ABSTRACT
Cholinesterases, involved in acetylcholine catabolism in the central and peripheral nervous system, have been strongly linked with neurodegenerative diseases. Current therapeutic approaches using synthetic drugs present several side effects. Hence, there is an increasing research interest in naturally-occurring dietary polyphenols, which are also considered efficacious. Food processing by-products such as brewer's spent grain (BSG) would be a potential bio-source of polyphenols. In this study, polyphenol-rich BSG extracts using 60% acetone and 0.75% NaOH solutions were generated, which were further subjected to liquid-liquid partitioning using various organic solvents. The water-partitioned fractions of the saponified extracts had the highest total polyphenol content (6.2 ± 2.8 mgGAE/g dw) as determined by Folin-Ciocalteu reagent, while the LC-MS/MS showed ethyl acetate fraction with the highest phenolics (2.9 ± 0.3 mg/g BSG dw). The best inhibitions of acetyl- (37.9 ± 2.9%) and butyryl- (53.6 ± 7.7%) cholinesterases were shown by the diethyl ether fraction of the saponified extract. This fraction contained the highest sum of quantified phenolics (99 ± 21.2 µg/mg of extract), and with significant (p < 0.01) inhibitory contribution of decarboxylated-diferulic acid. Amongst the standards, caffeic acid presented the highest inhibition for both cholinesterases, 25.5 ± 0.2% for acetyl- and 52.3 ± 0.8% for butyryl-cholinesterase, respectively, whilst the blends insignificantly inhibited both cholinesterases. The results showed that polyphenol-rich BSG fractions have potentials as natural anti-cholinesterase agents.
ABSTRACT
The recovery of antioxidant polyphenols from light, dark and mix brewer's spent grain (BSG) using conventional maceration, microwave and ultrasound assisted extraction was investigated. Total polyphenols were measured in the crude (60% acetone), liquor extracts (saponified with 0.75% NaOH) and in their acidified ethyl acetate (EtOAc) partitioned fractions both by spectrophotometry involving Folin-Ciocalteu reagent and liquid-chromatography-tandem mass spectrometry (LC-MS/MS) methods. Irrespective of the extraction methods used, saponification of BSG yielded higher polyphenols than in the crude extracts. The EtOAc fractionations yielded the highest total phenolic content (TPC) ranging from 3.01 ± 0.19 to 4.71 ± 0.28 mg gallic acid equivalent per g of BSG dry weight. The corresponding total polyphenols quantified by LC-MS/MS ranged from 549.9 ± 41.5 to 2741.1 ± 5.2 µg/g of BSG dry weight. Microwave and ultrasound with the parameters and equipment used did not improve the total polyphenol yield when compared to the conventional maceration method. Furthermore, the spectrophotometric quantification of the liquors overestimated the TPC, while the LC-MS/MS quantification gave a closer representation of the total polyphenols in all the extracts. The total polyphenols were in the following order in the EtOAc fractions: BSG light > BSG Mix > BSG dark, and thus suggested BSG light as a sustainable, low cost source of natural antioxidants that may be tapped for applications in food and phytopharmaceutical industries.
ABSTRACT
BACKGROUND: Rice husk and rice straw represent promising sources of biomass for production of renewable fuels and chemicals. For efficient utilisation, lignocellulosic components must first be pretreated to enable efficient enzymatic saccharification and subsequent fermentation. Existing pretreatments create breakdown products such as sugar-derived furans, and lignin-derived phenolics that inhibit enzymes and fermenting organisms. Alkali pretreatments have also been shown to release significant levels of simple, free phenolics such as ferulic acid that are normally esterified to cell wall polysaccharides in the intact plant. These phenolics have recently been found to have considerable inhibitory properties. The aim of this research has been to establish the extent to which such free phenolic acids are also released during hydrothermal pretreatment of rice straw (RS) and rice husk (RH). RESULTS: RS and RH were subjected to hydrothermal pretreatments over a wide range of severities (1.57-5.45). FTIR analysis showed that the pretreatments hydrolysed and solubilised hemicellulosic moieties, leading to an enrichment of lignin and crystalline cellulose in the insoluble residue. The residues also lost the capacity for UV autofluorescence at pH 7 or pH 10, indicating the breakdown or release of cell wall phenolics. Saponification of raw RS and RH enabled identification and quantification of substantial levels of simple phenolics including ferulic acid (tFA), coumaric acid (pCA) and several diferulic acids (DiFAs) including 8-O-4'-DiFA, 8,5'-DiFA and 5,5'-DiFA. RH had higher levels of pCA and lower levels of tFA and DiFAs compared with RS. Assessment of the pretreatment liquors revealed that pretreatment-liberated phenolics present were not free but remained as phenolic esters (at mM concentrations) that could be readily freed by saponification. Many were lost, presumably through degradation, at the higher severities. CONCLUSION: Differences in lignin, tFA, DiFAs and pCA between RS and RH reflect differences in cell wall physiology, and probably contribute to the higher recalcitrance of RH compared with RS. Hydrothermal pretreatments, unlike alkali pretreatments, release cinnamic acid components as esters. The potential for pretreatment-liberated phenolic esters to be inhibitory to fermenting microorganisms is not known. However, the present study shows that they are found at concentrations that could be significantly inhibitory if released as free forms by enzyme activity.
ABSTRACT
Approximately 88 Million tonnes (Mt) of food is wasted in the European Union each year and the environmental impacts of these losses throughout the food supply chain are widely recognised. This study illustrates the impacts of food waste in relation to the total food utilised, including the impact from food waste management based on available data at the European level. The impacts are calculated for the Global Warming Potential, the Acidification Potential and the Eutrophication Potential using a bottom-up approach using more than 134 existing LCA studies on nine representative products (apple, tomato, potato, bread, milk, beef, pork, chicken, white fish). Results show that 186 Mt CO2-eq, 1.7 Mt SO2-eq. and 0.7 Mt PO4-eq can be attributed to food waste in Europe. This is 15 to 16% of the total impact of the entire food supply chain. In general, the study confirmed that most of the environmental impacts are derived from the primary production step of the chain. That is why animal-containing food shows most of the food waste related impacts when it is extrapolated to total food waste even if cereals are higher in mass. Nearly three quarters of all food waste-related impacts for Global Warming originate from greenhouse gas emissions during the production step. Emissions by food processing activities contribute 6%, retail and distribution 7%, food consumption, 8% and food disposal, 6% to food waste related impacts. Even though the results are subject to certain data and scenario uncertainties, the study serves as a baseline assessment, based on current food waste data, and can be expanded as more knowledge on the type and amount of food waste becomes available. Nevertheless, the importance of food waste prevention is underlined by the results of this study, as most of the impacts originate from the production step. Through food waste prevention, those impacts can be avoided as less food needs to be produced.
Subject(s)
Environment , Food , Global Warming , Waste Management , Animals , EuropeABSTRACT
BACKGROUND: Rice straw and husk are globally significant sources of cellulose-rich biomass and there is great interest in converting them to bioethanol. However, rice husk is reportedly much more recalcitrant than rice straw and produces larger quantities of fermentation inhibitors. The aim of this study was to explore the underlying differences between rice straw and rice husk with reference to the composition of the pre-treatment liquors and their impacts on saccharification and fermentation. This has been carried out by developing quantitative NMR screening methods. RESULTS: Air-dried rice husk and rice straw from the same cultivar were used as substrates. Carbohydrate compositions were similar, whereas lignin contents differed significantly (husk: 35.3% w/w of raw material; straw 22.1% w/w of raw material). Substrates were hydrothermally pre-treated with high-pressure microwave processing across a wide range of severities. 25 compounds were identified from the liquors of both pre-treated rice husk and rice straw. However, the quantities of compounds differed between the two substrates. Fermentation inhibitors such as 5-HMF and 2-FA were highest in husk liquors, and formic acid was higher in straw liquors. At a pre-treatment severity of 3.65, twice as much ethanol was produced from rice straw (14.22% dry weight of substrate) compared with the yield from rice husk (7.55% dry weight of substrate). Above severities of 5, fermentation was inhibited in both straw and husk. In addition to inhibitors, high levels of cellulase-inhibiting xylo-oligomers and xylose were found and at much higher concentrations in rice husk liquor. At low severities, organic acids and related intracellular metabolites were released into the liquor. CONCLUSIONS: Rice husk recalcitrance to saccharification is probably due to the much higher levels of lignin and, from other studies, likely high levels of silica. Therefore, if highly polluting chemical pre-treatments and multi-step biorefining processes are to be avoided, rice husk may need to be improved through selective breeding strategies, although more careful control of pre-treatment may be sufficient to reduce the levels of fermentation inhibitors, e.g. through steam explosion-induced volatilisation. For rice straw, pre-treating at severities of between 3.65 and 4.25 would give a glucose yield of between 37.5 and 40% (w/DW, dry weight of the substrate) close to the theoretical yield of 44.1% w/DW, and an insignificant yield of total inhibitors.
ABSTRACT
Lodging continues to be a major cause of yield loss in important crop species such as Brassica napus. Understanding the genetic regulation of lodging resistance is therefore of key interest to breeders worldwide. Current strategies aimed at minimising lodging risk involve the incorporation of dwarfing genes or the application of plant growth regulators. However, despite these efforts, lodging continues to be a persistent problem and it is therefore of high interest that novel, complimentary strategies for lodging control are implemented. One approach would be to focus on understanding the genetic properties underlying stem mechanical strength. With this in mind, we screened a training genetic diversity panel of B. napus accession for variation in stem mechanical strength and related traits. Using Associative Transcriptomics, we identified molecular markers for a suite of valuable traits. Using an independent test genetic diversity panel, we show that the methods employed are robust for identification of predictive markers. Furthermore, based on conserved synteny with Arabidopsis thaliana, we are able to provide a biological context to the marker associations detected and provide evidence for a role in pectin methylesterification in contributing to stem mechanical strength in Brassicaceae.
ABSTRACT
PURPOSE: The term bioaccessibility refers to the proportion of a nutrient released from a complex food matrix during digestion and, therefore, becoming potentially available for absorption in the gastrointestinal tract. In the present study, we assessed the starch and protein bioaccessibility from a range of wheat endosperm products differing in particle size. METHODS: Five porridge meals (size A, flour, mean particle size 0.11 mm, size B, small, mean particle size 0.38 mm, size C, semolina, mean particle size 1.01 mm, size D, medium, mean particle size 1.44 mm, size E, large, mean particle size 1.95 mm) with theoretically different postprandial glycaemic responses were subjected to oral processing in vitro, followed by simulated gastric and duodenal digestion. RESULTS: A significant increase (P < 0.001) in starch degradation was observed in size A (52%) compared with size E (25%). Both sizes C and D gave less, although not significantly, digestible starch (32 and 28%, respectively). The glucose release significantly decreased as the particle size of the meal increased (92.16% detected for size A vs 47.39% for size E). In agreement with starch degradation and glucose release, size A gave the most digestible protein. CONCLUSIONS: This data provide further evidence that, by decreasing the size of wheat endosperm, starch release and glycaemic response are enhanced. We also showed that protein bioaccessibility followed a similar trend as for starch digestion. Finally, these results support the hypothesis that different degrees of starch encapsulation elicit different blood glucose responses.
Subject(s)
Digestion , Edible Grain/chemistry , Particle Size , Plant Proteins/metabolism , Starch/metabolism , Triticum , Amylases/metabolism , Bile/metabolism , Biological Availability , Blood Glucose/metabolism , Duodenum/metabolism , Gastric Mucosa/enzymology , Glucose/metabolism , Humans , Lipase/metabolism , Pancreas/enzymology , Pepsin A/metabolism , Saliva/immunology , Starch/pharmacokineticsABSTRACT
BACKGROUND: Wheat straw forms an important, reliable source of lignocellulosic biomass for use in second-generation ethanol production. However, there is limited understanding of the variation in quality of straw from current breeding cultivars, and studies on such variation have generally employed suboptimal pretreatments. There is also a degree of confusion regarding phenotypic characteristics relevant to optimising the enzymatic saccharification of cellulose after suitable pretreatments for biorefining compared with those which determine good ruminant digestibility. The aim of this study has been to (a) evaluate and compare the levels of glucose enzymatically released from straw obtained from 89 cultivars of winter wheat after optimised hydrothermal pretreatments and (b) identify the underlying phenotypic characteristics relevant to enhanced glucose production with special reference to the ratios of constituent tissue types. RESULTS: Optimised pretreatment involved hydrothermal extraction at 210 °C for 10 min. Using excess cellulases, quantitative saccharification was achieved within 24 h. The amount of glucose released ranged from 192 to 275 mg/g. The extent of glucose release was correlated with (a) the level of internode tissue (R = 0.498; p = 6.84 × 10-7), (b) stem height (R = 0.491; p = 1.03 × 10-6), and (c) chemical characteristics particular to stem tissues including higher levels of cellulose (R = 0.552; p = 2.06 × 10-8) and higher levels of lignin R = 0.494; p = 8.67 × 10-7. CONCLUSIONS: In order to achieve maximum yields of cellulosic glucose for second-generation ethanol production, a predisposition for wheat to produce cellulose-enriched internode stem tissue, particularly of longer length, would be beneficial. This contrasts with the ideotype for ruminant nutrition, in which an increased proportion of leaf tissue is preferable.
ABSTRACT
This study compares in vitro and in vivo models of lipid digestion from almond particles within a complex food matrix (muffins) investigating whether the cell-wall barrier regulates the bioaccessibility of nutrients within this matrix. Muffins containing small (AF) or large (AP) particles of almond were digested in triplicate using an in vitro dynamic gastric model (DGM, 1 h) followed by a static duodenal digestion (8 h). AF muffins had 97.1 ± 1.7% of their lipid digested, whereas AP muffins had 57.6 ± 1.1% digested. In vivo digestion of these muffins by an ileostomy volunteer (0-10 h) gave similar results with 96.5% and 56.5% lipid digested, respectively. The AF muffins produced a higher postprandial triacylglycerol iAUC response (by 61%) than the AP muffins. Microstructural analysis showed that some lipid remained encapsulated within the plant tissue throughout digestion. The cell-wall barrier mechanism is the main factor in regulating lipid bioaccessibility from almond particles.
ABSTRACT
In addition to ethanol, yeasts have the potential to produce many other industrially-relevant chemicals from numerous different carbon sources. However there remains a paucity of information about overall capability across the yeast family tree. Here, 11 diverse species of yeasts with genetic backgrounds representative of different branches of the family tree were investigated. They were compared for their abilities to grow on a range of sugar carbon sources, to produce potential platform chemicals from such substrates and to ferment hydrothermally pretreated rice straw under simultaneous saccharification and fermentation conditions. The yeasts differed considerably in their metabolic capabilities and production of ethanol. A number could produce significant amounts of ethyl acetate, arabinitol, glycerol and acetate in addition to ethanol, including from hitherto unreported carbon sources. They also demonstrated widely differing efficiencies in the fermentation of sugars derived from pre-treated rice straw biomass and differential sensitivities to fermentation inhibitors. A new catabolic property of Rhodotorula mucilaginosa (NCYC 65) was discovered in which sugar substrate is cleaved but the products are not metabolised. We propose that engineering this and some of the other properties discovered in this study and transferring such properties to conventional industrial yeast strains could greatly expand their biotechnological utility.
Subject(s)
Biodiversity , Biofuels/microbiology , Rhodotorula/metabolism , Carbon/metabolism , Fermentation , Oryza/chemistry , Rhodotorula/growth & development , Species Specificity , Sugars/metabolismABSTRACT
BACKGROUND: In Uganda, the chaff remaining from threshed panicles of millet and sorghum is a low value, lignocellulose-rich agricultural by-product. Currently, it is used as a substrate for the cultivation of edible Oyster mushrooms (Pleurotus ostreatus). The aim of this study was to assess the potential to exploit the residual post-harvest compost for saccharification and fermentation to produce ethanol. RESULTS: Sorghum and millet chaff-derived spent oyster mushroom composts minus large mycelium particles were assessed at small-scale and low substrate concentrations (5% w/v) for optimal severity hydrothermal pre-treatment, enzyme loading and fermentation with robust yeasts to produce ethanol. These conditions were then used as a basis for larger scale assessments with high substrate concentrations (30% w/v). Millet-based compost had a low cellulose content and, at a high substrate concentration, did not liquefy effectively. The ethanol yield was 63.9 g/kg dry matter (DM) of original material with a low concentration (19.6 g/L). Compost derived from sorghum chaff had a higher cellulose content and could be liquefied at high substrate concentration (30% w/v). This enabled selected furfural-resistant yeasts to produce ethanol at up to 186.9 g/kg DM of original material and a concentration of 45.8 g/L. CONCLUSIONS: Spent mushroom compost derived from sorghum chaff has the potential to be an industrially useful substrate for producing second-generation bioethanol. This might be improved further through fractionation and exploitation of hemicellulosic moieties, and possibly the exploitation of the mycelium-containing final residue for animal feed. However, spent compost derived from millet does not provide a suitably high concentration of ethanol to make it industrially attractive. Further research on the difficulty in quantitatively saccharifying cellulose from composted millet chaff and other similar substrates such as rice husk is required.
ABSTRACT
Plant cell wall materials derived from a range of waste biomass sources have great potential as a source of sustainable alternatives to petrochemicals. Perhaps the most straightforward way of realising this potential would be to hydrolyse the most efficiently fermentable polymers into their constituent sugars and use yeast to ferment these into useful chemicals. However, it also makes sense to pre-extract components which have a greater value in polymeric form. This is particularly true for non-cellulosic polymers, which are rich in poorly-fermentable pentose sugars. Liquid hot water (LHW) pretreatment can be used to extract non-cellulosic carbohydrates in a cost-effective manner, leaving a cellulose-rich substrate which is easier to hydrolyse using commercial cellulases. However, inherent differences in the plant cell wall structure and composition mean that some biomass sources may be more suitable for exploitation than others. Here, we examine eight different feedstocks (two each from hardwood, softwood, cereal straws and dicotyledonous crops), expose them to 26 different LHW pretreatment conditions and hydrolyse the entire pretreated slurry with a commercial cellulase. This enables side-by-side comparisons, in terms of saccharification yield, of the feedstocks. The results clearly demonstrate considerable differences in suitability between the feedstocks, in relation to the quantity of products released and the processes needed to obtain them.
Subject(s)
Polymers/chemistry , Microwaves , TemperatureABSTRACT
Genetic improvement of the plant cell wall has enormous potential to increase the quality of food, fibers, and fuels. However, the identification and characterization of genes involved in plant cell wall synthesis is far from complete. Association mapping is one of the few techniques that can help identify candidate genes without relying on our currently incomplete knowledge of cell wall synthesis. However, few cell wall phenotyping methodologies have proven sufficiently precise, robust, or scalable for association mapping to be conducted for specific cell wall polymers. Here, we created high-density carbohydrate microarrays containing chemically extracted cell wall polysaccharides collected from 331 genetically diverse Brassica napus cultivars and used them to obtain detailed, quantitative information describing the relative abundance of selected noncellulosic polysaccharide linkages and primary structures. We undertook genome-wide association analysis of data collected from 57 carbohydrate microarrays and identified molecular markers reflecting a diversity of specific xylan, xyloglucan, pectin, and arabinogalactan moieties. These datasets provide a detailed insight into the natural variations in cell wall carbohydrate moieties between B. napus genotypes and identify associated markers that could be exploited by marker-assisted breeding. The identified markers also have value beyond B. napus for functional genomics, facilitated by the close genetic relatedness to the model plant Arabidopsis Together, our findings provide a unique dissection of the genetic architecture that underpins plant cell wall biosynthesis and restructuring.
Subject(s)
Brassica napus/metabolism , Carbohydrate Metabolism/physiology , Carbohydrates , Cell Wall/metabolism , Databases, Factual , Microarray Analysis , Species SpecificityABSTRACT
Pectin and cellulose nanocrystals (CNCs) isolated from banana peels were used to prepare films. The effects of a reinforcing phase (CNCs) and a crosslinker (citric acid, CA) on properties of pectin films were studied. Glycerol-plasticized films were prepared by casting, with different CNC contents (0-10wt%), with or without CA. Overall tensile properties were improved by intermediate CNC contents (around 5wt%). The water resistance and water vapor barrier properties were also enhanced by CNC. Evidences were found from Fourier Transform Infrared (FTIR) spectra supporting the occurrence of crosslinking by CA. Additionally, the tensile strength, water resistance and barrier to water vapor were improved by the presence of CA. The 13C ssNMR spectra indicated that both CA and CNC promoted stiffening of the polymer chains.
Subject(s)
Musa/chemistry , Nanocomposites/chemistry , Pectins/chemistry , Cellulose/chemistry , Citric Acid/chemistry , Nanoparticles/chemistryABSTRACT
Wheat straw has been used as a source of hemicelluloses (WSH) and cellulose nanocrystals (CNC) for the elaboration of biodegradable films. Different films have been formed by using WSH as a matrix and different contents of CNC and citric acid. The predominant hemicelluloses were arabinoxylans. CNC reinforced the films, improving tensile strength and modulus, water resistance and water vapor barrier. Citric acid, on the other hand, presented concomitant plasticizing and crosslinking effects (the latter also evidenced by FTIR), probably due to a crosslinking extension by glycerol. The use of 5.9wt% CNC and 30wt% citric acid was defined as optimal conditions, resulting in minimum water sensitivity and permeability, while maintaining a good combination of tensile properties. Under those conditions, the films presented enhanced modulus, elongation, water resistance, and barrier to water vapor when compared to the control WSH film, and might be used for wrapping or coating a variety of foods.
ABSTRACT
BACKGROUND: Rice cultivation produces two waste streams, straw and husk, which could be exploited more effectively. Chemical pretreatment studies using rice residues have largely focussed on straw exploitation alone, and often at low substrate concentrations. Moreover, it is currently not known how rice husk, the more recalcitrant residue, responds to steam explosion without the addition of chemicals. RESULTS: The aim of this study has been to systematically compare the effects of steam explosion severity on the enzymatic saccharification and simultaneous saccharification and fermentation of rice straw and husk produced from a variety widely grown in Vietnam (Oryza sativa, cv. KhangDan18). Rice straw and husk were steam exploded (180-230 °C for 10 min) into hot water and washed to remove fermentation inhibitors. In both cases, pretreatment at 210 °C and above removed most of the noncellulosic sugars. Prolonged saccharification at high cellulase doses showed that rice straw could be saccharified most effectively after steam explosion at 210 °C for 10 min. In contrast, rice husk required more severe pretreatment conditions (220 °C for 10 min), and achieved a much lower yield (75 %), even at optimal conditions. Rice husk also required a higher cellulase dose for optimal saccharification (10 instead of 6 FPU/g DM). Hemicellulase addition failed to improve saccharification. Small pilot scale saccharification at 20 % (w/v) substrate loading in a 10 L high torque bioreactor resulted in similarly high glucose yields for straw (reaching 9 % w/v), but much less for husk. Simultaneous saccharification and fermentation under optimal pretreatment and saccharification conditions showed similar trends, but the ethanol yield from the rice husk was less than 40 % of the theoretical yield. CONCLUSIONS: Despite having similar carbohydrate compositions, pretreated rice husk is much less amenable to saccharification than pretreated rice straw. This is likely to attenuate its use as a biorefinery feedstock unless improvements can be made either in the feedstock through breeding and/or modern biotechnology, or in the pretreatment through the employment of improved or alternative technologies. Physiological differences in the overall chemistry or structure may provide clues to the nature of lignocellulosic recalcitrance.