海洋寡糖及其衍生物活性的研究进展

海洋寡糖是海洋多糖降解得到的含两个至十个单糖残基的糖类化合物。相较于海洋多糖,海洋寡糖具有水溶性高、生物活性强、易于机体吸收等优点。海洋寡糖有抗肿瘤、抗氧化、降低胆固醇、免疫调节等生理活性,可以广泛应用于食品、医药等领域。本文综述了海洋多糖、海洋多糖酶法制备海洋寡糖和酶的催化机制,介绍了褐藻胶、壳聚糖和黄原胶三种主要的海洋多糖,以及褐藻胶裂解酶的β-消除作用机制、壳聚糖酶的置换机制和黄原胶降解酶的降解作用机制。对海洋多糖酶法制备海洋寡糖的不足开展了讨论和未来的发展趋势进行了展望,以期为海洋寡糖制备及应用提供理论依据。     

琼胶寡糖的生物酶解制备工艺研究

以龙须菜为原料,以琼胶得率为指标,采用正交实验研究了液料比、水提温度、水提时间对琼胶得率的影响,得到最佳水提琼胶工艺条件;进一步以水解度为指标,采用响应曲面法研究了pH、酶解温度、底物浓度对琼胶水解度的影响,得到最优酶解工艺。结果表明,琼胶的最佳水提条件为:液料比281(v/w),水提温度120℃,水提时间90 min,在此条件下,琼胶得率为32.8%。酶解时间2h,加酶量20 U/mL条件下的最优酶解工艺为:pH6.4,酶解温度54℃,底物浓度0.6%(w/v),此时水解度为89.75%。经薄层层析分析,酶解产物为偶数新琼寡糖(DP2、4、6、8),其中主要产物为新琼四糖,为功能性琼胶寡糖的开发应用打下基础。      

魔芋葡甘聚糖及其衍生物肠道益生性的体外发酵评价

目的：评价魔芋葡甘聚糖（konjac glucomannan,KGM）及其衍生物脱乙酰基魔芋葡甘聚糖（D-KGM）、魔芋葡甘露低聚糖（konjac oligosaccharide,KOS）的肠道益生性。方法：分别以1 g/100 mL的KGM、KOS和D-KGM为碳源,通过小鼠盲肠内容物体外厌氧发酵,测定发酵液的pH值、微生物和短链脂肪酸,评价肠道益生效果。结果：与阳性对照组（以葡萄糖作为碳源）相比,KGM和KOS发酵液pH值明显降低,短链脂肪酸（short-chainfatty acids,SCFAs）和乳酸菌、双歧杆菌的数量显著增加,肠道潜在致病菌（大肠杆菌、梭状杆菌、拟杆菌）的增殖不明显,而D-KGM发酵液中pH值、肠道微生物、SCFAs与阴性对照组（不含碳源）均无显著性差异。结论：KOS和KGM具有显著的肠道益生性,而D-KGM没有肠道益生性。     

Invited review: Anti-adhesive properties of bovine oligosaccharides and bovine milk fat globule membrane-associated glycoconjugates against bacterial food enteropathogens

The prevalence of the main raw milk and raw milk–derived dairy product enteropathogens (Campylobacter, Shiga toxin-producing Escherichia coli, Listeria, and Salmonella) is higher than the number of epidemiological cases related to ingesting these foodstuffs. Bovine milk oligosaccharides and milk fat globule membrane (MFGM)-linked glycoconjugates interact with foodborne enteropathogens to inhibit their adhesion to intestinal cells and tissues. This review examines the main mechanisms and strategies used by enteropathogens to adhere to their target, details the anti-adhesive properties of MFGM against enteropathogens and enterotoxins, assesses the integrity of bacteria–MFGM complexes during dairy product manufacture and digestion, and discusses the potential for using these macromolecules and glycoconjugates in foods for public health.     

Application of granulated apple pectin in the treatment of hyperlipoproteinaemia I. Deriving the regression equation to describe the changes

 The dynamics of the impact of highly esterified apple pectin on lipid metabolism by humans was studied. The experiments involved 109 patients (54 women and 55 men) with hyperlipoproteinaemia. The lipid metabolism of 73 healthy persons (35 men and 38 women) and 36 persons (17 men and 19 women) with hyperlipoproteinaemia was investigated as a control. A granulated form of a mixture of pectin and sorbitol in a ratio of 2:1 was used. The results confirmed the favourable effect of apple pectin on the major characteristics of lipid metabolism by humans: the levels of serum cholesterol, high-density lipoprotein-cholesterol and triglycerides. Sorbitol manifests synergism regarding the hypolipidaemic effect of pectin. It was established that the model y=x(A+Bx), where x is time and y the parameter under investigation, best explained the dynamics of the studied characteristics of lipid metabolism under the impact of granulated highly esterified apple pectin. (A and B are experimental constants). Received: 29 March 1996/Revised version: 24 July 1996     

Identification of new acceptor specificities of glycosyltransferase R with the aid of substrate microarrays

Finding opportunities to construct sugar motifs and to transfer them to targets of biological relevance and rapid identification of glycosylation events are important goals for glycobiology and a field of increasing interest. Here we have applied an enzyme microarray screening system for the identification of new acceptor specificities of the glycosyltransferase R (GTFR) from Streptococcus oralis (E.C. 2.4.1.5), which was able to effect the synthesis of sugar motifs in short times and with low amounts of substrate. These observations resulted in the development of a convenient alpha-glycosylation by the non-Leloir glycosyltransferase GTFR, with sucrose as substrate and with different alcohols and amino acid derivatives as acceptors, for the synthesis of glycoethers and glycosylated amino acids not observed with the use of familiar GTFs with high sequence homology.     

Chemical Synthesis of Homogeneous Glycoproteins for the Study of Glycoprotein Quality Control System

The glycoprotein quality control system exists in the endoplasmic reticulum to maintain protein homeostasis and prevent accumulation of aberrant glycoproteins. Folding sensor enzyme uridine diphosphate (UDP) glucose : glycoprotein glucosyltransferase (UGGT) plays an important role in this system through its ability to discriminate immature or misfolded glycoproteins from native ones. UGGT transfers a glucose residue to a glycoprotein containing Man₉GlcNAc₂ (M9; Man=mannose, GlcNAc=N-acetyl-D -glucosamine) N-glycan only when the glycoprotein has not attained a native form. We chemically prepared homogeneous glycoproteins containing M9 N-glycan in the native form as well as in misfolded forms and examined them as substrates of UGGT. Glucose transfer to misfolded glycoproteins was clearly observed by LC-MS, but glycoproteins in the native form were barely glucosylated. Furthermore, we constructed an in vitro glycoprotein folding system in the presence of UGGT and found out that all folding intermediates which appeared during folding were also glucosylated. Through these experiments, we demonstrated the usefulness of chemically synthesized homogeneous glycoproteins as probes to gain insights into the molecular basis of the glycoprotein quality control system.