耐消化道逆环境双歧杆菌菌株A04体外促进生长因子的研究

研究了4种低聚糖、8种中药及4种食品原料体外对双歧杆菌菌株A04的增殖作用。结果表明,大豆低聚糖的增殖效果最明显（P〈0．01）,其次是低聚异麦芽糖和低聚果糖,水苏糖则几乎没有增殖效果,而且当改良MRS中低聚糖的浓度达到10％时,对其反而具有一定的抑制作用;8种中药在浓度为2％的时均有增殖效果,以Z3、Z1和Z2效果最显著（P〈0．01）,但其浓度不同对双歧杆菌菌株A04的增殖影响截然不同;本试验采用的4种食品原料对双歧杆菌均有促生长作用,较好的增殖作用的组合分别为：M-5％,CJ-2％,TE-2％和LE-5％。     

发酵法精制大豆低聚糖的研究

研究了三种面包酵母对大豆低聚糖碳源利用的选择性。结果表明 :面包酵母C可选择性地利用蔗糖 ,而水苏糖和棉子糖的保留率大于 96 %;通过添加酵母膏 ,经过 3 6h培养 ,面包酵母C可全部利用大豆低聚糖中的蔗糖。进一步研究表明 ,以大豆乳清废糖浆为原料直接发酵再经下游处理 ,可得到蔗糖含量低于 1 3 %的精制大豆低聚糖干粉。     

大豆多糖对双歧杆菌及人肠道菌群生长的影响

目的研究大豆多糖对双歧杆菌及肠道菌群生长的影响。方法替换Bs培养基中的碳源,分为不加糖、加葡萄糖2％、加大豆多糖2％、加大豆多糖5％、加低聚果糖2％五组,加3种双歧杆菌（长双歧、青春双歧、两歧双歧）菌液1％,测其24h后的活菌数,比较大豆多糖对双歧杆菌生长的影响;替换Bs培养基中的碳源,分为不加糖、加葡萄糖2％、加大豆多糖2％,加低聚果糖2％四组,加人体粪便菌液1％,模拟人体肠道环境厌氧培养24h后,用选择性培养基测其肠杆菌、肠球菌、双歧杆菌、乳酸杆菌的活菌数,观察大豆多糖对人体肠道菌群的影响。结果大豆多糖添加量为5％时对长双歧的促进作用明显优于不加糖组（P〈0．05）;大豆多糖对人体肠道各菌群的生长促进作用与低聚果糖差异无显著性（P〉0．05）。结论大豆多糖对长双歧杆菌的体外促进作用较明显;以粪菌群发酵糖试验表明,大豆多糖对乳杆菌和双歧杆菌均有促进作用,和低聚果糖作用效果相比差异无显著性（P〉0．05）,具有益生元的特性。     

大豆低聚糖对肠道菌群结构调节的研究

目的 :对山松牌大豆低聚糖对肠道菌群调节的效果进行研究。方法 :通过动物实验和人体试验。结果 :山松牌大豆低聚糖能增殖体内乳杆菌和双歧杆菌数量 ;在较高剂量时 ,能使体内的肠球菌和肠杆菌增殖。采用B/E值 (即双歧杆菌和肠杆菌的比值 )作为指标进行分析发现 ,该制品在低剂量时能更好地优化肠道菌群结构。结论 :大豆低聚糖对肠道菌有较好的调节效果     

异麦芽寡糖促进肠道分离菌生长的体外实验

目的 观察添加１％异麦芽寡糖对从正常人阑尾标本中分离双歧杆菌、乳杆菌和类杆菌的体外促生长作用。方法 分别在培养前和培养１２、２４、３６ｈ和４８ｈ,用７２１分光光度计和ｐＨ计测定培养液ＯＤ值和ｐＨ值变化。结果 接种双歧杆菌的培养液ＯＤ值从０．１２８增加到１．７,ｐＨ值由７．１降到４．２,增殖效果与添加１％葡萄糖相当。接种乳杆菌的培养液ＯＤ值从０．１７增加到０．８５,ｐＨ值由７．０降到４．８,增殖效果低于添加１％葡萄糖。接种类杆菌的培养液ＯＤ值从０．１０５增加到１．６５,ｐＨ值由７．１降到５．１,增殖效果高于添加１％葡萄糖。结论 添加１％民麦芽寡糖具有促进肠道分离菌双歧杆菌、乳杆菌和类杆菌生长的作用。     

低聚果糖体内外对肠道菌的影响

目的 :了解低聚果糖 (FOS)体内外对肠道菌双歧杆菌、类杆菌和肠杆菌科菌的增殖作用。方法 :FOS 1g/ (kg· bw· d) ,灌服 (ig)小鼠 ,连续 14 d后 ,取粪便 ,选择性培养基平板菌落计数法测各菌群。体外试验中 ,按 1%的 FOS添加到各人肠道分离菌培养液中 ,培养 2 4h后测其吸光度 A值和 p H值变化。结果 :ig FOS的小鼠肠道双歧杆菌和肠杆菌数量分别是 9.16± 0 .67和 8.3 3± 0 .70 (log10 N CFU/ g) ,高于对照组(P<0 .0 5)。 FOS体外对各肠道分离菌均有增殖作用 ,依大小分别是双歧杆菌、类杆菌和肠杆菌 ,其 A值分别增加了 0 .8～ 1.192、0 .80 2和 0 .198～ 0 .461,其 p H值分别降低了 1.5～ 1.68、1.2和 0 .2～ 0 .58。结论 :FOS体内外有相对选择性增殖双歧杆菌的作用 ,对类杆菌和肠杆菌也有调整作用     

功能性低聚糖生产工艺的研究现状

功能性低聚糖（functional oligosaccharides）是由2—10个单糖通过糖苷键连接形成直链或支链的一类寡糖,具有低热量、抗龋齿、抗肿瘤、防治糖尿病、防止腹泻和便秘等生理作用。因具有糖类某些共同特性,可作为甜食配料,但人体肠道内不具备分解消化功能性低聚糖的酶系统,所以不被人体胃酸、胃酶降解,不在小肠吸收,直接进入大肠内为双歧杆菌所利用,是一类优良的双歧杆菌增殖因子,故又名双歧因子。功能性低聚糖包括水苏糖、棉子糖、低聚果糖、低聚木糖、低聚半乳糖、大豆低聚糖和低聚异麦芽糖等。近年来,国外上市品种有10多种,生产批量较大,我国自1996年开始才有批量生产,主要以低聚异麦芽糖、低聚果糖和大豆低聚糖为主。本文综述了不同种类的功能性低聚糖及生产工艺的现状。     

植物组织中低聚糖乙酰化及毛细管气相色谱分析

建立一种用乙酰化衍生处理低聚糖并用毛细管气相色谱-FID进行分析的方法。以1-甲基咪唑为催化剂并以乙酸酐为乙酰化试剂,同时对植物样品中蔗糖、棉子糖和水苏糖等低聚糖乙酰化产物进行毛细管气相色谱分离和FID检测。确定了低聚糖乙酰化衍生物的毛细管气相色谱分析条件,并对低聚糖乙酰化反应条件及色谱分离条件进行了优化。结果表明,在80–1000ng·μL–1范围内线性关系良好,蔗糖、棉子糖和水苏糖的相关系数（R）分别为0.9952、0.9957和0.9877,并且精准度与回收率均较高。使用该方法对低聚糖进行乙酰化反应重现性好、所需样品材料及试剂量少且污染毒害小,能够得到理想的分离、检测和定量分析效果,适用于少量植物组织中低聚糖的定量分析。该方法在食品、医药检测和基础科学研究领域均具有广泛的适用性及参考价值。     

植物组织中低聚糖乙酰化及毛细管气相色谱分析

建立一种用乙酰化衍生处理低聚糖并用毛细管气相色谱-FID进行分析的方法。以1-甲基咪唑为催化剂并以乙酸酐为乙酰化试剂, 同时对植物样品中蔗糖、棉子糖和水苏糖等低聚糖乙酰化产物进行毛细管气相色谱分离和FID检测。确定了低聚糖乙酰化衍生物的毛细管气相色谱分析条件, 并对低聚糖乙酰化反应条件及色谱分离条件进行了优化。结果表明, 在80–1 000 ng·μL^–1范围内线性关系良好, 蔗糖、棉子糖和水苏糖的相关系数(R)分别为0.995 2、0.995 7和0.987 7, 并且精准度与回收率均较高。使用该方法对低聚糖进行乙酰化反应重现性好、所需样品材料及试剂量少且污染毒害小, 能够得到理想的分离、检测和定量分析效果, 适用于少量植物组织中低聚糖的定量分析。该方法在食品、医药检测和基础科学研究领域均具有广泛的适用性及参考价值。     

耐消化道逆环境双歧杆菌菌株A04体外促进生长因子的研究*

研究了4种低聚糖、8种中药及4种食品原料体外对双歧杆菌菌株A04的增殖作用。结果表明,大豆低聚糖的增殖效果最明显(P<0·01),其次是低聚异麦芽糖和低聚果糖,水苏糖则几乎没有增殖效果,而且当改良MRS中低聚糖的浓度达到10%时,对其反而具有一定的抑制作用;8种中药在浓度为2%的时均有增殖效果,以Z₃、Z₁和Z₂效果最显著(P<0·01),但其浓度不同对双歧杆菌菌株A04的增殖影响截然不同;本试验采用的4种食品原料对双歧杆菌均     

大豆低聚糖对牙鲆营养效应的研究：Ⅰ摄食、生长和代谢酶活性

本试验比较研究了不同蛋白源(鱼粉,FM;大豆分离蛋白,SPI)基础饲料中添加大豆低聚糖(SBOS)对牙鲆(Paralichthys olivaceus)摄食率、生长性能和代谢酶活性的影响。分别以FM、SPI作为主要蛋白源,配制了4种等氮等能饲料。其中,饲料FM、SPI分别以FM、SPI作为主要蛋白源;饲料FMO、SPIO分别在饲料FM、SPI基础上添加10%SBOS(水苏糖：2.61%;棉籽糖：0.61%)。试验表明：①饲料中添加SBOS对牙鲆前两周摄食率无显著影响(p > 0.05);而显著降低了整个养殖周期FMO组牙鲆总摄食率,提高了SPIO组牙鲆总摄食率(p < 0.05);②FM基础饲料中添加SBOS对牙鲆特定生长率(SGR)、饲料效率(FER)和蛋白质效率(PER)均无显著影响(p > 0.05),但均呈明显下降趋势;SPI基础饲料中添加SBOS对FER和PER无显著影响,却显著提高了SGR(p < 0.05);③FM基础饲料中添加SBOS提高了牙鲆血浆谷草转氨酶(AST)、谷丙转氨酶(ALT)、碱性磷酸酶(AKP)、乳酸脱氢酶(LDH)和γ-L-谷氨酰转肽酶(γ-GT)的活性,但仅AST差异显著(p < 0.05),而对肝脏AST、ALT、AKP、LDH和γ-GT活性无显著影响(p > 0.05);SPI基础饲料中添加SBOS显著提高了血浆ALT和LDH活性,而对AST、AKP和γ-GT活性无显著影响;同时,SPI基础饲料中添加SBOS显著提高了肝脏AST和ALT活性,而对AKP、LDH和γ-GT活性无显著影响;④饲料中添加SBOS对牙鲆血浆中尿素氮和游离氨基酸浓度均无显著影响。试验结果表明,不同蛋白源基础饲料中添加SBOS表现出不同的生长效应;其中,FM基础饲料中添加SBOS表现出一定的抑制生长效应,而SPI基础饲料中添加SBOS却表现出促生长效应。     

Contents of soluble carbohydrates in yellow lupin seeds maturated at various temperatures

The objective of this paper was to compare the levels of soluble sugars in seeds of yellow lupin cv. Juno matured at different temperatures. The temperature regimes applied were 1). 26 °C for 24 h (high temperature), 2). 24 °C for 12 h and 19 °C for the next 12 h (optimum temperature regime), 3). 26 °C for 16 h and 4 °C for the next 8 h (high-low temperatures). Six soluble carbohydrates (d-galactose, myo-inositol, sucrose, raffinose, stachyose and verbascose) were quantified. Seeds maturing at constant temperature 26 °C accumulated more raffinose (by 100 %) than seeds maturing at optimum temperature regime. Seeds maturing at high temperature accumulated less stachyose and verbascose than those maturing at optimum temperature conditions, the differences being 45 and 24 %, respectively. In seeds maturing at high-low temperature the level of raffinose decreased while the level of stachyose and verbascose increased, compared to those maturing at optimum conditions. The contents of sucrose, d-galactose and myo-inositol in seeds maturing at optimum temperatures was lower than in seeds maturing at both high and high-low temperature regimes. It was shown, that temperature conditions — constant high temperature, or physiologically optimal thermal oscillations (24 °/19 °C) or high-low temperature regime — differently affect the contents of six soluble carbohydrates in maturing seeds of yellow lupin.     

Naturally occurring prebiotic oligosaccharides in poultry feed mixtures

The presence of raffinose series oligosaccharides (RSO) was determined by an enzymic method in three commercially available chicken feed mixtures. All feed mixtures contained RSO at a concentration of 2.1–2.2 %. Soya meal was identified as the exclusive source of RSO. Subsequently, the bifidogenic effect of stachyose (main soya bean RSO) was also assigned on the growth of poultry intestinal bifidobacteria. Bifidobacteria were counted in chicken intestinal tract using cultivation and FISH methods. Four out of 6 bifidobacterial strains tested grew significantly better on stachyose than on glucose. It can be thus concluded that chicken feed mixtures naturally contain prebiotic oligosaccharides in the form of RSO in higher levels (>2 %) compared with the concentration (usually up to 1 %) recommended for artificially added prebiotics. Our results therefore indicate that there is no reason for the supplementation of chicken feed mixtures with prebiotics with bifidogenic properties.     

Nonstructural carbohydrates in dormant and afterripened wild oat caryopses

Nonstructural carbohydrates were determined in both embryo and endosperm of dormant (nongerminating) and afterripened (germinating) intact caryopses of wild oat ( Avena fatua L.). No changes in endosperm starch or soluble sugar were observed at the onset of germination (18 h). No changes in glucose, fructose, sucrose or starch within dormant or afterripened embryos correlated with onset of visual germination. In afterripened embryos, depletion of raffinose (18 h), stachyose (18 h) and galactose (24 h) was correlated with germination. In contrast, raffinose-family oligosaccharide levels in dormant embryos remained constant for 7 days following imbibition. Germination of isolated dormant embryos on 88 m M galactose-containing media was accompanied by decreased endogenous levels of raffinose and stachyose. Isolated embryos from dormant caryopses incorporated ¹⁴C from ¹⁴C-fructose into both raffinose and stachyose during 24 h of imbibition. In contrast, no ¹⁴C incorporation into stachyose was observed in embryos from afterripened caryopses. No ¹⁴C incorporation into raffinose was observed at 18 and 24 h. When in vitro activities of α galactosidase were measured, no temporal differences between dormant or afterripened caryopses were detected in either embryo or endosperm tissue. Although the mechanism associated with differences in utilization of raffinose and stachyose is yet unidentified, alterations in raffinose-family oligosaccharide metabolism in the embryo appear to be a unique prerequisite for afterripening-induced germination.     

大豆低聚糖对牙鲆营养效应的研究：Ⅱ 消化率和消化生理

本试验比较研究了不同蛋白源(鱼粉,FM;大豆分离蛋白,SPI)基础饲料中添加大豆低聚糖(SBOS)对牙鲆(Paralichthys olivaceus)消化道(胃、幽门盲囊、前肠、中肠和后肠)和肝脏消化酶活性、饲料表观消化率和消化道(胃、小肠)组织结构的影响。分别以FM、SPI作为主要蛋白源,配制了4种等氮等能饲料。其中,饲料FM、SPI分别以FM、SPI作为主要蛋白源;饲料FMO、SPIO分别在饲料FM、SPI基础上添加10%SBOS(水苏糖：2.61%;棉籽糖：0.61%)。试验表明：①饲料中添加SBOS普遍降低了牙鲆消化道和肝脏蛋白酶活性,但差异均不显著(p > 0.05)。FM基础饲料中添加SBOS显著降低了肝脏脂肪酶活性(p < 0.05),而对消化道脂肪酶活性无显著影响;SPI基础饲料中添加SBOS显著提高了前肠脂肪酶活性,降低了胃和中肠脂肪酶的活性,而对幽门盲囊、后肠和肝脏脂肪酶活性均无显著影响。FM基础饲料中添加SBOS显著降低了中肠淀粉酶活性,提高了胃淀粉酶活性(p < 0.05),而对幽门盲囊、前肠、后肠和肝脏淀粉酶活性无显著影响;SPI基础饲料中添加SBOS显著提高了胃和前肠淀粉酶活性,降低了中肠脂肪酶活性(p < 0.05),而对幽门盲囊、后肠和肝脏淀粉酶活性无显著影响;②FM基础饲料中添加SBOS显著降低了饲料干物质表观消化率(p < 0.05),而对粗蛋白、粗脂肪和能量表观消化率均无显著影响(p > 0.05)。SPI基础饲料中添加SBOS对干物质、粗蛋白、粗脂肪和能量表观消化率均无显著影响;③饲料中添加SBOS对牙鲆胃和小肠组织结构均无明显负面影响。试验结果表明,饲料中添加SBOS对牙鲆消化道和肝脏消化酶活性、饲料表观消化率和消化道组织结构均没有表现出明显的负面效应;大豆蛋白源中含有的SBOS不是影响牙鲆对其利用率差的主要因素。     

α-Galactosidase from sweet chestnut seeds

The major sugars of fresh seeds of Castanea sativa were shown to be raffinose, stachyose and sucrose. Drying seeds at 25° for 14 weeks increased the ratio raffinose: stachyose from 1.1 to 3.5, reduced sucrose content by ca 50 % and decreased total extractable α-galactosidase. The enzyme activity was resolved into two peaks, a high MW form I (apparent MW215 000) and a low MW form II (apparent MW 53 000). The latter form was predominant in the extract of fresh seeds whereas the former was the main form in the 14-week dried seeds. An increase in the amount of enzyme I was also observed when a buffered extract (pH 5.5) of fresh seeds was stored at 4°. Enzymes I and II had pH optima of 4.5 and 6, respectively. Both enzymes hydrolysed p-nitrophenyl α-d-galactoside at a much greater rate than the natural substrates raffinose, stachyose, locust bean gum and carob gum. However, enzyme I showed preference for stachyose as compared to raffinose; the opposite order was observed for enzyme II.     

Characterization and biotechnological application of an acid alpha-galactosidase from Tachigali multijuga Benth. seeds

Tachigali multijuga Benth. seeds were found to contain protein (364 mg g(-1)dwt), lipids (24 mg g(-1)dwt), ash (35 mg g(-1)dwt), and carbohydrates (577 mg g(-1)dwt). Sucrose, raffinose, and stachyose concentrations were 8.3, 3.0, and 11.6 mg g(-1)dwt, respectively. alpha-Galactosidase activity increased during seed germination and reached a maximum level at 108 h after seed imbibition. The alpha-galactosidase purified from germinating seeds had an M(r) of 38,000 and maximal activity at pH 5.0-5.5 and 50 degrees C. The enzyme was stable at 35 degrees C and 40 degrees C, but lost 79% of its activity after 30 min at 50 degrees C. The activation energy (E(a)) values for p-nitrophenyl-alpha-d-galactopyranoside (pNPGal) and raffinose were 13.86 and 4.75 kcal mol(-1), respectively. The K(m) values for pNPGal, melibiose, raffinose, and stachyose were 0.45, 5.37, 39.62 and 48.80 mM, respectively. The enzyme was sensitive to inhibition by HgCl(2), SDS, AgNO(3), CuSO(4), and melibiose. d-Galactose was a competitive inhibitor (K(i)=2.74 mM). In addition to its ability to hydrolyze raffinose and stachyose, the enzyme also hydrolyzed galactomannan.     

Carbohydrate Changes during Maturation of Cucumber Fruit : Implications for Sugar Metabolism and Transport

Changes in the carbohydrate profiles in the mesocarp, endocarp, and seeds of maturing cucumber (Cucumis sativus, L.) fruit were analyzed. Fruit maturity was measured by a decrease in endocarp pH, which was found to correlate with a loss in peel chlorophyll and an increase in citric acid content. Concentrations of glucose and fructose (8.6-10.3 milligrams per gram fresh weight, respectively) were found to be higher than the concentration of sucrose (0.3 milligrams per gram fresh weight) in both mesocarp and endocarp tissue. Neither raffinose nor stachyose were found in these tissues. The levels of glucose and fructose in seeds decreased during development, but sucrose, raffinose, and stachyose accumulated during the late stages of maturation. Both raffinose and stachyose were found in the seeds of six lines of Cucumis sativus L. This accumulation of raffinose saccharides coincided with an increase in galactinol synthase activity in the seeds. Funiculi from maturing fruit were found to be high in sucrose concentration (4.8 milligrams per gram fresh weight) but devoid of both raffinose and stachyose. The results indicated that sucrose is the transport sugar from the peduncle to seed, and that raffinose saccharide accumulation in the seed is the result of in situ biosynthesis and not from direct vascular transport of these oligosaccharides into the seeds.     

Galactinol synthase activity and soluble sugars in developing seeds of four soybean genotypes

Galactinol synthase (UDP-galactose:inositol galactosyltransferase) is the first unique enzyme in the biosynthetic pathway of raffinose saccharides. Its role as a regulator of carbon partitioning between sucrose and raffinose saccharides in developing soybean (Glycine max L. Merrill) seeds was examined. Galactinol synthase activity and concentrations of sucrose, stachyose, and raffinose were compared during seed development between two genotypes that were high and two genotypes that were low in mature seed raffinose saccharide concentration. In all genotypes, sucrose concentration increased as seed development progressed, but in both low raffinose saccharide genotypes, greater increases in sucrose concentration were observed late in seed development. Sucrose to stachyose ratios in mature seeds were 2.3-fold greater in low raffinose saccharide genotypes than in the high raffinose saccharide genotypes. During seed development, higher levels of galactinol synthase activity were observed in the high raffinose saccharide genotypes than in the low raffinose saccharide genotypes. A common linear relationship for all four soybean genotypes was shown to exist between galactinol formed estimated from galactinol synthase activity data and the concentration of galactose present in raffinose saccharides. Results of this study implied that galactinol synthase is an important regulator of carbon partitioning between sucrose and raffinose saccharides in developing soybean seeds.     

Fermentation of stachyose and raffinose by hind-gut bacteria of the weanling pig

Two pigs were weaned at 28 d of age, and one pig each was placed on a corn-soy (CS) or a corn-soy diet containing 40% lactose (CSL). After 28 d a fecal sample was taken from each pig. The fecal bacterial community was fractionated and used as a source of inoculum to determine if high levels of lactose added to CS diets would modify the structure of the hind-gut microbial community and the in vitro breakdown of stachyose (soy molasses served at the source of stachyose) and raffinose. Bacterial growth rate tended to the higher with the CSL diet. Higher growth rates for bacteria from the CSL-fed pig were supported by the higher acetate: propionate production when compared to the CS diet. All the stachyose and raffinose disappeared during the 48 h fermentation. To our knowledge, this is the first report that stachyose or raffinose are completely fermented by the hind-gut bacteria of the weanling pig, and that this process can be affected by the addition of high levels of lactose to the diet.