壳寡糖的制备及其在医学和农业生产中的应用

壳寡糖为一种由2~10个氨基葡萄糖经β-1,4糖苷键连接而成的寡糖聚合物,可通过化学水解或酶降解几丁质或壳聚糖获得,在医学及农业生产等各个领域具有广泛的研究和应用价值。壳寡糖天然无毒,分子量相对较低,水溶性好,易于吸收,且具有良好的生物相容性。此外,壳寡糖也表现出了良好的生物学活性,包括抗肿瘤、抗炎、免疫调节、抗菌、改善糖脂代谢紊乱、保护神经损伤等。对壳寡糖的制备和表征、生物学功能及应用进展进行了综述,并对壳寡糖产业目前存在的问题及未来的研究方向进行了讨论,以期为壳寡糖的深度开发提供依据。     

不同取代度N-邻苯二甲酰壳聚糖抗氧化活性的研究

低聚壳聚糖与邻苯二甲酸酐酰化得到三种取代度不同的N-邻苯二甲酸酐酰低聚壳聚糖NPCOSA、NPCOSB和NPCOSC,取代度分别为0.330、.55和0.65。通过红外光谱对其结构进行表征。并考察了其抗氧化性能。结果表明:COS的抗氧化性能最强;随着取代度的增加,N-邻苯二甲酰低聚壳聚糖对超氧阴离子的清除能力逐渐升高;而对DPPH的清除能力以及还原能力呈逐渐下降趋势;对羟基自由基的清除顺序大小依次为NPCOSB>NPCOSA>NPCOSC,即NPCOSB清除羟基自由基的的能力最佳。     

细菌脂多糖及其寡糖链结构分析技术研究进展

脂多糖是大多数革兰氏阴性细菌细胞壁的主要成分,能诱发宿主细胞固有免疫反应,在细菌的识别、黏附、转移、致病等过程中发挥非常重要的作用.其结构组成与细菌的血清型和致病能力息息相关,因而对其结构进行精准分析,有助于研究其结构与生物效应间的关系,便于鉴别菌种和研发相关的抗生素及疫苗.由于脂多糖具有两亲性、多电荷且结构复杂等特点,为研究分析带来很多难题.本文全面归纳了细菌脂多糖分析技术的相关研究成果,阐述脂多糖及其寡糖链的提取、分离纯化及鉴定方法.     

甘露寡糖分离纯化研究进展*

甘露寡糖具有润肠通便、降血脂、抗结肠炎症、增强免疫及调节肠道菌群等生理功能,在食品药品等领域具有广阔的应用前景。水解法制得的甘露寡糖含有单糖、未分解聚糖、不同聚合度寡糖及盐离子等杂质,还需要进一步分离纯化。综述了近年来国内外甘露寡糖的主要纯化方法包括柱层析法、膜分离法、乙醇沉淀法和微生物发酵法等。总结了各种方法的原理、应用范围和应用实例,并对不同方法的优缺点进行了分析。     

甘露寡糖分离纯化研究进展*

甘露寡糖具有润肠通便、降血脂、抗结肠炎症、增强免疫及调节肠道菌群等生理功能,在食品药品等领域具有广阔的应用前景。水解法制得的甘露寡糖含有单糖、未分解聚糖、不同聚合度寡糖及盐离子等杂质,还需要进一步分离纯化。综述了近年来国内外甘露寡糖的主要纯化方法包括柱层析法、膜分离法、乙醇沉淀法和微生物发酵法等。总结了各种方法的原理、应用范围和应用实例,并对不同方法的优缺点进行了分析。     

N-酰化低聚壳聚糖衍生物还原能力的研究

对低聚壳聚糖进行N-酰化改性,制得取代度相同的N-马来酰低聚壳聚糖(NMCOS),N-琥珀酰低聚壳聚糖(NSCOS),N-邻苯二甲酰低聚壳聚糖(NPCOS),其中NMCOS1、NSCOS1、NPCOS1的取代度均为0.25;NMCOS2、NSCOS2、NPCOS2的取代度均为0.49。考察了6种N-酰化低聚壳聚糖衍生物的还原能力。结果表明:当取代度相同时,N-邻苯二甲酰低聚壳聚糖的还原能力最强,其次是N-马来酰低聚壳聚糖,N-琥珀酰低聚壳聚糖的还原能力最差。这可能是由取代基的性质不同所致。     

Oligosaccharide mapping of proteoglycan-bound and xyloside-initiated dermatan sulfate from fibroblasts

The copolymeric structure of dermatan sulfate chains synthesized by skin fibroblasts has been examined. Chains initiated onto exogeneousp-nitrophenyl--D-xylopyranoside or attached to protein in a large proteoglycan, PG-L, and two small proteoglycans, PG-S1 and PG-S2, have been compared by using high resolution electrophoresis and gel chromatography of oligosaccharides generated by specific enzymatic or chemical degradations. The results confirm that chains attached to PG-L are glucuronate-rich, whereas novel findings indicate that chains attached to either of the two PG-S variants yield closely similar oligosaccharide maps, have approximately equal glucuronate and iduronate content and contain over 90% 4-sulfated disaccharide repeating units. Dermatan sulfate chains built onto xyloside at concentrations of 50 µm and below have a copolymeric structure similar to that of chains from the two PG-S variants. These findings indicate that the polymer-modifying machinery can generate chains with extended iduronate-containing repeats also when the xylose primer is not linked to core protein.     

真菌寡糖诱导植物抗性活性成分的分离纯化

寡聚糖作为一种信号分子 ,在调节植物的生长、发育以及植物在不同环境中生存能力等方面起着非常重要的作用[1] .许多特定结构的寡糖被证明具有诱导植物抗性的作用 .对具有诱导抗性作用的葡聚寡糖结构分析表明 ,其最小活性寡糖单位是由 7个葡葡糖残基组成的 β 葡聚糖苷 ,在一个     

高活性壳聚糖酶制剂的制备及其对壳聚糖降解作用的研究

对系列壳聚糖酶高产菌株的产酶性能及产酶发酵液的壳聚糖酶活性进行了比较,从中筛选出一株优良芽孢杆菌菌株,其产酶发酵液的壳聚糖酶活力高达5000U/mL(以单位时间内底物壳聚糖的减少量确定酶活力)。利用此粗制壳聚糖酶制剂对壳聚糖进行酶解产糖的研究表明:壳聚糖的转化率及壳寡糖的产率在适合的酶解条件下,短时间内即可接近100%。     

牛蒡寡糖对黄瓜植株生理生化特性的影响

用不同浓度的牛蒡寡糖(BOS)溶液喷施黄瓜幼苗植株,研究牛蒡寡糖对黄瓜正常生长的生理生化特性的影响.结果表明,与对照组相比,施用1、3、5、8和10 g?L-1牛蒡寡糖处理的黄瓜叶片其叶绿素含量、游离脯氨酸含量、可溶性糖含量、可溶性蛋白含量和PSⅡ的光化学效率均有不同程度的升高,而细胞膜相对透性和MDA的含量均有所降低,其中5 g?L-1是最适处理浓度.因此,牛蒡寡糖能够促进植物的生长发育并提高其抗逆性.     

低聚壳聚糖美拉德衍生物抑菌性能研究

本文研究了基于与葡萄糖、麦芽糖和木糖进行美拉德反应的低聚壳聚糖衍生物的抑菌性.测定低聚壳聚糖及其衍生物对大肠杆菌和金黄色葡萄球菌的抑制效果.结果显示:壳聚糖及其衍生物对金黄色葡萄球菌的抑制作用强于对大肠杆菌的抑制作用,且随着浓度增加,对两种菌的抑菌效果增强.大多数壳聚糖衍生物的抑菌效果优于壳聚糖本身,其中CG 1∶1 8 h(低聚壳聚糖的氨基与葡萄糖的羰基的物质量比为1∶1,反应8h)的抑菌效果最好,CM 1∶3 8 h(低聚壳聚糖的氨基与麦芽糖的羰基的物质量比为1∶3,反应8 h)抑菌性最差,这可能与参加反应的还原糖种类、反应物比例以及反应时间相关.     

红花菜豆凝集素与莫寡糖部分分子间的聚合

红花菜豆凝集素与莫寡糖部分分子间的聚合施炜星,孙册（中国科学院上海生物化学研究所,上海,２０００３１）关键词红花菜豆凝集素;寡糖;分子间聚合凝集素是一类与糖结合并凝集细胞或沉淀含糖大分子的蛋白质或糖蛋白。凝集素均由亚基组成,通常一个亚基具有一个结合糖...     

唾液酸苷酶的催化机理及水解与合成寡糖进展

唾液酸苷酶(EC.3.2.1.18)是一类重要的糖苷水解酶,在动物和微生物中广泛存在.该类酶催化寡糖或糖缀合物上非还原末端唾液酸水解,具有重要的生物学功能,如参与溶酶体降解代谢物、癌症发生、微生物致病等多种生理和病理过程.除了水解活性外,有的唾液酸苷酶还具有转糖基活性,能够以唾液酸单糖或糖苷为糖基供体,催化唾液酸转移到受体分子上,一步合成寡糖和糖苷化合物.这种合成活性对于唾液酸相关糖链的大量获得具有重要意义,有利于推动该类寡糖的基础研究及其在食品和医药中的应用.本文综述了唾液酸苷酶的结构和催化机理、生理功能、转糖基作用及其在寡糖合成中的应用.     

Crystal structures and inhibitor binding properties of plant class V chitinases: the cycad enzyme exhibits unique structural and functional features

A class V (glycoside hydrolase family 18) chitinase from the cycad Cycas revoluta (CrChiA) is a plant chitinase that has been reported to possess efficient transglycosylation (TG) activity. We solved the crystal structure of CrChiA, and compared it with those of class V chitinases from Nicotiana tabacum (NtChiV) and Arabidopsis thaliana (AtChiC), which do not efficiently catalyze the TG reaction. All three chitinases had a similar (α/β)₈ barrel fold with an (α + β) insertion domain. In the acceptor binding site (+1, +2 and +3) of CrChiA, the Trp168 side chain was found to stack face‐to‐face with the +3 sugar. However, this interaction was not found in the identical regions of NtChiV and AtChiC. In the DxDxE motif, which is essential for catalysis, the carboxyl group of the middle Asp (Asp117) was always oriented toward the catalytic acid Glu119 in CrChiA, whereas the corresponding Asp in NtChiV and AtChiC was oriented toward the first Asp. These structural features of CrChiA appear to be responsible for the efficient TG activity. When binding of the inhibitor allosamidin was evaluated using isothermal titration calorimetry, the changes in binding free energy of the three chitinases were found to be similar to each other, i.e. between ?9.5 and ?9.8 kcal mol^?1. However, solvation and conformational entropy changes in CrChiA were markedly different from those in NtChiV and AtChiC, but similar to those of chitinase A from Serratia marcescens (SmChiA), which also exhibits significant TG activity. These results provide insight into the molecular mechanism underlying the TG reaction and the molecular evolution from bacterial chitinases to plant class V chitinases.     

Induction of Volatile Organic Compounds of Lycopersicon esculentum Mill. and Its Resistance to Botrytis cinerea Pers. by Burdock Oligosaccharide

In the present study, we investigated the induction of volatile organic compounds （VOCs） of Lycoperslcon esculentum Mill. and its resistance to Botrytis cinerea Pers. by burdock oligosaccharlde. The disease severity of L. esculentum was evaluated 48 h after treatment with 0.6% burdock oligosaccharlde, followed by inoculation with a spore suspension of B. cinerea. The formation of O2＇, the activity of lipoxygenases （LOX）, peroxidase （POD）, cataiase （CAT）, and superoxide dismutase （SOD）, and the quantity and quallty of changes In VOCs were determined a period of time after treatment with 0.6% burdock ollgosaccharide. The results demonstrated that the disease index in treated plants was decreased by 42.5% compared with control 96 h after Inoculation. The production of O2＇ reached a maximum 6 h after treatment （1.36-fold compared with control）. There was an increase in LOX, POD, CAT and SOD activity in response to burdock oligosaccharide treatment and the enzymes showed different trends in the time-course of induction. At 120 h after treatment, （E）-2-hexenal was increased by 92% compared with control, whereas methyl salicylate showed a gradual Increase with induction period. Previous results had demonstrated that chitosan elicitor enhanced the production VOCs of L. esculentum and decreased plant susceptibility towards B. clnerea. Together, these findings suggest that increasing the production of VOCs in response to burrdock oligosaccharide may be an important mechanism for L. esculentumin its defense against pathogens, in addition, burrdock oligosaccharlde may act as a potent elicitor of resistance to disease in L. esculentum.     

人胎盘纤连蛋白N糖链结构研究

分离纯化了人胎盘纤连蛋白（Ｆｎ）,经ＳＤＳ－ＰＡＧＥ鉴定为一条带,纯化Ｆｎ仍保持其搞原性,得率为３８．７％。根据植物凝集素识别专一糖链结构的原理,应用斑点印迹法,亲和层析法和Ｗｅｓｔｅｒｎ转移电泳研究糖链结构,结果证实：１．人胎盘Ｆｎ分子中含有复杂型Ｎ糖链（包括二天线和大于二天线的结构）以及高甘露糖型和／或杂合型Ｎ糖链;复杂型Ｎ糖链中含有平分型ＧｌｃＮＡｃ,糖链末端也可连有唾液酸;２．胰糜蛋白酶水解而获得的明胶结合片段（４４ｋＤ）含有二天线和多天线复杂型糖链,也可接有平分型ＧｌｃＮＡｃ;３．肝素结合片段（３０ｋＤ）以及明胶、肝素均不结合的Ｆｎ片段不含有多天线复杂型Ｎ糖链。     

寡糖对SPF大鼠肠道微生物多样性影响的研究

目的：研究寡糖对SPF大鼠肠道微生物的影响作用。方法：从饲养30d的SPE大鼠盲肠内容物中直接提取总DNA,PCR扩增获得16S rDNA片段,进行DCGE分析。结果：通过DGGE条带数目及亮度的变化,反映出饲喂寡糖能提高和改善肠道内有益菌的数量和组成。结论：饲喂不同水平的寡糖对大鼠肠道微生物菌群影响较大。     

cDNA文库的构建策略及其应用

cDNA文库在基因分离和克隆中具有重要的作用。从cDNA文库中能筛选出所需要的目的基因,并直接用于该目的基因的表达。cDNA文库是发现新基因和研究基因功能的基础工具。随着分子生物学技术的发展。cDNA文库构建方法有了很大改进和提高,就cDNA文库的构建方法及其应用进行综述。     

人胚肺成纤维细胞株纤连蛋白结构域N─糖链结构研究

本文应用明胶、肝素亲和层析二步法首先纯化了人胚肺成纤维细胞培养液的纤连蛋白（Ｆｉｂｒｏｎｅｏｔｈｅ,Ｆｎ）,经ＳＤＳ－ＰＡＧＥ鉴定为一条带,然后用胰糜蛋白酶消化纯化的Ｆｎ所获得的酶解波,经分离分别得到明胶结合片段和肝素结合片段,再应用凝集素－ＨＲＰ染色的Ｗｅｓｔｅｎ转移电泳法研究糖链结构,结果证实：１．Ｆｎ中明胶结合片段（４４ｋｄ）中含有二天线和多天线复杂型糖链,并接有平分型ｇｌｃＮＡｃ糖基、核心力Ｆｕｃ。２肝素结合片段（３０ｋｄ）只含有二天线复杂型糖链,不含平分型ＧｌｃＮＡｃ糖基及核心Ｆｕｃ．     

小刺猴头菌发酵浸膏寡糖对荷瘤小鼠免疫功能的影响

通过建立S180荷瘤小鼠试验动物模型,研究小刺猴头菌发酵浸膏寡糖对S180荷瘤小鼠免疫功能的影响。选择灌胃的方法将不同组别小刺猴头菌发酵浸膏寡糖喂服到荷瘤小鼠体内,观察肿瘤生长情况,对其进行免疫功能的测定。结果表明:小刺猴头菌发酵浸膏寡糖可增加荷瘤小鼠免疫器官脾脏和胸腺质量,提高荷瘤小鼠血清的IL-2和TNF-α含量,抑制肿瘤生长,提高机体免疫功能。