from apple pomace and an identical (except a higher content of iron) iron enriched pectin, we have qualitatively and quantitatively presented the importance of the C2–C3 double bond for such interactions. Without such a bond the aglycones do not bind to the pectin, likely due to the loss of molecular planarity. This was studied using UV-VIS spectroscopy and membrane centrifugation. Also, the binding dissociation constants (Kd) of the flavonoids presenting a specific wavelength for the ternary complexes were determined using UV-VIS spectroscopy, showing that the lack of a hydroxyl group on C3 only slightly reduces the affinity (Kd increased from 13.4 × 10−6 M for quercetin to 29.2 × 10−6 M for luteolin), while a covalently bound disaccharide at this position practically nullifies the binding. pH 2.5 significantly reduced such binding while the difference between 4.5 and 6.5 was small and structure dependent.

Physical and chemical characterization of the 2019 “black rain” event in the Metropolitan Area of São Paulo, Brazil

Aerosols emitted from biomass burning in South American tropical forests have been a concern in the last decades. On August 19th, 2019 darkened precipitation was observed over the metropolitan area of São Paulo (MASP), in an unprecedented event termed “black rain”, after intense biomass burning episodes in the South American hinterland. Satellite imagery, back trajectory analyses, and meteorological reports showed air masses transporting thick plumes of biomass burning aerosols originated in part from the Amazon Basin and Bolivia. Rainwater samples were collected in different sites of the metropolitan area, during and after the “black rain” event, and both physically and chemically characterized to assess the possible influence of biomass burning aerosols in the event. The collected samples in the “black rain” event presented high turbidity (above 70 NTU), and biomass burning organic tracers (levoglucosan, mannosan, and galactosan) were observed in higher concentrations in the “black rain” samples than in control ones (e.g. average levoglucosan of 0.33 μg mL−1, compared to 0.02 μg mL−1 after the event), with deposition fluxes more than three times higher during the event (1.04 mg m−2 and 0.31 mg m−2, respectively). The detection of glucose, xylose, and mannose, after hydrolysis of the solid material present in the “black rain” samples, strongly suggested the presence of plant cell wall material derived from the partial combustion of wood and grass matter. Total polycyclic aromatic hydrocarbons (PAHs) concentrations were, in general, higher during the event than in post-event control samples collected in the MASP (on average, almost 15 times higher than after the event), with a higher deposition flux (5.1 and 2.7 mg m−2, respectively). Overall, there are strong pieces of evidence that the long-range transport of smoke produced in South American forest fires was connected to the precipitation of darkened rainwater over the Metropolitan Area of São Paulo.

Genetic Dissection for Maize Forage Digestibility Traits in a Multi-Parent Advanced Generation Intercross (MAGIC) Population

Forage feedstock is the greatest source of energy for livestock. Unfortunately, less than 50% of their fiber content is actually digested and assimilated by the ruminant animals. This recalcitrance is mainly due to the high concentration of plant cell wall material and to the limited digestion of the fiber by the microorganisms. A Genome-Wide Association Study (GWAS) was carried out in order to identify Single Nucleotide Polymorphisms (SNPs) associated with forage digestibility traits in a maize Multi-Parent Advanced Generation Intercross (MAGIC) population. We identified seven SNPs, corresponding to five Quantitative Trait Loci (QTL), associated to digestibility of the organic matter, 11 SNPs, clustered in eight QTLs, associated to Neutral Detergent Fiber (NDF) content and eight SNPs corresponding with four QTL associated with Acid Detergent Fiber (ADF). Candidate genes under the QTL for digestibility of the organic matter could be the ones involved in pectin degradation or phenylpropanoid pathway. Transcription factor genes were also proposed for the fiber QTL identified, in addition to genes induced by oxidative stress, or a gene involved in lignin modifications. Nevertheless, for the improvement of the traits under study, and based on the moderate heritability value and low percentage of the phenotypic variability explained by each QTL, a genomic selection strategy using markers evenly distributed across the whole genome is proposed.

Unraveling the xylanolytic potential of Acidobacteria bacterium AB60 from Cerrado soils.

The presence of genes for glycosyl hydrolases in many Acidobacteria genomes indicates an important role in the degradation of plant cell wall material. Acidobacteria bacterium AB60 was obtained from Cerrado oligotrophic soil in Brazil, where this phylum is abundant. The 16S rRNA gene analyses showed that AB60 was closely related to the genera Occallatibacter and Telmatobacter. However, AB60 grew on xylan as carbon source, which was not observed in Occallatibacter species; but growth was not detected on medium containing carboxymethyl cellulose, as observed in Telmatobacter. Nevertheless, the genome analysis of AB60 revealed genes for the enzymes involved in cellulose as well as xylan degradation. In addition to enzymes involved in xylan degradation, α-l-rhamnosidase was detected in the cultures of AB60. Functional screening of a small-insert genomic library did not identify any clones capable of carboxymethyl cellulose degradation, but open reading frames coding α-l-arabinofuranosidase and α-l-rhamnosidase were present in clones showing xylan degradation halos. Both enzymes act on the lateral chains of heteropolymers such as pectin and some hemicelluloses. These results indicate that the hydrolysis of α-linked sugars may offer a metabolic niche for slow-growing Acidobacteria, allowing them to co-exist with other plant-degrading microbes that hydrolyze β-linked sugars from cellulose or hemicellulose backbones.

Mild Acetylation and Solubilization of Ground Whole Plant Cell Walls in EmimAc: A Method for Solution-State NMR in DMSO-d6.

Lignocellulosic biomass is mainly composed of polysaccharides and lignin. The complexity and diversity of the plant cell wall polymers makes it difficult to isolate the components in pure form for characterization. Many current approaches to analyzing the structure of lignocellulose, which involve sequential extraction and characterization of the resulting fractions, are time-consuming and labor-intensive. The present study describes a new and facile system for rationally derivatizing and dissolving coarsely ground plant cell wall materials. Using ionic liquids (EmimAc) and dichloroacetyl chloride as a solvent/reagent produced mildly acetylated whole cell walls without significant degradation. The acetylated products were soluble in DMSO-d6 from which they can be characterized by solution-state two-dimensional nuclear magnetic resonance (2D NMR) spectrometry. A distinct advantage of the procedure is that it realizes the dissolution of whole lignocellulosic materials without requiring harsh ball milling, thereby allowing the acquisition of high-resolution 2D NMR spectra to revealing structural details of the main components (lignin and polysaccharides). The method is therefore beneficial to understanding the composition and structure of biomass aimed at its improved utilization.

A catalog of microbial genes from the bovine rumen unveils a specialized and diverse biomass-degrading environment.

BackgroundThe rumen microbiota provides essential services to its host and, through its role in ruminant production, contributes to human nutrition and food security. A thorough knowledge of the genetic potential of rumen microbes will provide opportunities for improving the sustainability of ruminant production systems. The availability of gene reference catalogs from gut microbiomes has advanced the understanding of the role of the microbiota in health and disease in humans and other mammals. In this work, we established a catalog of reference prokaryote genes from the bovine rumen.ResultsUsing deep metagenome sequencing we identified 13,825,880 non-redundant prokaryote genes from the bovine rumen. Compared to human, pig, and mouse gut metagenome catalogs, the rumen is larger and richer in functions and microbial species associated with the degradation of plant cell wall material and production of methane. Genes encoding enzymes catalyzing the breakdown of plant polysaccharides showed a particularly high richness that is otherwise impossible to infer from available genomes or shallow metagenomics sequencing. The catalog expands the dataset of carbohydrate-degrading enzymes described in the rumen. Using an independent dataset from a group of 77 cattle fed 4 common dietary regimes, we found that only <0.1% of genes were shared by all animals, which contrast with a large overlap for functions, i.e., 63% for KEGG functions. Different diets induced differences in the relative abundance rather than the presence or absence of genes, which explains the great adaptability of cattle to rapidly adjust to dietary changes.ConclusionsThese data bring new insights into functions, carbohydrate-degrading enzymes, and microbes of the rumen to complement the available information on microbial genomes. The catalog is a significant biological resource enabling deeper understanding of phenotypes and biological processes and will be expanded as new data are made available.

Proteomic Characterization of Lignocellulolytic Enzymes Secreted by the Insect-Associated Fungus Daldinia decipiens oita, Isolated from a Forest in Northern Japan.

Wood-devastating insects utilize their symbiotic microbes with lignocellulose-degrading abilities to extract energy from recalcitrant woods. It is well known that free-living lignocellulose-degrading fungi secrete various carbohydrate-active enzymes (CAZymes) to degrade plant cell wall components, mainly cellulose, hemicellulose, and lignin. However, CAZymes from insect-symbiotic fungi have not been well documented except for a few examples. In this study, an insect-associated fungus, Daldinia decipiens oita, was isolated as a potential symbiotic fungus of female Xiphydria albopicta captured from Hokkaido forest. This fungus was grown in seven different media containing a single carbon source, glucose, cellulose, xylan, mannan, pectin, poplar, or larch, and the secreted proteins were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 128 CAZymes, including domains of 92 glycoside hydrolases, 15 carbohydrate esterases, 5 polysaccharide lyases, 17 auxiliary activities, and 11 carbohydrate-binding modules, were identified, and these are involved in degradation of cellulose and hemicellulose but not lignin. Together with the results of polysaccharide-degrading activity measurements, we concluded that D. decipiens oita tightly regulates the expression of these CAZymes in response to the tested plant cell wall materials. Overall, this study described the detailed proteomic approach of a woodwasp-associated fungus and revealed that the new isolate, D. decipiens oita, secretes diverse CAZymes to efficiently degrade lignocellulose in the symbiotic environment.IMPORTANCE Recent studies show the potential impacts of insect symbiont microbes on biofuel application with regard to their degradation capability of a recalcitrant plant cell wall. In this study, we describe a novel fungal isolate, D. decipiens oita, as a single symbiotic fungus from the Xiphydria woodwasp found in the northern forests of Japan. Our detailed secretome analyses of D. decipiens oita, together with activity measurements, reveal that this insect-associated fungus exhibits high and broad activities for plant cell wall material degradation, suggesting potential applications within the biomass conversion industry for plant mass degradation.

Nanostructure and poroviscoelasticity in cell wall materials from onion, carrot and apple: Roles of pectin

The hierarchical organisation of polysaccharides in primary plant cell walls is responsible for their unique mechanical properties, and in turn for the textural and rheological properties of plant-based foods and ingredients. It is expected that at the nano scale, the mechanical properties of cell wall materials arise from a combination of structural deformation of the polysaccharide networks and hydraulic properties of the continuous water phase, as has been shown for other cellulose-based composites. Pectin plays a key role in the load bearing properties of (bacterial) cellulose-pectin composites due to its contribution to both hydration structure and the dynamics of water movement. To investigate whether these features are also important in plant cell wall materials we have used a set of advanced characterisation techniques to elucidate cell wall structural features at different length scales (X-ray diffraction and small angle X-ray and neutron scattering) in cell walls from two dicotyledons (apple and carrot) and a non-commelinid monocotyledon (onion). The strength of isolated cell walls was measured under compression and fitted to a poroviscoelastic mechanical model, demonstrating that the mechanical properties of the isolated cell wall materials are directly linked to both polysaccharide networks and fluid flow through the networks. Our results show how pectin polysaccharides influence the viscoelastic behaviour of these materials and contribute to the texture of plant-derived food systems.

Biobased Cellulose Nanofibril-Oil Composite Films for Active Edible Barriers.

Low-concentration oil-in-water emulsions stabilized by cellulose nanofibrils (CNFs) extracted from primary plant cell wall materials are used to prepare thin biobased CNF–oil composite films by solvent casting. Flexible, transparent, and biodegradable composite films are obtained, with increased thermal stability (up to 300 °C) as the oil concentration increases. Examination of the microstructure demonstrates a clear dependency on the oil content, as a multilayered structure where the oil phase trapped between two layers of CNFs is appreciated at high oil concentrations. The embedded oil significantly influences the mechanical and wetting properties of the films, confirming their potential for use in packaging systems. Encapsulation of curcumin in the composite films leads to an increased antioxidant (up to 30% radical scavenging activity) and antimicrobial activity, inhibiting the growth of foodborne bacteria such as Escherichia coli. The resulting composite films show promising results in the field of active packaging for applications in the food, pharmaceutical, and cosmetic industries.

Drying of pickering emulsions in a viscoelastic network of cellulose microfibrils

Oil-in-water Pickering emulsions at high volume fractions (50 wt % oil) are prepared using cellulose microfibrils (CMF) from plant cell wall materials as emulsifiers and stabilizer. Confocal microscopy is used to monitor the changing CMF network over time and the rate of oil droplet coalescence. Without addition of glycerol the drying emulsion quickly coalesces, and a significant oil separation ensues. The rate of coalescence and the percentage of coalescence droplets and oil separation decrease as the concentration of CMF is increased. Addition of glycerol into the emulsions decreases the extent of droplet coalescence and oil separation. At 10 wt% added glycerol, coalescence could not be prevented but oil separation is minimal: the oil is mainly contained into honeycomb shaped cells made of CMF and glycerol. The results demonstrate the influence of the aqueous phase on the coalescence and film forming properties of CMF stabilized Pickering emulsions.

Root mediated uptake of Salmonella is different from phyto-pathogen and associated with the colonization of edible organs

BackgroundPre-harvest contamination of fruits and vegetables by Salmonella in fields is one of the causes of food-borne outbreaks. Natural openings like stomata, hydathodes and fruit cracks are known to serve as entry points. While there are reports indicating that Salmonella colonize and enter root through lateral root emerging area, further investigations regarding how the accessibility of Salmonella to lateral root is different from phyto-pathogenic bacteria, the efficacy of lateral root to facilitate entry have remained unexplored. In this study we attempted to investigate the lateral root mediated entry of Salmonella, and to bridge this gap in knowledge.ResultsUnlike phytopathogens, Salmonella cannot utilize cellulose as the sole carbon source. This negates the fact of active entry by degrading plant cellulose and pectin. Endophytic Salmonella colonization showed a high correlation with number of lateral roots. When given equal opportunity to colonize the plants with high or low lateral roots, Salmonella internalization was found higher in the plants with more lateral roots. However, the epiphytic colonization in both these plants remained unaltered. To understand the ecological significance, we induced lateral root production by increasing soil salinity which made the plants susceptible to Salmonella invasion and the plants showed higher Salmonella burden in the aerial organs.ConclusionSalmonella, being unable to degrade plant cell wall material relies heavily on natural openings. Therefore, its invasion is highly dependent on the number of lateral roots which provides an entry point because of the epidermis remodeling. Thus, when number of lateral root was enhanced by increasing the soil salinity, plants became susceptible to Salmonella invasion in roots and its transmission to aerial organs.

Unravelling the Mechanism of Stabilization and Microstructure of Oil-in-Water Emulsions by Native Cellulose Microfibrils in Primary Plant Cells Dispersions.

It is long known that oil-in-water emulsions can be stable against coalescence in homogenized plant cell wall dispersions because of the presence of surface-active biopolymers. When plant cell wall material is homogenized to the extent of deagglomeration of the cellulose microfibrils (CMFs), a much more complex dispersed system is obtained. Here we show that in such complex systems both surface active soluble polymers and individual CMFs are at the origin of this stabilization against coalescence, as they form a shell around the oil droplets providing Pickering-like stabilization. Individual CMFs and bundles of them in the presence of soluble biopolymers form a hybrid network in the continuous phase linking the droplets, creating a viscoelastic network that prevents the droplets from coalescing. Depletion induced attraction caused by soluble biopolymers and dispersed CMFs induces the formation of oil droplet clusters at low CMF concentrations leading to a highly heterogeneous distribution of oil droplets. This effect diminishes at high CMF concentrations at which the strong viscoelastic network arrests the droplets. These findings are important steps toward controlling complex dispersed systems comprising CMF-polymers mixtures with a second liquid or solid dispersed phase.

Effect of subcritical water and steam explosion pretreatments on the recovery of sterols, phenols and oil from olive pomace

Hydrothermal pretreatments including steam explosion and subcritical water (SCW) facilitate hydrolyses of plant cell wall materials and supply environmentally friendly extraction solvents to recover value added compounds. In this study, effect of steam and SCW and temperature (160, 180 and 200 °C, 5 min) on yield and β-sitosterol content of pomace oil and phenolics were compared for value added utilization of olive pomace. Compared to acid hydrolysis, hydrothermal pretreatments yielded similar oil recovery. 54–76% of the bound oil and 18–32% of the bound β-sitosterol of the pomace were recovered by hydrothermal pretreatments. Steam pretreatment was more effective than SCW pretreatment at lower temperatures, however SCW showed increasing trend on oil yield related to steam pretreatment. As further research on process development, use of sequential temperature might be investigated, starting with steam explosion (<180 °C), followed by SCW pretreatment (>200 °C) to obtain multiple, aqueous and meal, fractions for total valorization of olive pomace.