海藻多糖的单糖组成对体外抗氧化活性的影响

比较广西北部湾石莼(Ulva lactuca L.)、海带(Laminaria japonica)、裙带菜(Undaria pinnatifida Surin-gar)、紫菜(Porphyra)的单糖组成及抗氧化活性的差异,揭示多糖结构与其体外抗氧化活性的关系。利用PMP柱前衍生化HPLC分析海藻多糖的单糖组成,采用羟自由基清除试验、超氧阴离子自由基清除试验及DPPH自由基清除试验指征其体外抗氧化活性,结果表明,4种海藻多糖的单糖组成在主成分空间分布离散,石莼及紫菜主要由葡萄糖组成,海带主要由甘露糖组成,裙带菜则主要由半乳糖组成;其体外抗氧化活性存在显著差异,裙带菜多糖对DPPH的清除能力(半抑制浓度IC₅₀值为0. 56±0. 02 mg/mL)显著高于其他3种海藻多糖;石莼、裙带菜与海带对羟自由基均有较强的清除活性,而紫菜多糖对羟自由基的清除能力较差(IC₅₀值为26. 59±0. 98mg/mL);石莼与裙带菜对超氧阴离子的清除活性较强,显著高于海带与紫菜,其中石莼显著高于裙带菜,IC₅₀值分别为1. 61±0. 17、2. 73±0. 06 mg/mL。相关性分析及冗余分析结果表明,对抗氧化活性影响较为显著的因子为葡萄糖(Glc)、核糖(Rib)、木糖(Xyl)(P <0. 01)。     

Isolation and characterization of Aspergillus sp. for the production of extracellular polysaccharides by response surface methodology

In this study, Aspergillus sp. was isolated for the production of extracellular polysaccharide. The process parameters were initially optimized by traditional methods. The cheap substrate, wheat bran was used for the production of extracellular polysaccharide in solid state fermentation. Supplementation of (1%, w/w) maltose, gelatin enhanced EPS production (5.36?mg/g). The salts such as, Cu²⁺ (4.9?mg/g), Ca²⁺ (3.5?mg/g), Zn²⁺ (2.9?mg/g), Mn²⁺ (3.4?mg/g) and Mg²⁺ (1.8?mg/g) stimulated EPS production. In two level full factorial experimental designs, the EPS yield varied from 3.18 to 11.65?mg/g wheat bran substrate with various combinations of the components supplemented with wheat bran substrate. Among these selected factors in central composite design, maltose significantly influenced on extracellular polysaccharide production.     

枸杞多糖抑制SMMC-7721肝癌细胞的VEGF表达、迁移与侵袭

目的研究枸杞多糖(lycium barbarum polysaccharide,LBP)对SMMC-7721肝癌细胞迁移、侵袭影响的机制。方法运用MTT法检肝癌细胞SMMC-7721的增殖率,Transwell检测细胞的迁移力和侵袭力,应用血管内皮生长因子(vascular endothelial growth factor,VEGF)小分子干扰RNA沉默VEGF表达,转染pcDNA 3.1-VEGF过表达VEGF,Western blot和qRT-PCR检测VEGF、MMP-2和MMP-9表达。结果 LBP(100、200、400μg/ml)处理可抑制SMMC-7721细胞的增殖、迁移和侵袭,并抑制MMP-2、MMP-9和VEGF表达;沉默VEGF可降低SMMC-7721细胞迁移、侵袭相关蛋白MMP-2和MMP-9水平,过表达VEGF可逆转LBP对SMMC-7721细胞迁移和侵袭相关蛋白MMP-2和MMP-9水平的抑制作用。结论枸杞多糖可抑制肝癌细胞的迁移和侵袭,其机制可能与直接抑制VEGF有关。     

6A型肺炎球菌结合物分子大小对小鼠免疫原性的影响

目的 比较不同分子大小的6A型肺炎球菌(serotype 6A Streptococcus Pneumoniae )结合物和佐剂吸附在小鼠体内免疫原性的影响。方法 通过乙酸水解降低6A型荚膜多糖的相对分子质量制备成水解物,水解物经1-氰基-4-二甲胺基吡啶四氟硼酸盐(CDAP)活化并与破伤风类毒素己二酸酰肼衍生物TT AH 结合,制备成结合物。用Sepharose 4 Fast Flow 纯化结合物,并根据化学检测结果将结合物分为 K D 0.0~0.2、 K D 0.2~0.4、 K D 0.4~0.6等3个组分,每个组分分别以磷酸铝佐剂吸附,将吸附前后的各个组分按照每针次每只小鼠0.2 μg分别免疫小鼠,并采用ELISA检测结合物在小鼠体内的抗体水平。结果 3种不同相对分子质量吸附前后的结合物在小鼠体内均能产生较高水平的抗体,各组2、3针之间具有明显的加强效应。在吸附组和未吸附组中,3种不同分子大小的结合物在小鼠体内产生抗体水平无明显差异。各组分佐剂吸附后的结合物血清抗体滴度高于未吸附组,但这种差异无统计学意义( P > 0.05)。结论 结合物的分子大小对小鼠体内抗体水平的产生没有明显影响;磷酸铝佐剂吸附对于不同分子大小的结合物在小鼠体内的免疫原性有一定的增强效应,但这种增强效应差异无统计学意义。     

不同酸水解酪蛋白对W135群和Y群脑膜炎奈瑟菌荚膜多糖产量的影响

目的 探索不同酸水解酪蛋白对W135群与Y群脑膜炎奈瑟菌(脑膜炎球菌)荚膜多糖产量的影响。方法 分别以NaCl质量分数为37%和14%的酸水解酪蛋白作为有机氮源配制改良半综合高盐培养基和低盐培养基,利用全自动细菌发酵罐分别在两种培养基里培养W135群和Y群脑膜炎球菌,比较这两种菌株在两种培养基中的生长时间、收获液菌密度( A 600 nm 值)、收获液去菌体后与去复合多糖后上清中的荚膜多糖含量,以及纯化后的精糖产量;比较高盐培养基收获液及其2倍稀释液中不同终含量的十六烷基三甲基溴化铵(cetyltrimethylammonium bromide, CTAB)溶液对多糖沉淀效果的影响。结果 W135群与Y群脑膜炎球菌在两种培养基中的培养时间差异无统计学意义( P >0.05),但高盐培养基收获液的菌密度、收获液去菌体后上清液中的多糖含量均高于低盐培养基收获液,差异有统计学意义( P < 0.05),而高盐培养基收获液纯化后的精糖产量低于低盐培养基,差异有统计学意义( P <0.05)。高盐培养液2倍稀释后,在CTAB终体积分数为0.04%时,W135群与Y群脑膜炎球菌多糖全部沉淀,而未稀释高盐培养液即使CTAB终体积分数提高到0.14%,依然也不能完全沉淀 W135群与Y群脑膜炎球菌多糖。结论 不同酸水解酪蛋白对W135群和Y群脑膜炎球菌的生长密度和荚膜多糖产量有影响,用CTAB溶液沉淀荚膜多糖时需控制收获液盐含量。     

Purification of a conjugated polysaccharide vaccine using tangential flow diafiltration

Conjugated vaccines prepared from the capsular polysaccharide of Streptococcus pneumoniae can provide immunization against invasive pneumococcal disease, meningitis, and otitis media. One of the critical steps in the production of these vaccines is the removal of free (unreacted) polysaccharides from the protein-polysaccharide conjugate. Experimental studies were performed to evaluate the effects of membrane pore size, filtrate flux, and solution conditions on the transmission of both the conjugate and free polysaccharide through different ultrafiltration membranes. Conjugate purification was done using diafiltration performed in a linearly-scalable tangential flow filtration cassette. More than 98% of the free polysaccharide was removed within a 5-diavolume diafiltration process, which is a significant improvement over previously reported results for purification of similar conjugated vaccines. These results clearly demonstrate the opportunities for using ultrafiltration/diafiltration for the final purification of conjugated vaccine products.     

Ca-transport in sea urchin unfertilized eggs: Regulation by endogenous sulfated polysaccharides and K

Previous data from our laboratory showed that the reticulum of the sea cucumber smooth muscle body wall retains both a sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA) and a sulfated polysaccharide. In this invertebrate, the transport of Ca²⁺ by the SERCA is naturally inhibited by these endogenous sulfated polysaccharides. The inhibition is reverted by K⁺ leading to an enhancement of the Ca²⁺ transport rate. We now show that vesicles derived from the endoplasmic reticulum of unfertilized eggs from the sea urchin Arbacia lixula retain a SERCA that is able to transport Ca²⁺ at the expense of ATP hydrolysis. As described for the sea cucumber SERCA isoform, the enzyme from the sea urchin is activated by K⁺ but not by Li⁺ and is inhibited by thapsigargin, a specific inhibitor of SERCA. A new sulfated polysaccharide was identified in the sea urchin eggs reticulum composed mainly by galactose, glucose, hexosamine and manose. After extraction and purification, this sulfated polysaccharide was able to inhibit the mammal SERCA isoform found in rabbit skeletal muscle and the inhibition is reversed by K⁺. These data suggest that the regulation of the SERCA pump by K⁺ and sulfated polysaccharides is not restricted to few marine invertebrates but is widespread.     

真菌多糖抗肿瘤及免疫调节作用研究进展

真菌多糖被认为是目前最有开发前途的保健食品和药品的新资源之一,对真菌多糖的生物活性研究也是保健食品功能因子中最为炽热的研究热点。本文综述了近年来国内外真菌多糖抗肿瘤及免疫调节作用的研究进展。     

发酵茯苓菌丝体和天然茯苓多糖的研究

本文对发酵茯苓菌丝体和天然茯苓中多糖的提取分离进行了研究,并分别测定了二者总多糖的提取率及总糖的平均含量。发酵茯苓菌丝体和天然茯苓中总多糖的提取率分别为24.47%和82.93%;发酵茯苓菌丝体和天然茯苓总糖的平均含量分别为45.17%和87.24%。     

锁阳多糖提取工艺的研究

以浸膏得率和浸膏中多糖质量分数综合评价,利用L9(34)正交实验从水提和醇沉两方面优选锁阳中水溶性多糖最佳提取工艺条件,为锁阳活性多糖的开发提供科学依据和实验基础。最佳水提工艺条件为:m(锁阳):v(水)=1:12,回流提取0.5 h,提取2次;最佳醇沉工艺条件为:在m(生药):v(水提液)=1 g:2 mL溶液中加入乙醇,使φ(乙醇)=70%,静置6 h。