大孔树脂对苹果渣中多酚物质的吸附研究

通过静态吸附与解吸实验对8种大孔树脂进行筛选,结果表明:NKA-9型大孔树脂表现出最好的吸附性能与解吸效果,是较好的富集纯化苹果多酚树脂。通过单因素实验，确定NKA-9树脂的较佳动态吸附条件为苹果多酚液浓度2.50 mg/mL，pH 4.5，流速1.0 mL/min，较佳洗脱条件为乙醇浓度60%,洗脱流速1.0 mL/min,洗脱体积10 BV。     

Structural identification of novel oligosaccharides produced by Lactobacillus bulgaricus and Lactobacillus plantarum

β-Galactosidases (β-Gal) of lactic acid bacteria produce oligosaccharides from lactose when suitable acceptor carbohydrates are present. This study aimed to elucidate the structure of oligosaccharides formed by galactosylation of N-acetylglucosamine (GlcNAc) and fucose. Crude cellular extract of Lactobacillus bulgaricus and LacLM of Lactobacillus plantarum were used as sources of β-Gal activity. Disaccharides obtained by galactosylation of GlcNAc were identified as Gal-β-(1→4)-GlcNAc or Gal-β-(1→6)-GlcNAc by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and comparison with external standards. Trisaccharides were identified as Gal-β-(1→6)-Gal-β-(1→[4 or 6])-GlcNAc by LC-MS, analysis of the MS/MS spectra of selected in-source fragment ions, and their relative retention times. LC-MS analysis revealed the presence of five galactosylated fucosides, but their linkage type could not be identified, partly due to the lack of reference compounds. β-Gal of lactic acid bacteria may serve as suitable tools for the chemoenzymatic synthesis of therapeutic oligosaccharides.     

苹果渣多酚提取工艺的优化

为优化苹果渣中多酚提取工艺,并得到提取工艺和多酚组成之间的关系,利用微波辅助提取法设计了由物料颗粒、液料比、乙醇浓度、微波功率和微波提取时间5个因素构成的多酚提取优化试验流程,构建了涵盖黄酮、原花青素2种典型多酚物质和抗氧化能力的提取工艺评价指标体系。采用证据理论对不同工艺在评价指标下的焦元进行识别,并基于信度函数和似真函数得到了不同提取工艺的效用区间和优化方案。优化结果为：物料颗粒60目,液料比30?mL/g,乙醇体积分数60%,微波功率600?W,提取时间70?s,此条件下苹果渣多酚的提取量为213.83?mg/100g,黄酮提取量为83.21?mg/100g,原花青素提取量为52.79?mg/100g,抗氧化的EC50值为3.71?mg/100mL,验证了采用证据理论进行苹果渣多酚提取工艺优化的有效性。     

由苹果渣制备果胶低聚糖的工艺

为了促进苹果渣高增值利用,该研究探索了由苹果渣制备果胶低聚糖的工艺条件。首先以正交试验选取果胶提取工艺参数,再通过单因素和响应面试验,以产量为指标优化了果胶酶(EC232-883-6)水解苹果果胶制备果胶低聚糖的工艺。结果表明,温度90℃,提取时间90min,pH值1.8条件下提取,结合专利技术脱色制备了白色细粉状苹果果胶,得率11%,其多聚半乳糖醛酸含量84.3%。酶解制备低聚糖的最佳工艺条件为:pH值4.91,酶用量0.082U?mL-1,底物浓度8.5μmol?mL-1,反应温度25℃,反应时间14.94min,最佳条件下果胶低聚糖相对于果渣的得率为5.92%,产品较强地抑制了2种肠道有害菌——大肠杆菌和金黄色葡萄球菌的生长,对几种供试霉菌作用相对较弱。     

联合提取分离苹果渣果胶和多酚工艺优化

为研究从苹果渣中联合提取和分离果胶和多酚的最优工艺条件,该文利用酸提法从干燥的苹果渣中提取果胶和多酚,并利用疏水性大孔树脂进行分离。结合一种基于区间数多决策属性的评价方法优化提取分离过程。提取参数为果胶的产量和多酚的产量;分离参数包括:多酚吸附率、果胶吸附率和果胶的色泽变化。经优化后,提取的最优性条件为:pH值2.0,温度95℃,提取时间1.5h和料液比1:20(g/mL)。果胶、多酚物质的平均提取得率为196.49和10.98mg/g,分别占单一提取产率的92.28%和85.92%。经优化得出:最佳的大孔树脂是XDA-5。根据最优分离条件可得到提取液中98.32%果胶和92.15%多酚。结果表明,多属性决策理论的评价方法应用在提取分离苹果渣是可行的,可减少试验量。     

微波干燥预处理对苹果渣提取果胶的影响

利用微波干燥技术对湿苹果渣进行预处理,研究了微波干燥预处理对苹果渣提取果胶的影响.以探索一种新的干燥方法和提高果胶品质.结果表明:微波能量越大,苹果渣组织结构破碎越严重,其复水能力越弱;但对苹果渣颜色无显著影响;微波干燥预处理能显著提高果胶得率,微波预处理条件为300 W、15 min、7.5×10-2kW·h或500 W、10 min、8.3×10-2kW·h时微波预处理苹果皮渣的果胶提取得率比对照提高了50%左右,是较为适宜的微波处理条件,但微波能量大于8.3×10-2kW·h时,对果胶得率有不利影响;微波干燥预处理对果胶半乳糖醛酸含量无显著性影响,但显著提高了果胶的酯化度、固有黏度和黏均分子质量.     

Use of the 1H nuclear magnetic resonance spectra signals from polyphenols and acids for chemometric characterization of cider apple juices

The low field region (5.8-9.0 ppm) corresponding to aromatic protons and the region 1.8-3.0 ppm of the (1)H NMR spectra were used for characterization and chemometric differentiation of 52 apple juices obtained from six cider apple varieties. The data set consisted of 14 integrated areas corresponding to resonances from acids and phenolic compounds. Multivariate procedures based on hierarchical cluster and discriminant analysis were performed on selected signals of the spectra to determine whether it was possible to distinguish the different juices. Cluster analysis was able to satisfactorily classify the six apple varieties. Discriminant analysis, by means of stepwise procedure for variables selection and leave-one-out for cross-validation, was applied to 40 samples from the year 2001, obtaining recognition and prediction abilities of 100%. The most discriminant variables corresponded to poliphenols, (-)-epicatechin, phloridzin-phloretin, and p-coumaric, chlorogenic, and malic acids. The classification model was applied to 12 samples from apples harvested in the years 2002 and 2003, and the prediction ability was 91.7%.     

莲子酶解动力学分析与酶解产物多糖的表征

为开发更温和与简便的高纯度植物多糖新型酶法水解提取工艺,该研究以胃蛋白酶（内肽酶）和曲蛋白酶（端肽酶）作为复合酶,建立酶解过程的动力学模型。对上述工艺所提取制备的多糖与45 ℃水提法、90 ℃水提法、胃蛋白酶提取法所制得的莲子多糖进行营养成分分析与结构（单糖组成、红外光谱、热特性、低场核磁和动态热机械）表征。结果显示,单酶（胃蛋白酶）/双酶分段酶解的动力学模型分别为：D_H=2.101ln[1+(0.6133(E₀/S₀)+0.1441)t]和D''_H-D_H1=2.439ln[1+(3.923(E₀/S₀)+1.1756)t];双酶法提取的莲子多糖中多酚浓度（（0.42±0.008）mg/mL）和糖醛酸含量（15.65%±0.98%）最高,而以双酶法制得的莲子多糖蛋白质含量（0.73%±0.24%）最低;4种莲子多糖均含有葡萄糖 (Glc)、阿拉伯糖（Arab）、甘露糖（Man）和鼠李糖（Rha）,其中双酶法提取的多糖中半乳糖醛酸（Gal-UA）和Man的含量较多,分别为10.700%和10.752%;4种多糖均为α-型吡喃糖;双酶提取法相比水提法可有效降低莲子多糖中的蛋白质含量和玻璃化温度,提高结合水含量和亲水能力。酶解动力学模型可为莲子多糖纯化机制提供有效参考,尤其是双酶分段酶解法的特异性强、步骤简便,有利于提取和纯化莲子多糖成分,该研究可为动植物非淀粉类多糖的高质量提取和工业化生产提供理论基础。     

苹果渣中不同极性多酚的分离及体外抗氧化活性研究

为了优化苹果多酚提取工艺,进而提高苹果多酚提取物中强效多酚的含量,该文采用液-液萃取方法按极性大小对果渣多酚进行分离,进而采用1,1-二苯基-2-苦苯肼自由基法和铁离子还原/抗氧化力法对不同极性多酚体外抗氧化活性进行评价.结果表明:液-液萃取方法,可以实现不同极性多酚的有效分离,不同极性苹果多酚,其体外抗氧化活性有显著差异,水层多酚活性最强,其次为正丁醇层,乙酸乙酯层相对较弱,即多酚的抗氧化性与其极性呈正相关.因此,工业化提取苹果渣中多酚时,增大提取溶剂的极性有利于强效多酚的获取.     

Production of L-lactic acid and oligomeric compounds from apple pomace by simultaneous saccharification and fermentation: a response surface methodology assessment

The potential of apple pomace for lactic acid production by simultaneous saccharification and fermentation (SSF) was evaluated. The effects of the cellulase to solid ratio (CSR), the liquor to solid ratio (LSR), and the beta-glucosidase to cellulase ratio (BCR) on the kinetics of lactic acid generation were assessed, and a set of mathematical models was developed to reproduce and predict the lactic acid concentration of fermentation broths. Operating at low cellulase and cellobiase charges (1 FPU/g and 0.25 IU/FPU, respectively) and short reaction times (10 h), SSF media containing 27.8 g of lactic acid/L were obtained with a volumetric productivity of 2.78 g/Lh. Material balances showed that the SSF processing of 100 kg of dry apple pomace results in the production of 36.6 kg of lactic acid, 18.3 kg of oligomeric carbohydrates (which can be used as ingredients for functional foods), 8.4 kg of microbial biomass, and 8 kg uronic acids.     

Physiological effects of extraction juices from apple, grape, and red beet pomaces in rats

In comparison to classical fruit juice processing, polyphenols and dietary fiber can be extracted from pomace by means of pectinases and cellulases. In the present study, rats were fed with such produced extraction juices from apples, grapes, and red beets as drinking fluids instead of water for 4 weeks to evaluate their physiological effects. In all test groups, the intake of extraction juices was greater as compared to control (water intake), resulting in a higher urine excretion. In the apple and grape group, pH values in feces was lower than control. Administration of extraction juices from apples increased fecal counts of Lactobacillus and Bifidobacterium. More acetate and total short-chain fatty acids appeared in intestinal contents of the apple and red beet group. Furthermore, the intestinal contents of test groups contained higher concentrations of primary bile acids, cholesterol, and cholesterol metabolites but lower concentrations of secondary bile acids. The total amount of steroids excreted by these groups was also greater than control. Quercetin and isorhamnetin appeared in urine of rats fed extraction juices from apples and grapes; in urine of the former group, phloretin was found also. Administration of the extraction juices, enriched in secondary plant metabolites and dietary fiber, resulted in beneficial nutritional effects in rats.     

Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria

New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied. Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple. All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested. Bacteroides spp. are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides. Bifidobacterium spp. are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides. Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro. These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains.     

黑曲霉固态发酵苹果渣产木聚糖酶的工艺优化研究

为了获得有较高酶活值的低成本木聚糖酶产品,试验以苹果渣和棉粕为基料研究了黑曲霉(Aspergillus niger SL-05)固态发酵产木聚糖酶的最佳条件.初步试验结果表明:以棉粕和苹果渣(1:1)作为基础氮源和碳源得到的发酵曲的酶活值最高.通过Plackett-Burman试验筛选出了对产酶影响显著的3个因素:尿素、KH2PO4、含水率.进一步试验采用二次回归正交旋转试验设计研究了自变量(尿素、KH2PO4、含水率)对黑曲霉SL-05产酶的影响,通过响应面分析获得了产酶的最佳条件.模型的方差检验显示总回归达到了显著水平(P＜0.05),失拟性检验不显著,说明模型适合并且预测尿素、KH2PO4、含水率对产酶的影响非常有效.在最佳条件(基料棉粕和苹果渣比例为1:1,第2氮源尿素2.6%,无机盐KH2PO4 0.09%,速效碳源葡萄糖2%,含水率62.9%,30℃培养60 h)下,分别获得T5662、30000 U/g的木聚糖酶和纤维素酶的高酶活发酵干曲.以廉价的工农业废料作为基本培养基获得了有较高酶活的产品,经济优势明显.     

Purification and characterization of a hexanol-degrading enzyme extracted from apple

An enzyme having activity toward n-hexanol was purified from apple, and its biochemical characteristics were analyzed. The purification steps consisted of sedimentation with ammonium sulfate, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephadex G-100 column. The obtained enzyme had a yield of 16.00% with a specific activity of 18879.20 U/mg protein and overall purification of 142.77-fold. The enzyme showed activity to isoamylol, 1-propanol, n-hexanol, and isobutanol but not toward methanol and ethanol. With n-hexanol as a substrate, the optimum conditions were pH 4.0 and 30 °C for enzyme activity and pH 3.0-4.0 and temperatures below 40 °C for enzyme stability. The enzyme activity was increased significantly by adding l-cysteine and Fe(2+) at all tested concentrations and slightly by Zn(2+) at a high concentration but decreased by additions of EDTA, Ga(2+), K(+), Mg(2+), sodium dodecyl sulfate (SDS), sodium aluminum sulfate (SAS), dithiothreitol (DTT), and glutathione (GSH). The enzyme activities toward n-hexanol and n-hexanal were increased by NADH but decreased by NAD(+), in contrast to a decrease toward n-hexane by addition of both NAD(+) and NADH.     

Enzymatic production and characterization of konjac glucomannan oligosaccharides

Enzymes from a balanced human gut flora are promising tools to design prebiotic oligosaccharides. In this study, we investigated the action of enzymes from fecal bacteria on the complex polysaccharide konjac glucomannan (KGM). The oligosaccharides produced were compared to oligosaccharides from KGM digests with fungal endo-β-(1,4)-glucanase (EG) or endo-β-(1,4)-mannanase (EM). For this purpose, the oligosaccharides from the different digests were first studied for their structural characteristics like monosugar composition and exo-enzymatic degradability, as monitored by capillary electrophoresis with laser-induced fluorescence detection. Whereas the oligosaccharides produced by EG and EM were characteristic for the selectivity of the respective enzyme in cleaving the mannose-/glucose-sugar linkages of KGM, oligosaccharides produced by the fecal enzymes did not point to a sugar-selective degradation. The oligosaccharide fragments from the different digests indicated the KGM polysaccharide to be composed of a backbone composed of short mannose and glucose sequences, to which branches rich in mannose are attached.     

Structural characterization and biological activities of an exopolysaccharide kefiran produced by Lactobacillus kefiranofaciens WT-2B(T)

Lactobacillus kefiranofaciens, isolated from kefir grains, produces an extracellular polysaccharide when cultured, not only in PYG10 medium but also in a liquid medium containing a rice hydrolysate that had been previously degraded by treatment with a glucoamylase. The maximum yield of the polysaccharide, using the rice hydrolysate as the medium, was 2.5 g/L after a 7-day culture period at pH 5.0 and 33 degrees C. Compositional analysis, methylation analysis, specific rotation, and (1)H and (13)C NMR spectroscopy revealed that the structures of polysaccharides obtained from these two different culture media are essentially identical. The polysaccharide is composed of a hexasaccharide repeating unit and, thus, is known as kefiran. The weight-average molecular weight and the z-average radius of gyration of a sample, purified from the rice hydrolysate medium, were determined to be 7.6 x 10(5) g/mol and 39.9 nm, respectively, by gel permeation chromatography equipped with a multiangle laser-light-scattering photometer. Changes in blood pressure and serum components were examined in SHRSP/Hos rats, using doses of 100 and 300 mg of kefiran/kg of rat. A suppression in the increase in blood pressure was observed in these rats after 30 days. This activity is discussed in terms of the concentration of serum components of the rat, with emphasis on lipid components such as cholesterols, triglycerides, and free fatty acids.     

Variability of the polyphenolic composition of cider apple (Malus domestica) fruits and juices

Five French cider apple varieties were compared on the basis of their detailed polyphenol profile in the cortex and in the juices. Among the factors studied, variety was the most important variability factor in fruits, whereas polyphenol profiles showed an overall stability from one year to another, and a limited decrease of polyphenol concentration was observed during the starch regression period of fruit maturation. In juices, procyanidins remained the preponderant polyphenol class with concentrations up to 2.4 g/L even in centrifuged juices. Compared to the fruits, the average degree of polymerization of procyanidins was significantly reduced in the juice. Centrifugation of the crude juice had only minor effects on the polyphenol composition. For one variety, highly polymerized procyanidins with average degrees of polymerization of 25 were shown to be soluble in the centrifuged juice at a concentration of close to 1.2 g/L. Oxygenation of the juices during processing resulted in a significant decrease of all classes of native polyphenols. Catechins and procyanidins were particularly affected by oxidation, whereas caffeoylquinic acid was partly preserved. The transfer of polyphenols after pressing was maximal for dihydrochalcones and minimal for procyanidins with extraction yield values close to 80 and 30%, respectively.     

Agrochemical characterization,net N mineralization,and potential N leaching of composted olive‐mill pomace currently produced in southern Spain

In S Spain, the Andalusian olive oil industry generates annually 2.5–3.0 million tons of olive mill pomace, a by‐product which is comprised of the residues from the two‐phase oil‐extraction process. The agricultural policies of the EU have led to widespread interest in recycling these agricultural by‐products. Olive mill pomace might be evaluated as an organic fertilizer after composting, however, before wider use of composted olive mill pomace is advocated, characterization of the final product is needed. In this study, the physico‐chemical characteristics, net N mineralization, and the potential for N leaching of 7 out of the 11 olive‐mill‐pomace composts currently produced in the Andalusian olive mills were investigated. Compost of olive mill pomace differed in the proportions of raw materials co‐composted with the olive mill pomace, such as olive leaf material, manure, and straw. In all the composts tested, organic matter, total C and K were high with 60.5%, 30.7%, and 1.7% on average, respectively, whereas total P was low (0.4%) and with intermediate levels of N (1.5%). Compost pH (8.03), electrical conductivity (2.85 dS m^–1), and germination index (65% on average) were adequate for agricultural use. Furthermore, principal component analyses revealed a clear relationship between the quality of the composts and the proportion of manure mixed with the raw materials. Net N mineralization was negative on average (–20 μg IN g^–1) after 1 y, but positive after 2 y of incubation with up to 94% of available N from the total N added and the short‐term potential N leaching after compost application was negligible (less than 3.9% of added N) and much lower than the other N fertilizer with up to 80% added N leached. Overall, results of this study clearly show that these currently produced composts of olive mill pomace are suitable as soil improvers for agricultural purposes, but may not contribute significantly as a N fertilizer for up to 2 y after application.     

Chemical characterization and anti-inflammatory effect of polysaccharides fractionated from submerge-cultured Antrodia camphorata mycelia

Five polysaccharide fractions (AC-1, AC-2, AC-3, AC-4, and AC-5) were obtained after systemic solvent extractions and precipitations from Antrodia camphorata mycelia with yields of 2.92, 10.38, 1.65, 0.34, and 1.64%, respectively. Gel permeation chromatography (GPC) analysis showed that the distribution of mean molecular mass of the fractionated polysaccharides was in the range of 394-940 kDa. The proximate compositions from each polysaccharide fraction revealed that all fractions belonged to the category of glycoprotein, having ratios of carbohydrate/protein ranging from 0.29 to 10.79 (w/w). Glucose or galactose was the major monosaccharide in all fractions except fraction AC-2, which has a mean molecular mass of 394 kDa with lyxose as the most prominent constituent. In the evaluation of the DPPH* radical scavenging capability, fraction AC-1 and AC-2 polysaccharides showed the better capabilities, around 74.5 and 50.5%, respectively, compared to the reference control of Trolox (87.5%) at a concentration of 1 microM. In testing with macrophage RAW264.7 cells, fraction AC-2 demonstrated a rather potent anti-inflammatory capability. Furthermore, the lipopolysaccharide-induced NO production and the protein expression by the inducible nitric oxide synthase (iNOS) gene were inhibited, respectively, in a dose-dependent (50-200 microg/mL) manner by fraction AC-2 polysaccharide.     

Structural and biological study of carboxymethylated Phellinus linteus polysaccharides

Polysaccharides isolated from Phellinus linteus were chemically modified by carboxymethylation, and the structural and physiological properties of the derivative were investigated. 13C NMR spectroscopy showed that the polysaccharides extracted from P. linteus contained (1-3)-beta-glucans with a (1-6)-linkage. The carboxymetehylation of the P. linteus polysaccharides was confirmed by Fourier transform infrared spectroscopy, and the degree of substitution was obtained by the potentiometric titration, which was calculated to be 0.63. The bronchoalveolar lavage experiments showed that the carboxymethylated derivative raised the nitric oxide production. In addition, the carboxymethylation stimulated in vitro cytotoxic activity against the HT1080 cell line. Thus, the derivative exhibited the enhanced activity of immune systems, which would be explained by the improved water solubility and structural changes by carboxymethylation. However, a slight decrease in the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity of the derivative was observed.