大豆低聚糖的制备工艺研究

研究从脱脂豆粕中提取大豆低聚糖的制备工艺。经多次实验,确定了浸提、活性炭脱色和离子交换脱盐等过程的优化工艺参数,并对成品糖浆的成分进行了测定。     

酶法制备直链麦芽寡糖

为了获得单一聚合度的麦芽寡糖,以环糊精为原料,利用环葡聚糖水解酶的单点水解开环反应活性,在95℃、pH5.5、30min的条件下,生产出高纯度的直链麦芽寡糖;并以此为基础,提出了一种以环糊精葡萄糖基转移酶和环葡聚糖水解酶先后降解淀粉,生产直链麦芽寡糖的新方法。     

低聚糖和多酚结合偏最小二乘判别分析鉴别油菜蜜产地

采用气相色谱-串联质谱技术测定油菜蜜中6 种低聚糖成分,高效液相色谱-串联质谱技术测定油菜蜜中18 种酚酸物质,并结合偏最小二乘判别分析（partial least square-discrimination analysis,PLS-DA）对来自湖北钟祥市、江苏盐城市、青海刚察县3 个具有显著地理、气候、环境差异的51 个油菜蜜样本进行产地鉴别。方差分析结果显示：3 个产地油菜蜜中松二糖含量具有显著性差异,且青海刚察油菜蜜低聚糖含量相对偏高;油菜蜜的18 种多酚类物质中大多数具有显著的地理差异性且湖北钟祥的油菜蜜中多酚含量相对偏高。多元统计分析结果显示多酚具有显著的地理特征性。油菜蜜中低聚糖和多酚的含量结合PLS-DA产地鉴别的预测精度可达到97%。     

酶解麦麸制备阿魏酸低聚糖的研究

采用木聚糖酶水解麦麸制备阿魏酸低聚糖,以阿魏酸低聚糖的量为指标,研究了酶解温度、pH值、酶浓度、底物浓度、作用时间五个因素对酶解效果的影响.并在此基础上,通过中心组合试验确定最佳酶解条件.最终的优化条件是木聚糖酶浓度3%,底物浓度80 g/L,作用时间12 h,此条件下阿魏酸低聚糖提取量为0.831×10-5 moL/g麦麸.     

In vitro fermentation of cereal dietary fibre carbohydrates by probiotic and intestinal bacteria

A range of probiotic and other intestinal bacteria were examined for their ability to ferment the dietary fibre carbohydrates β‐glucan, xylan, xylo‐oligosaccharides (XOS) and arabinoxylan. β‐Glucan was fermented by Bacteroides spp and Clostridium beijerinckii but was not fermented by lactobacilli, bifidobacteria, enterococci or Escherichia coli. Unsubstituted xylan was not fermented by any of the probiotic bacteria examined. However, many Bifidobacterium species and Lactobacillus brevis were able to grow to high yields using XOS. XOS were also efficiently fermented by some Bacteroides isolates but not by E coli, enterococci, Clostridium difficile, Clostridium perfringens or by the majority of intestinal Lactobacillus species examined. Bifidobacterium longum strains were able to grow well using arabinoxylan as the sole carbon source. These organisms hydrolysed and fermented the arabinosyl residues from arabinoxylan but did not substantially utilise the xylan backbone of the polysaccharide. Arabinoxylan was not fermented by lactobacilli, enterococci, E coli, C perfringens or C difficile and has potential to be an applicable carbohydrate to complement probiotic Bif longum strains in synbiotic combinations. © 2002 Society of Chemical Industry     

Effects of oligosaccharides on phase transition temperatures and rheological characteristics of waxy rice starch dispersion

BACKGROUND: The creation of starch‐based foods incorporated with functional ingredients such as probiotics is of great current interest in the food industry. This study aimed to investigate the effects of prebiotic oligosaccharides on the phase transition temperatures and rheological characteristics of waxy rice starch dispersions. Four oligosaccharides were applied to the rice starch dispersions: chitooligosaccharides, fructooligosaccharides, isomaltooligosaccharides and xylooligosaccharides. RESULTS: The addition of 125 g kg^?1 oligosaccharides elevated the onset and peak temperatures for gelatinisation of 200–400 g kg^?1 waxy rice starch dispersions. The temperature of the storage modulus (G′) for gelatinisation increased markedly on adding fructooligosaccharides to 200–300 g kg^?1 waxy rice starch. For gelatinisation of 300 g kg^?1 rice starch dispersion the effectiveness of the oligosaccharides in changing the above parameters was as follows: chitooligosaccharides > fructooligosaccharides > isomaltooligosaccharides > xylooligosaccharides. Moreover, their effectiveness was dependent on the amylose content, as illustrated by comparing waxy and non‐waxy rice starches (amylose contents 9–256 g kg^?1). Importantly, the logarithmic G′₉₅ change was linearly and negatively correlated with amylose content. CONCLUSION: The results suggest that oligosaccharide‐containing rice starch dispersions may potentially be used for the formulation of oligosaccharide‐containing starchy functional foods owing to the rheological changes of these starch dispersions. Copyright © 2011 Society of Chemical Industry     

海藻糖、海藻胶及寡糖对南美白对虾蛋白质冷冻变性的抑制作用

为探索海藻糖类对于冷冻水产品的抗冻保水效果,以南美白对虾为研究对象,以焦磷酸钠为对照,研究海藻糖、海藻胶及寡糖对南美白对虾蛋白质冷冻变性的抑制效果。结果表明,-18℃冻藏6周后,0.5%、1.0%海藻糖和海藻胶寡糖(500~600 Da)处理可显著降低冷冻虾仁解冻汁液流失(5.00~5.54%),其与焦磷酸钠保水效果无显著性差异(5.02~5.48%,p0.05)。整个冻藏期内,0.5%、1.0%焦磷酸钠、海藻糖和海藻胶寡糖处理虾仁,肌肉L*值稍有变化但并不显著(p0.05),表明该3种物质对冷冻虾仁肌肉明度具有良好保护作用。随着冻藏时间延长,不同处理虾仁肌原纤维蛋白、盐溶性蛋白及Ca2+-ATPase活性均呈逐渐下降趋势,其中海藻糖、海藻胶寡糖对虾仁蛋白质特性的保护效果,显著优于蒸馏水、焦磷酸钠处理(p0.05)。本研究为开发一种安全、高效、适用于冷冻虾仁的无磷保水剂提供参考。     

Transglycosylation Activity of Engineered Bifidobacterium Lacto-N-Biosidase Mutants at Donor Subsites for Lacto-N-Tetraose Synthesis

The health benefits of human milk oligosaccharides (HMOs) make them attractive targets as supplements for infant formula milks. However, HMO synthesis is still challenging and only two HMOs have been marketed. Engineering glycoside hydrolases into transglycosylases may provide biocatalytic routes to the synthesis of complex oligosaccharides. Lacto-N-biosidase from Bifidobacterium bifidum (LnbB) is a GH20 enzyme present in the gut microbiota of breast-fed infants that hydrolyzes lacto-N-tetraose (LNT), the core structure of the most abundant type I HMOs. Here we report a mutational study in the donor subsites of the substrate binding cleft with the aim of reducing hydrolytic activity and conferring transglycosylation activity for the synthesis of LNT from p-nitrophenyl β-lacto-N-bioside and lactose. As compared with the wt enzyme with negligible transglycosylation activity, mutants with residual hydrolase activity within 0.05% to 1.6% of the wild-type enzyme result in transglycosylating enzymes with LNT yields in the range of 10–30%. Mutations of Trp394, located in subsite -1 next to the catalytic residues, have a large impact on the transglycosylation/hydrolysis ratio, with W394F being the best mutant as a biocatalyst producing LNT at 32% yield. It is the first reported transglycosylating LnbB enzyme variant, amenable to further engineering for practical enzymatic synthesis of LNT.     

MALDI Q-TOF CID MS for Diagnostic Ion Screening of Human Milk Oligosaccharide Samples

Human milk oligosaccharides (HMO) represent the bioactive components of human milk, influencing the infant’s gastrointestinal microflora and immune system. Structurally, they represent a highly complex class of analyte, where the main core oligosaccharide structures are built from galactose and N-acetylglucosamine, linked by 1–3 or 1–4 glycosidic linkages and potentially modified with fucose and sialic acid residues. The core structures can be linear or branched. Additional structural complexity in samples can be induced by endogenous exoglycosidase activity or chemical procedures during the sample preparation. Here, we show that using matrix-assisted laser desorption/ionization (MALDI) quadrupole-time-of-flight (Q-TOF) collision-induced dissociation (CID) as a fast screening method, diagnostic structural information about single oligosaccharide components present in a complex mixture can be obtained. According to sequencing data on 14 out of 22 parent ions detected in a single high molecular weight oligosaccharide chromatographic fraction, 20 different oligosaccharide structure types, corresponding to over 30 isomeric oligosaccharide structures and over 100 possible HMO isomers when biosynthetic linkage variations were taken into account, were postulated. For MS/MS data analysis, we used the de novo sequencing approach using diagnostic ion analysis on reduced oligosaccharides by following known biosynthetic rules. Using this approach, de novo characterization has been achieved also for the structures, which could not have been predicted.