Aiming for the complete utilization of sugar-beet pulp: Examination of the effects of mild acid and hydrothermal pretreatment followed by enzymatic digestion

<p>Abstract</p> <p>Background</p> <p>Biomass use for the production of bioethanol or platform chemicals requires efficient breakdown of biomass to fermentable monosaccharides. Lignocellulosic feedstocks often require physicochemical pretreatment before enzymatic hydrolysis can begin. The optimal pretreatment can be different for different feedstocks, and should not lead to biomass destruction or formation of toxic products.</p> <p>Methods</p> <p>We examined the influence of six mild sulfuric acid or water pretreatments at different temperatures on the enzymatic degradability of sugar-beet pulp (SBP).</p> <p>Results</p> <p>We found that optimal pretreatment at 140°C of 15 minutes in water was able to solubilize 60% w/w of the total carbohydrates present, mainly pectins. More severe treatments led to the destruction of the solubilized sugars, and the subsequent production of the sugar-degradation products furfural, hydroxymethylfurfural, acetic acid and formic acid. The pretreated samples were successfully degraded enzymatically with an experimental cellulase preparation.</p> <p>Conclusions</p> <p>In this study, we found that pretreatment of SBP greatly facilitated the subsequent enzymatic degradation within economically feasible time ranges and enzyme levels. In addition, pretreatment of SBP can be useful to fractionate functional ingredients such as arabinans and pectins from cellulose. We found that the optimal combined severity factor to enhance the enzymatic degradation of SBP was between log R'₀= -2.0 and log R'₀= -1.5. The optimal pretreatment and enzyme treatment solubilized up to 80% of all sugars present in the SBP, including ≥90% of the cellulose.</p

The influence of calcium on pectin's impact on TLR2 signalling

High intake of dietary fibres and calcium has been correlated to a lower frequency of Western disease such as allergy, asthma and obesity. How the combined higher intake of dietary fibres and calcium reduces the incidence of these diseases is unknown. Dietary fibre pectin can interact with Toll-like receptor (TLR) 2 and calcium in a degree of methyl-esterification (DM)-dependent manner. Low DM pectins interact stronger with TLR2 than high DM pectins. Since low DM pectin are known to bind calcium strongly, we investigated how calcium influences the DM-dependent impact of pectins on TLR2 signalling. We tested TLR2 activating, inhibiting and binding properties of pectins with DM18, DM52 and DM69 under 0 mM, 1 mM and 10 mM calcium conditions. None of the pectins activated TLR2, but pectins inhibited TLR2. Under 0 mM calcium conditions, especially DM18 and DM52 strongly inhibited TLR2 and bound strongly to TLR2. Addition of 1 and 10 mM calcium to these pectins reduced TLR2 inhibition and TLR2 binding. Our study shows that calcium reduces inhibition of TLR2 by low and intermediate DM pectins, but calcium has lower impact on TLR2 inhibition by high DM pectins. Calcium may therefore beneficially influence the impact of pectin on TLR2 signalling and contribute to an improved intestinal barrier function. A combined higher intake of pectin and calcium may therefore contribute to a lower incidence of Western diseases.</p

Impact of xylanase and glucanase on oligosaccharide formation, carbohydrate fermentation patterns, and nutrient utilization in the gastrointestinal tract of broilers

This study aimed at determining how the degradation of cereal non-starch polysaccharides (NSP) by dietary enzymes during feed digestion can influence nutrient digestibility and NSP fermentability in broilers. Ninety-six one-day-old male broilers were assigned to 4 different treatments: control and enzyme-supplemented wheat-based (WC, WE) or maize-based (MC, ME) treatments. Enzyme supplementation with endo-xylanase and endo-glucanase occurred from day 20 onwards. On day 28, digesta samples were collected. Nutrient digestibility, NSP recovery, oligosaccharide profile, and short-chain fatty acids (SCFA) content were determined. Enzyme supplementation in WE resulted in a higher starch (3%; p = 0.004) and protein (5%; p = 0.002) digestion in the ileum compared to WC. Xylanase activity in WE led to in situ formations of arabinoxylan-oligosaccharides consisting of 5 to 26 pentose units in the ileum. This coincided with decreased arabinose (p = 0.059) and xylose (p = 0.036) amounts in the ceca and higher acetate (p = 0.014) and butyrate (p = 0.044) formation in WE compared to WC. Conversely, complete total tract recovery of arabinoxylan in MC and ME suggested poor maize NSP fermentability. Overall, enzyme action improved nutrient digestibility and arabinoxylan fermentability in the wheat-based diet. The lower response of the maize-based diet to enzyme treatment may be related to the recalcitrance of maize arabinoxylan as well as to the high nutritive value of maize

Revealing methyl-esterification patterns of pectins by enzymatic fingerprinting:Beyond the degree of blockiness

Citrus pectins were studied by enzymatic fingerprinting using a simultaneous enzyme treatment with endo-polygalacturonase (endo-PG) from Kluyveromyces fragilis and pectin lyase (PL) from Aspergillus niger to reveal the methyl-ester distribution patterns over the pectin backbone. Using HILIC-MS combined with HPAEC enabled the separation and identification of the diagnostic oligomers released. Structural information on the pectins was provided by using novel descriptive parameters such as degree of blockiness of methyl-esterified oligomers by PG (DBPGme) and degree of blockiness of methyl-esterified oligomers by PL (DBPLme). This approach enabled us to clearly differentiate citrus pectins with various methyl-esterification patterns. The simultaneous use of PG and PL showed additional information, which is not revealed in digests using PG or PL alone. This approach can be valuable to differentiate pectins having the same DM and to get specific structural information on pectins and therefore to be able to better predict their physical and biochemical functionalities

Digestion, fermentation, and pathogen anti-adhesive properties of the hMO-mimic di-fucosyl-ß-cyclodextrin

Human milk is widely acknowledged as the best food for infants, and that is not just because of nutritional features. Human milk also contains a plethora of bioactive molecules, including a large set of human milk oligosaccharides (hMOs). Especially fucosylated hMOs have received attention for their anti-adhesive effects on pathogens, preventing attachment to the intestine and infection. Because hMOs are generally challenging to produce in sufficient quantities to study and ultimately apply in (medical) infant formula, novel compounds that are inspired by hMO structures (so-called "mimics") are interesting compounds to produce and evaluate for their biological effects. Here we present our thorough study into the digestion, fermentation and anti-adhesive capacity of the novel compound di-fucosyl-ß-cyclodextrin (DFßCD), which was inspired by the molecular structures of hMOs. We establish that DFßCD is not digested by a-amylase and also resistant to fermentation by microbial enzymes from a 9 month-old infant inoculum. In addition, we reveal that DFßCD blocks adhesion of enterotoxigenic E. coli (ETEC) to Caco-2 cells, especially when DFßCD is pre-incubated with ETEC prior to addition to the Caco-2 cells. This suggests that DFßCD functions through a decoy effect. We expect that our results inspire the generation and biological evaluation of other fucosylated hMOs and mimics, to obtain a comprehensive overview of the anti-adhesive power of fucosylated glycans. This journal i

The impact of the level and distribution of methyl-esters of pectins on TLR2-1 dependent anti-inflammatory responses

Pectins have anti-inflammatory effects via Toll-like receptor (TLR) inhibition in a degree of methyl-esterification-(DM)-dependent manner. However, pectins also vary in distribution of methyl-esters over the galactumnic-acid (GalA) backbone (Degree of Blockiness - DB) and impact of this on anti-inflammatory capacity is unknown. Pectins mainly inhibit TLR2-1 but magnitude depends on both DM and DB. Low DM pectins (DM18/19) with both low (DB86) and high DB (DB94) strongly inhibit TLR2-1. However, pectins with intermediate DM (DM43/ DM49) and high DB (DB60), but not with low DB (DB33), inhibit TLR2-1 as strongly as low DM. High DM pectins (DM84/88) with DB71 and DB91 do not inhibit TLR2-1 strongly. Pectin-binding to TLR2 was confirmed by capture-ELISA. In human macrophages, low DM and intermediate DM pectins with high DB inhibited TLR2-1 induced IL-6 secretion. Both high number and blockwise distribution of non-esterified GalA in pectins are responsible for the anti-inflammatory effects via inhibition of TLR2-1

Самосборка низкоразмерных систем Al, Ni, Ti и Si в условиях околоравновесной конденсации

Дисертація присвячена вивченню закономірностей структуро- утворення тривимірних мікро- та наносистем Al, Ni, Ti та Si за умов близькорівноважної стаціонарної конденсації. Вивчена залежність механізмів структуроутворення конденсату від основних параметрів технологічного процесу: часу конденсації, тиску робочого газу та потужності розряду. Проаналізовані фізичні процеси, що відбуваються під час роботи накопичувальних систем плазма-конденсат. Для цих систем розроблено математичну модель самоорганізації критично малих стаціонарних пересичень, яка вивчена за допомогою фазової площини. Досліджено вплив на кінетику формування мікро- та наносистем Al, Ni, Ti та Si взаємозв’язаних ефекту Гіббса-Томсона та польової селективності. Розроблені та апробовані за допомогою побудови фазових траєкторій математичні моделі, які описують процес самозбірки низькорозмірних систем. Встановлено, що за близько-рівноважних умов можна реалізувати різні структурно-морфологічні форми конденсатів. При цитуванні документа, використовуйте посилання http://essuir.sumdu.edu.ua/handle/123456789/24862Диссертация посвящена изучению закономерностей структуро- образования микро- и наносистем Al, Ni, Ti и Si в условиях критически малых пересыщений. Изучены процессы формирования конденсатов под влиянием структурной и полевой селективностей, степень проявления которых зависит от таких основных технологических параметров, как время конденсации, давление рабочего газа и мощность разряда. Проанализированы конструкции существующих распылитель- ных систем, разработана и создана накопительная система плазма- конденсат (НСПК) на базе планарного магнетрона и совмещенного с ним полого катода. Проанализированы происходящие в НСПК физические процессы и разработана математическая модель, описы- вающая изменение во времени таких нелинейно взаимозависимых технологических параметров, как пересыщение и температура ростовой поверхности. Методом фазовой плоскости установлена возможность самоорганизации критически малых стационарных пресыщений. С помощью математического моделирования показано, что при повышении давления рабочего газа происходит усреднение энергии распыленных атомов, что является необходимым условием повышения стационарности технологического процесса. Изучен механизм формирования фракталов Ni при осаждении предельно слабых паровых потоков на нагретые до относительно высоких температур (620 К) подложки. Показано, что снижение пересыщения паров Ni до критического значения может предельно локализировать области закрепления атомов и является основой для формирования на ростовой поверхности вискеров. Впервые показано, что диссипативная самоорганизация предельно малых стационарных пересыщений в накопительных системах плазма-конденсат приводит к консервативной самоорганизации низкоразмерных систем в виде тех или иных структурно-морфологических состояний поверхности конденсата. Изучены механизмы самосборки пористых пленок Al, трехмерных систем Ti в виде слабо связанных друг с другом ограненных или округлых кристаллов. Показано, что основой такой самосборки конденсатов является предельная минимизация свободной энергии конденсата. При этом селективная самосборка развитой поверхности металлов при проявлении полевой селективности является следствием выравнивания химических потенциалов в различных точках ростовой поверхности до уровня, при котором локальные скорости наращивания конденсатов не зависят от координат поверхности и не изменяются во времени. Изучены механизмы структурообразования конденсатов Si и разработана математическая модель, описывающая самоорганизацию островков одинаковых размеров и формы. При цитировании документа, используйте ссылку http://essuir.sumdu.edu.ua/handle/123456789/24862The aim of dissertation is the study of regularities of Al, Ni, Ti and Si micro- and nanosystems formation under the conditions of critically small supersaturations. It have been studied the processes of condensates forming at influence of structural and field selectivity, they are depend on main technological parameters of condensation process such as time of condensation, pressure of working gas and power discharge. Physical processes in the accumulative ion-plasma system have been analyzed. For this systems was created mathematical model of selforganization extremely low steady-state supersaturation and studied by method of phase plane. The influence of interdependent effects of Gibbs-Thomson and field selectivities on forming kinetic of micro- and nanosystems of Al, Ni, Ti and Si was investigated. It was created and tested due to plotting of phase trajectories mathematical models, which reply for the self-assembly process of low dimensional systems. It was established that different structuralmorphological forms of condensates could be obtain at near equilibrium conditions. When you are citing the document, use the following link http://essuir.sumdu.edu.ua/handle/123456789/2486

beta(2 -&gt; 1) chicory and beta(2 -&gt; 1)-beta(2 -&gt; 6) agave fructans protect the human intestinal barrier function in vitro in a stressor-dependent fashion

Dietary fibers such as fructans can protect the intestinal epithelial barrier integrity, but the mechanisms underlying this protection are not completely understood. We aimed to study the protective effect of β(2→1)-β(2→6) branched graminan-type fructans (GTFs) on gut epithelial barrier function that was disrupted by three different agents which impact the barrier function via different cellular mechanisms. The effects of GTFs were compared with those of linear β(2→1) inulin-type fructans (ITFs). T84 intestinal epithelial monolayers were incubated with GTFs and ITFs. Afterwards, the monolayers were challenged with the barrier disruptors calcium ionophore A23187, 12-myristate 13-acetate (PMA) and deoxynivalenol (DON). Transepithelial resistance was measured with an electric cell-substrate impedance sensing system. All fructans studied prevented the barrier disruption induced by A23187. ITF II protected from the disruptive effects of PMA. However, none of the studied fructans influenced the disruption induced by DON. As a measure of disruption-induced inflammation, interleukin-8 (IL-8) production by the intestinal epithelium was determined by ELISA. The production of IL-8 induced by A23187 was decreased by all fructans, whereas IL-8 production induced by DON decreased only upon pre-treatment with ITF II. None of the studied fructans prevented PMA induced IL-8 production. GTFs just like ITFs can influence the barrier function and inflammatory processes in gut epithelial cells in a structure-dependent fashion. These distinct protective effects are dependent on the different signaling pathways that lead to gut barrier disruption

Dietary Isomalto/Malto-Polysaccharides Increase Fecal Bulk and Microbial Fermentation in Mice

Scope: The prevalence of metabolic-syndrome-related disease has strongly increased. Nutritional intervention strategies appear attractive, particularly with novel prebiotics. Isomalto/malto-polysaccharides (IMMPs) represent promising novel prebiotics that promote proliferation of beneficial bacteria in vitro. The present study investigates for the first time the in vivo effects of IMMP in mice. Methods and results: C57BL/6 wild-type mice received control or IMMP-containing (10%, w/w) diets for 3 weeks. IMMP leads to significantly more fecal bulk (+26%, p < 0.05), higher plasma non-esterified fatty acids (colorimetric assay, +10%, p < 0.05), and lower fecal dihydrocholesterol excretion (mass spectrometry, −50%, p < 0.05). Plasma and hepatic lipid levels (colorimetric assays following lipid extraction) are not influenced by dietary IMMP, as are other parameters of sterol metabolism, including bile acids (gas chromatography/mass spectrometry). IMMP is mainly fermented in the cecum and large intestine (high-performance anion exchange chromatography). Next-generation sequencing demonstrates higher relative abundance of Bacteroides and butyrate producers (Lachnospiraceae, Roseburia Odoribacter) in the IMMP group. Conclusion: The combined results demonstrate that IMMP administration to mice increases fecal bulk and induces potentially beneficial changes in the intestinal microbiota. Further studies are required in disease models to substantiate potential health benefits.</p

Distinct fermentation of human milk oligosaccharides 3-FL and LNT2 and GOS/inulin by infant gut microbiota and impact on adhesion of Lactobacillus plantarum WCFS1 to gut epithelial cells

Human milk oligosaccharides (hMOs) are unique bioactive components in human milk. 3-Fucosyllactose (3-FL) is an abundantly present hMO that can be produced in sufficient amounts to allow application in infant formula. Lacto-N-triaose II (LNT2) can be obtained by acid hydrolysis of lacto-N-neotetraose (LNnT). Both 3-FL and LNT2 have been shown to have health benefits, but their impact on infant microbiota composition and microbial metabolic products such as short-chain fatty acids (SCFAs) is unknown. To gain more insight in fermentability, we performed in vitro fermentation studies of 3-FL and LNT2 using pooled fecal microbiota from 12-week-old infants. The commonly investigated galacto-oligosaccharides (GOS)/inulin (9 : 1) served as control. Compared to GOS/inulin, we observed a delayed utilization of 3-FL, which was utilized at 60.3% after 36 h of fermentation, and induced the gradual production of acetic acid and lactic acid. 3-FL specifically enriched bacteria of Bacteroides and Enterococcus genus. LNT2 was fermented much faster. After 14 h of fermentation, 90.1% was already utilized, and production of acetic acid, succinic acid, lactic acid and butyric acid was observed. LNT2 specifically increased the abundance of Collinsella, as well as Bifidobacterium. The GOS present in the GOS/inulin mixture was completely fermented after 14 h, while for inulin, only low DP was rapidly utilized after 14 h. To determine whether the fermentation might lead to enhanced colonization of commensal bacteria to gut epithelial cells, we investigated adhesion of the commensal Lactobacillus plantarum WCFS1 to Caco-2 cells. The fermentation digesta of LNT2 collected after 14 h, 24 h, and 36 h, and GOS/inulin after 24 h of fermentation significantly increased the adhesion of L. plantarum WCFS1 to Caco-2 cells, while 3-FL had no such effect. Our findings illustrate that fermentation of hMOs is very structure-dependent and different from the commonly applied GOS/inulin, which might lead to differential potencies to stimulate adhesion of commensal cells to gut epithelium and consequent microbial colonization. This knowledge might contribute to the design of tailored infant formulas containing specific hMO molecules to meet the need of infants during the transition from breastfeeding to formula