Extraction of green labeled pectins and pectic oligosaccharides from plant byproducts

Green labeled pectins were extracted by an environmentally friendly way using proteases and cellulases being able to act on proteins and cellulose present in cell walls. Pectins were isolated from different plant byproducts, i.e., chicory roots, citrus peel, cauliflower florets and leaves, endive, and sugar beet pulps. Enzymatic extraction was performed at 50 degrees C for 4 h, in order to fulfill the conditions required for microbiological safety of extracted products. High methoxy (HM) pectins of high molar mass were extracted with three different enzyme mixtures. These pectins were subsequently demethylated with two pectin methyl esterases (PMEs), either the fungal PME from Aspergillus aculeatus or the orange PME. It was further demonstrated that high molar mass low methoxy (LM) pectins could also be extracted directly from cell walls by adding the fungal PME to the mixture of protease and cellulase. Moreover, health benefit pectic oligosaccharides, the so-called modified hairy regions, were obtained after enzymatic treatment of the residue recovered after pectin extraction. The enzymatic method demonstrates that it is possible to convert vegetable byproducts into high-added value compounds, such as pectins and pectic oligosaccharides, and thus considerably reduce the amount of these residues generated by food industries.     

Solvent effects on the molecular properties of pectins

Measurements revealed that LiAc/HAc buffer, when compared with other solvents, gave relatively low values of turbidity for five commercial pectins with various apparent molar masses and degrees of methyl esterification (DE). Therefore, HPSEC with on-line light scattering and viscosity detection was employed to compare LiAc/HAc buffer against NaNO(3) solution, a commonly used mobile phase for measuring the molecular properties of these pectins and an additional pectin prepared by microwave extraction. Microwave-extracted pectin was included in the study for its higher molar mass and DE compared with commercial pectins. Most commercial samples were more soluble and had a higher molar mass when dissolved in NaNO(3) than in LiAc/HAc buffer, whereas the microwave-extracted pectin was more soluble in LiAc/HAc buffer and had about the same molar mass. Furthermore, association fragments of pectin contained in samples were more dissociated by LiAc/HAc buffer than by NaNO(3). For the samples studied, weight-average molar masses ranged from about 41000 to 307000, weight-average intrinsic viscosities from about 0.86 to 9.76 dL/g, z-average radii of gyration from about 13 to 45 nm, and Mark-Houwink constants from about 0.62 to 0.94.     

Physical and oxidative stability of fish oil-in-water emulsions stabilized with beta-lactoglobulin and pectin

The oxidation of fatty acids can be inhibited by engineering the surface of oil-in-water emulsion droplets to decrease interactions between aqueous phase prooxidants and lipids. The objective of this research was to evaluate whether emulsions stabilized by a multilayer emulsifier systems consisting of beta-lactoglobulin and citrus or sugar beet pectin could produce fish oil-in-water emulsions that had good physical and oxidative stability. Sugar beet pectin was compared to citrus pectin because the sugar beet pectin contains the known antioxidant, ferulic acid. A primary Menhaden oil-in-water emulsion was prepared with beta-lactoglobulin upon which the pectins were electrostatically deposited at pH 3.5. Emulsions prepared with 1% oil, 0.05% beta-lactoglobulin, and 0.06% pectins were physically stable for up to 16 days. As determined by monitoring lipid hydroperoxide and headspace propanal formation, emulsions prepared with the multilayer system of beta-lactoglobulin and citrus pectin were more stable than emulsions stabilized with beta-lactoglobulin alone. Emulsions prepared with the multilayer system of beta-lactoglobulin and sugar beet pectin were less stable than emulsions stabilized with beta-lactoglobulin alone despite the presence of ferulic acid in the sugar beet pectin. The lower oxidative stability of the emulsions with the sugar beet pectin could be due to its higher iron and copper concentrations which would produce oxidative stress that would overcome the antioxidant capacity of ferulic acid. These data suggest that the oxidative stability of oil-in-water emulsions containing omega-3 fatty acids could be improved by the use of multilayer emulsion systems containing pectins with low metal concentrations.     

Role of protein and ferulic acid in the emulsification properties of sugar beet pectin

The ability of sugar beet pectin to stabilize 20% w/w limonene oil-in-water emulsions has been investigated. The size of the oil droplets as determined by laser diffraction measurements decreased from about 15 mum to about 6 mum when the pectin concentration increased from 0.05% to 2% w/w but leveled off thereafter, suggesting complete coverage of the oil droplets by the polymer at this optimum concentration. Isotherms for the adsorption of pectin, protein, and ferulic acid were constructed. The adsorption capacities at the oil-water interface of approximately 1.4 and approximately 0.2 mg/m (2) for protein and ferulic acid, respectively, compared to approximately 9.5 mg/m (2) for pectin revealed that the adsorbed fractions of the pectin sample were rich in protein (14.7%) and ferulic acid (2.1%) given that there were only 2.7% protein and 1.06% ferulic acid present in the whole pectin sample. Direct measurements on the adsorbed fraction recovered from the oil droplets via desorption with SDS confirmed that it contained 11.1% protein and 2.16% ferulic acid. The results suggest that one or both of these two functional groups adsorb onto the surface of the oil droplets and stabilize the emulsions. High molecular mass fractions adsorbed preferentially onto oil droplets during emulsification. As compared to those made with gum arabic, the emulsion samples made with sugar beet pectin samples exhibited similar (or even slightly higher) stability.     

Cellulase and protease preparations can extract pectins from various plant byproducts

The use of protease and cellulase preparations to extract pectins from plant byproducts (chicory, cauliflower) was investigated. Different enzymatic preparations were characterized by their activities toward proteins, cellulose, and pectins. These preparations were then tested regarding pectin extraction, and extraction conditions (nature and concentration of enzyme, incubation time) were optimized. Enzymatic and acidic extractions were compared and also combined in sequential extractions. This study shows that it is possible to extract pectins by using cellulases and proteases. Enzymes can extract pectins with a higher yield ( approximately 35%) than acid (approximately 28%) but enzyme-extracted pectins have a smaller molar mass (300,000 g/mol) than acid-extracted pectins (500,000 g/ mol). Different hypotheses are tested and discussed to explain this mass difference.     

微波辅助提取黄皮果肉果胶工艺参数优化

为优化微波辅助提取黄皮果肉中果胶的工艺,采用均匀设计法,考察了微波功率、提取时间、液料比及提取液pH值4个因子对黄皮果胶得率的影响。利用SAS软件对试验数据进行模型拟合和回归分析,确定提取时间和液料比是影响果胶得率的重要因子,并最终获得微波辅助提取黄皮果胶的最优工艺参数为:微波功率600W,提取时间8min,液料比24:1mL/g,提取液pH值2.0。在此条件下果胶得率为3.59%。通过对果胶基本特性的测定分析,该工艺提取的黄皮果胶品质基本符合国家标准,可为黄皮果胶提取的工业化放大提供参考。     

Feruloylated and nonferuloylated arabino-oligosaccharides from sugar beet pectin selectively stimulate the growth of Bifidobacterium spp. in human fecal in vitro fermentations

The side chains of the rhamnogalacturonan I fraction in sugar beet pectin are particularly rich in arabinan moieties, which may be substituted with feruloyl groups. In this work the arabinan-rich fraction resulting from sugar beet pulp based pectin production was separated by Amberlite XAD hydrophobic interaction and membrane separation into four fractions based on feruloyl substitution and arabino-oligosaccharide chain length: short-chain (DP 2-10) and long-chain (DP 7-14) feruloylated and nonferuloylated arabino-oligosaccharides, respectively. HPAEC, SEC, and MALDI-TOF/TOF analyses of the fractions confirmed the presence of singly and doubly substituted feruloylated arabino-oligosaccharides in the feruloyl-substituted fractions. In vitro microbial fermentation by human fecal samples (n = 6 healthy human volunteers) showed a selective stimulation of bifidobacteria by both the feruloylated and the nonferuloylated long-chain arabino-oligosaccharides to the same extent as the prebiotic fructo-oligosaccharides control. None of the fractions stimulated the growth of the potential pathogen Clostridium difficile in monocultures. This work provides a first report on the separation of potentially bioactive feruloylated arabino-oligosaccharides from sugar beet pulp and an initial indication of the potentially larger bifidogenic effect of relatively long-chain arabino-oligosaccharides as opposed to short-chain arabino-oligosaccharides.     

Use of a Plackett-Burman experimental design to examine the impact of extraction parameters on yields and compositions of pectins extracted from chicory roots (Chicorium intybus L.)

Chicory root pectin was isolated by acid extraction followed by alcohol precipitation. Because the extraction conditions have important effects on the features of pectins, an experimental design was used to study the influence of 17 different extraction parameters on yield and composition of pectin: pH, temperature, time of extraction, solid/liquid ratio, and different pretreatments of the pulps before extraction. Twenty extractions were conducted and examined for their significance on yield and sugar content using the Plackett-Burman factorial design. The acid extraction of chicory roots resulted in an average yield of 11% containing 86% of sugars. It was found that extraction temperature, time, protease pretreatment, water purity, and water washing of pulps significantly affected yield and pectin composition with an increase of yield and purity of pectin in harsher extraction conditions.     

Gastrointestinal Release of β‐Glucan and Pectin Using an In Vitro Method

The release of soluble dietary fiber is a prerequisite for viscous effects and hence beneficial health properties. A simple in vitro method was adapted to follow the release during gastrointestinal digestion, and the percentage of solubilized fiber was measured over time. β‐Glucan from oat bran was mainly released during gastric digestion while the release of pectin from sugar beet fiber continued in the small intestine. Unmilled fractions of sugar beet fiber released more soluble fiber than oat bran flakes, probably due to the porous structure of sugar beet fiber as a result of manufacturing processes, but also due to differences in source. Milling to smaller fiber particles significantly improved releasability (from 20 to 55% released β‐glucan and from 50 to 70% released pectin, respectively, after digestion). When milled fibers were included in individual food matrices, the release was reduced by protein and starch matrices (5% β‐glucan and 35% pectin released, respectively) and slowed by fat (45% β‐glucan and 60% pectin released). This may result in a too low or too late release in the upper small intestine to be able to interfere with macronutrient uptake. The method may be suitable for predicting the gastrointestinal release of soluble dietary fibers from food matrices in the development of healthy food products.     

不同柑橘类果胶分子的理化特性、结构及凝胶性能

为了解不同果胶的理化性质、结构与其功能特性的区别,拓展不同性能果胶的针对化应用,提高柑橘加工副产物的附加值。该研究采用酸提法从上饶柚子皮、赣南脐橙皮、南丰蜜桔皮、日本柚子皮、小青柑皮、柠檬皮6种柑橘类水果果皮渣中分别提取果胶,依次为HMP-1、HMP-2、HMP-3、HMP-4、HMP-5和HMP-6,并对其理化特性、结构及凝胶性能进行比较分析。结果表明：不同来源的6种柑橘果胶酯化度在67.20%～75.45%,均为高酯果胶（high methoxyl pectin, HMP）,具备阴离子多糖特性,在提取过程中均出现了不同程度的降解,理化特性和分子结构上存在一定差别。果胶的分子量、黏度、分子链结构等因素共同作用于果胶的凝胶性能,6种HMP均能形成具有一定强度的酸/糖凝胶,将水分截留在三维凝胶网络中,经历冻融循环过程持水性仍在96%以上,具备较好的稳定性。其中分子量较低的HMP-1和HMP-6凝胶强度最高,胶凝度均在200°以上,这与HMP-1的高同型半乳糖醛酸聚糖（HG）含量、较完整的分子结构,以及HMP-6较高的鼠李糖半乳糖醛酸聚糖-I（RG-I）含量及分支度有关,而分子量及黏度较高的HMP-5的弱凝胶强度则与其较大的分子链降解程度及低RG-I含量有关,表明HG和RG-I结构域对凝胶的形成均有促进作用。原子力显微镜图像也显示,HMP-1和HMP-6中出现较为明显的分子链交联。对不同柑橘类果胶的理化性质、结构与凝胶特性进行比较分析,为确定凝胶性能更加优异的提取原料、果胶的扩大化生产及针对性应用提供了理论参考。     

Citrus pectin: characterization and inhibitory effect on fibroblast growth factor-receptor interaction.

This study was undertaken to characterize the pectin from four citrus species and to determine their in vitro inhibitory activities on the binding of fibroblast growth factor (FGF) to the FGF receptor (FGFR). Pectin from various parts of lemon, grapefruit, tangerine, and orange were isolated and characterized. Tangerine had the highest pectin content among the four citrus species. Segment membrane contained as much as or more pectin than flavedo/albedo. Anhydrogalacturonic content was highest in pectin from segment membrane of tangerine and flavedo/albedo of grapefruit. Lemon pectin contained the highest methoxyl content (MC), and grapefruit contained the largest proportion of lower molecular weight (<10000 Da) pectin. Tangerine contained the highest neutral sugar in both flavedo/albedo and segment membrane. The interdependency of heparin on factor-receptor interaction provides a means for identifying new antagonists of growth factor activity and thus for treatment of various diseases. These results showed that pectin significantly inhibited the binding of FGF-1 to FGFR1 in the presence of 0.1 microg/mL heparin. The pectin from the segment membrane of lemon was the most potent inhibitor. The inhibition activity was significantly correlated with sugar content, MC, and size of pectin. Kinetic studies revealed a competitive nature of pectin inhibition with the heparin, a crucial component of the FGF signal transduction process. The observation that the heparin-dependent biological activity of FGF signal transduction is antagonized by citrus pectin should be further investigated for the use of these pectins as anti-growth factor agents for potential health benefits.     

六种果皮原料果胶的理化及凝胶特性比较

为了解不同品种水果的果皮(柚子皮、西番莲皮、脐橙皮、石榴皮、榴莲皮)以及向日葵盘所提取果胶的理化和质构特性,研究了不同原料果胶的得率、色泽、果胶酸含量、甲氧基含量、酯化度、黏度及质构特性,特别是采用高效液相色谱准确测定了各类果胶的分子量。结果表明:柚子皮、向日葵盘和脐橙皮果胶质量分数较高,分别为18.06%、14.61%和14.43%;西番莲皮果胶质量分数为8.76%;而石榴皮及榴莲皮果胶质量分数较低(均<3%)。从分子量看,石榴皮、脐橙皮果胶分子量较大(>1000kDa),向日葵盘果胶分子量最小(483kDa)。此外,几种果胶的溶胶均属低黏度值果胶(<25厘泊),且在pH值为7.0时黏度最大、在pH值为5.0时黏度最小。结合凝胶质构分析表明:石榴皮果胶分子量最大,凝胶强度最大,但为高甲氧基果胶,且得率较低;而向日葵盘果胶分子量最小,但得率较高,且为低甲基果胶,在非糖及含糖体系中均可形成性能优良的凝胶,因此是生产果胶的良好原料。该文为果胶的生产及应用提供参考。     

Enzymatic modification of pectin to increase its calcium sensitivity while preserving its molecular weight

A commercial high-methoxy citrus pectin was treated with a purified salt-independent pectin methylesterase (PME) isozyme isolated from Valencia orange peel to prepare a series of deesterified pectins. A series of alkali-deesterified pectins was also prepared at pH 10 under conditions permitting beta-elimination. Analysis of these pectins using high-performance size exclusion chromatography (HPSEC) with on-line multiangle laser light-scattering, differential viscometer, and refractive index (RI) detectors revealed no reduction in weight-average molecular weight (M(w); 150000) in the PME-treated pectin series, whereas a 16% reduction in intrinsic viscosity (IV) occurred below a degree of esterification (DE) of 47%. In contrast, alkali deesterification rapidly reduced both M(w) and IV to less than half of that observed for untreated pectin. PME treatment of a non-calcium-sensitive citrus pectin introduced calcium sensitivity with only a 6% reduction in the DE. Triad blocks of unesterified galacturonic acid were observed in (1)H nuclear magnetic resonance spectra of this calcium-sensitive pectin (CSP). These results demonstrate that the orange salt-independent PME isozyme utilizes a blockwise mode of action. This is the first report of the preparation of a CSP by PME treatment without significant loss of the pectin's M(w) due to depolymerization.     

Biodegradable composites from sugar beet pulp and poly(lactic acid)

Sugar beet pulp and poly(lactic acid) (PLA) composites were prepared by compression-heating. The resultant thermoplastics had a lower density, but they had tensile strength similar to that of pure PLA specimens as well as the same geometric properties. Tensile properties depended on the initial water content of sugar beet pulp and the process by which composites were manufactured. In comparison with sugar beet pulp, the composite showed improved water resistance. This can be attributed to the hydrophobic character of PLA and pulp-matrix interactions. The composite thermoplastics showed suitable properties for potential use as lightweight construction materials.     

Pectic methyl and nonmethyl esters in potato cell walls.

Because pectins are released from potatoes and other plants under conditions that cleave ester linkages, it has been suggested that there are other galaturonoyl ester cross-links between pectin chains in addition to the known non-cross-linking methyl esters. A microscale titration method and a copper binding method were developed for the measurement of total polymer carboxyl (essentially pectic) ester content in potato cell walls. Relative to the uronic acid content of the cell walls, the degree of total esterification was 57-58%. Comparison with levels of methanol released on ester hydrolysis allowed nonmethyl uronoyl esters to be estimated to be 14-15% relative to total uronic acid. The possibility of nonmethyl-esterified linkages being formed in potato cell walls by a side-reaction catalyzed by pectin methyl esterase (PME) was investigated, but no increase in nonmethyl-esterified pectin was observed under conditions where pectin was being effectively de-esterified by endogenous PME activity.     

灌溉制度对膜下滴灌甜菜产量及水分利用效率的影响

为制定新疆合理的甜菜膜下滴灌制度,设置3个灌水次数(8、9和10次)和2个灌水定额(45和60mm)两因素全组合试验,于2016—2017年在新疆玛纳斯县农科院甜菜改良中心开展田间试验。结果表明,灌水次数增加时甜菜叶面积指数与产量增加,含糖率降低,对甜菜的水分利用效率、耗水量无明显影响(P0.05),甜菜叶绿素值随灌水次数与定额增加呈下降趋势;在灌水次数与定额交互作用下,灌水8次时由于土壤相对含水率低于50%,甜菜会减产;当灌水9次,灌水定额为45 mm时,增加15 mm灌水定额土壤相对含水率达50%以上,此时甜菜增产7.4%～7.7%,糖产增加9.4%～9.7%;而继续增加灌水次数时,会导致甜菜含糖率降低而降低糖产。因此针对新疆膜下滴灌甜菜以60 mm灌水定额灌水9次为宜,可获得高产与糖产,较传统新疆膜下滴灌甜菜制度节水10%。该研究对指导新疆膜下滴灌甜菜灌溉制度具有一定意义。     

Interactions of soy protein fractions with high-methoxyl pectin

A protein-binding technique was employed to visualize, using scanning electron microscopy, the soy protein as well as the association between HMP and soy protein fractions. Image analysis indicated that at pH 7.5 and 3.5 soy protein isolate showed a bimodal distribution of sizes with an average [ d(0.5)] of about 0.05 microm, but at pH 3.8 the proteins formed larger aggregates than at high pH. Addition of HMP at pH 3.8 changed the surface charge of the particles from +20 to -15 mV. A small addition of HMP caused bridging of the pectin between soy protein aggregates and destabilization. With sufficient HMP, the suspensions showed improved stability to precipitation. The microscopy images are the first direct evidence of the interactions between soy proteins with high-methoxyl pectin (HMP).     

微波辅助提取桔皮果胶的理化及凝胶特性比较

为了增加桔皮利用率,提高桔皮中果胶得率和品质,以5种新鲜桔皮(丑橘、砂糖橘、芦柑、金橘、贡桔)为原料,采用微波辅助法提取桔皮中果胶,对果胶理化特性和凝胶特性进行研究。分析果胶理化及凝胶特性的相关性,并采用主成分分析法确定影响果胶凝胶特性的主要因素,优化微波辅助法提取工艺参数。结果表明:5种桔皮果胶都是高酯果胶,5种果胶理化及凝胶特性存在显著差异(P0.05),其中芦柑果胶得率和半乳糖醛酸质量分数最高,分别为12.36%、78.61%;且5种果胶理化及凝胶特性呈显著相关性(P0.05),主成分分析法确定甲氧基质量分数和果胶得率分别为主成分1和2中主要影响因子,甲氧基质量分数决定果胶种类,因此果胶种类和得率是桔皮果胶提取的主要因素。微波提取优化参数:以芦柑桔皮为原料,料液比为1:26 g/g,p H值1.30,微波功率440 W,微波时间62 s,果胶得率为16.00%。研究结果为桔皮果胶加工和应用提供理论指导。     

Elucidation of the emulsification properties of sugar beet pectin

A protocol has been developed to fractionate sugar beet pectin using hydrophobic affinity chromatography. Three samples eluted from the column using 4 M NaCl as solvent (fractions 1A, 1B, and 1C), two fractions eluted using 2 M NaCl (fractions 2A and 2B), and one fraction eluted using water (fraction 3). The fractions were shown to be very polydisperse, and differences between the GPC refractive index and UV absorbance (214 nm) elution profiles demonstrated chemical heterogeneity. They were found to contain significantly different proportions of protein (1A, 2.79%; 1B, 0.97%; 1C, 0.77%; 2A, 1.41%; 2B, 5.09%; and 3, 5.89%) and ferulic acid (approximately 1A, 0.5%; 1B, 0.5%; 1C, 0.9%; 2B, 1.5%; and 3, 2%). The weight-average molecular mass, M(w), of the fractions also varied (1A, 153 kDa; 1B, 155 kDa; 1C, 306 kDa; 2A, 562 kDa; 2B, 470 kDa; 3, 282 kDa). Three fractions, that is, 1A, 1B, and 3, produced orange oil emulsions with a relatively small droplet size that were stable over a period of weeks. The other three fractions (1C, 2A, and 2B with higher M(w) values) produced emulsions with an initially larger droplet size, and the droplet size increased considerably over time. The increased droplet size may be influenced by the viscosity of the aqueous continuous phase. There was no simple relationship between protein or ferulic acid content and emulsification ability. For example, fraction 1B, which contained the lowest proportion of both protein and ferulic acid, produced stable emulsions of similar droplet size to fraction 3, which contained the largest proportion of protein and ferulic acid. The role of protein in the emulsification process was investigated by measuring the amount of protein in the aqueous phase before and after emulsification. It was clearly demonstrated that proteinaceous material adsorbed at the oil-water interface. It is evident that the emulsification properties of sugar beet pectin are influenced by the accessibility of the protein and ferulic acid groups to the surface of the oil droplets, the proportion of ester groups, and the molecular mass distribution of the fractions.     

Addition of pectin and soy soluble polysaccharide affects the particle size distribution of casein suspensions prepared from acidified skim milk

Pectins are negatively charged polysaccharides employed as stabilizers in acidified milk dispersions, where caseins aggregate because of the low pH and serum separation needs to be prevented. The objective of this research was to study the effect of charge on the stabilizing functionality of the polysaccharide in acid milk drinks. Unstandardized pectins with various charges (as degree of esterification, DE) as well as soybean soluble polysaccharide (SSPS) were tested for their stabilizing behavior as a function of pH and concentration. Skim milk was acidified by glucono-delta-lactone and then homogenized in the presence of polysaccharide at different pH values (in the range from 4.2 to 3.0). Measurements of particle size distribution demonstrated that pectins with a DE of 71.4, 68.6, and 67.4 stabilized milk at pH > 4.0. Pectins with a lower DE (63.9%) needed a higher concentration (0.4%) at the same pH to show a monomodal distribution of particle sizes. Pectins with lower DE (<50%) did not stabilize the dispersions. Although this difference in behavior was attributed mainly to the pectin charge, the efficiency in stabilizing the casein dispersion decreased with decreasing pectin size. For example, the high methoxyl pectin (HMP) with 63.9 DE was smaller in size than the HMPs with a higher charge. Pectins showed a pH-dependent stabilization effect, as at pH < 4.0 the dispersions contained aggregates. When SSPS was used to stabilize acid milk, at pH < 4.0, it showed a better stabilization behavior than HMP. When SSPS and pectin were used in combination, the particle size distribution of the acid milk dispersion was pH-dependent, and results were similar to those for samples containing pectin alone. This suggested that in the mixture, pectin dominated the behavior over SSPS, even when an excess of SSPS was added to the dispersions before homogenization.