根瘤菌胞外多糖的结构与信号功能

共生固氮菌如根瘤菌属(Ｒｈｉｚｏｂｉｕｍ)、中华根瘤菌属(Ｓｉｎｏｒｈｉｚｏｂｉｕｍ)等,能在豆科作物的根部形成独特的根瘤结构,并在根瘤内将氮气固定为氨供宿主利用。根瘤菌与宿主植物的共生体系的形成是有种属特异性的,也就是说一种根瘤菌只能侵染为数不多的几种植物。根瘤形成因子(ｎｏｄｆａｃｔｏｒ,ＮＦ)对这种特异性起了重要的决定作用。ＮＦ是在植物类黄酮的诱导下由根瘤菌合成的一种脂酰基甲壳寡糖,不同根瘤菌的ＮＦ在结构上是不同的。除ＮＦ之外,根瘤菌胞外多糖(ｅｘｏｐｏｌｙｓａｃｃｈａｒｉｄｅｓ,ＥＰＳ)是对宿主植物特…     

紫云英根瘤菌胞外多糖合成基因exol的序列分析

对互补紫云英根瘤菌ＥｘｏＮｄｖＦｉｘ变种的２．６ｋｂＤＮＡ片段进行序列测定,分析结果表明,该片段内存在的一个完整的阅读框架（ＯＲＦ）ｅｘｏｌ。ｅｘｏｌ编码了３４０个氨基酸,为细胞质蛋白。Ｅｘｏｌ蛋白序列与苜蓿根瘤菌的糖基转移酶ＥｘｏＵ有较强的同源性,利用启动子检测质粒,分析ｅｘｏｌ基因的表达,发现在ｅｘｏｌ基因上有两个启动子活性区段,Ｐｅｘｏｌａ位于基因前端,Ｐｅｘｏ１ｂ位于基因中间,Ｐｅｘｏｌａ     

紫云英根瘤菌107菌株酸性胞外多糖的分离纯化及结构分析

用化学沉淀法和柱层析法,分离纯化了紫云英根瘤菌野生型１０７菌株,胞外多糖合成缺陷型变种ＮＡ０２及其回复子ＮＡ０２（Ｒ‘－１１）产生的酸性胞外多糖（ＥＰＳ）。结构组成分析表明：菌株１０７与ＮＡ０２（Ｒ’－１１）产生的ＥＰＳ糖组分均由葡萄糖、半乳糖、核糖和葡萄糖醛酸构成,而变种ＮＡ０２产生的ＥＰＳ只含葡萄糖、半乳糖和葡萄糖醛酸,与野生型显著不同。３个菌株的ＥＰＳ均有乙酰基取代,而无其它形式的取代基。Ｅ     

根瘤菌N613胞外多糖发酵条件及抗肿瘤作用研究

用摇瓶正交试验研究了根瘤菌Rhizobium sp.N613合成根瘤菌胞外多糖(rhizobium exopolysaccharide,REPS)的最佳培养基配方和最适培养条件。采用10L自动控制罐进行了分批发酵试验,获得了合成REPS的发酵动力学参数。经40h的补料分批发酵与代谢调控,REPS产量达到11.31g/L。此外,抗肿瘤实验表明,当REPS剂量为5mg/kg时,其抑瘤率可达53.40%。结果表明,REPS的发酵周期短,产量高,成本低,且具有良好的免疫活性与增稠性,有着极高的开发应用价值。     

紫云英根瘤菌Exo^—变种的生理遗传及胞外多糖组分分析

从质粒ｐＪＢ１１－Ｂ６的８．５ｋｂ野生型ＤＮＡ片段中克隆到５．８ｋｂ和２．６ｋｂ的ＤＮＡ片段,其中５．８ｋｂ的片段互补紫云英根瘤菌１０７菌株的胞外多糖（ＥＰＳ）缺陷型变种ＮＡ０６和ＮＡ１２,２,２．６ｋｂ的片段只能互补变种ＮＡ１２。回接实验和电镜切片显示,这种个变种的根瘤与野生型显著不同,无固氮活性,根瘤内基本不含类菌体;它们的Ｅｘｏ＾＋回复子的根瘤与野生型根瘤相似,多数细胞胞含有类菌体,但仍无固     

紫云英根瘤菌胞外多糖缺陷型变种的分离及遗传分析

经转座子Tn5诱变,获得20株紫云英根瘤菌菌株109胞外多糖(Exopolysaccharide,EPS)缺陷型(Eps-deficient,Exo-)变种。这些变种仍都是原养型,在含不同的碳底物或不同浓度碳底物的固体培养基上,菌落型态均无改变。与亲本菌相比,变种的EPS产量明显降低。所有变种未能在其宿主植物紫云英上结瘤。利用载体质粒pMN2,经Exo-变种与大肠杆菌受体菌交配,通过选择Tn5卡那霉素抗性标记的转移,构建成带有Tn5及菌株109多糖基因DNA片段的R－prime质粒。R-prime质粒能稳定地存在于菌株109中。大部分变种的Exc-表型能被R-prime质粒互补,但R-prime质粒未能使这些Exo-变种恢复有效结瘤的共生能力。根据互补结果,20株Exo-变种分为6种不同的互补群,其中5种互补群的多糖位点在遗传上是连锁的。     

紫云英根瘤菌胞外多糖缺失变种及其结瘤性状的变化

根据紫云英根瘤菌在寄主豆科植物紫云英上的结瘤能力,经转座子Tn5诱变获得的18株Exo~-变种可分为4种结瘤类群(A-D):A类变种诱导植物产生小的瘤状突起,不具固氮能力;B类变种形成无效根瘤;C类变种产生固氮效率降低的根瘤;D类变种丧失了结瘤能力。电镜分析显示:无效瘤和瘤状突起中不存在类菌体区,根瘤细胞均为不含细菌的空细胞,侵染线不能穿透到根瘤细胞中。说明紫云英根瘤菌胞外多糖很可能参与有效根瘤的形成。     

不产生胞外多糖的假单胞菌突变菌株E_（16）

在适宜培养条件下,Ｐｓｅｕｄｏｍｏｎａｓｓｐ３１２６０能将木糖转化为酸性胞外多糖（ＥＰＳ）,用甲基磺酸乙醋（ＥＭＳ）诱变处理　Ｐｓｅｕｄｏｍｏｎａｓｓｐ３１２６０得到一株完全不产生胞外多糖的突变菌株Ｅ＿（１６）。     

光强和氮源对念珠藻胞外多糖分泌的影响

胞外多糖（EPS）是结皮蓝藻形成生物结皮的胶结剂,为了理解常球状存在的丝状蓝藻Nostoc胶结沙粒的机理,探讨了光强40、80 E/（m2s）和氮源（气态氮,硝态氮）对结皮优势种Nostoc sp.分泌EPS（包括荚膜多糖CPS和释放多糖RPS）的影响规律及其内在机理。结果发现:Nostoc sp.在气态氮和硝态氮下都有相似的快速生长,但其分泌的RPS、CPS及EPS量,在硝态氮下均随光强的增加而增加,在气态氮下却与光强没有关系。相关代谢研究发现,在硝态氮下细胞内有更高含量的可溶性糖和蔗糖。进一步的相关分析发现,在两种氮源下,蔗糖量与RPS量或CPS量间的显著正相关都只发生在80 E/（m2s）下,在气态氮中,两光强下的胞内总糖量都与CPS量显著负相关。以上结果说明,Nostoc sp.在氮源利用和光强适应方面都有明显优势,它即使在快速生长的对数期,也可同时分泌相当量的EPS,这使其在球状藻殖段形成之前胶结沙粒成为可能。由此可推知,Nostoc sp.在贫瘠沙土表面的最初生长过程中,其胞外的EPS均来自胞内的固碳产物,在高光强下,蔗糖很可能是其EPS合成的原料。       

EXTRACELLULAR POLYSACCHARIDES FROM ANKISTRODESMUS DENSUS (CHLOROPHYCEAE)

The dissolved extracellular compounds produced by the Chlorococcalean Ankistrodesmus densus Kors. were investigated for their molecular masses and chemical composition. Gel permeation chromatography showed apparent relative molecular masses of 2 × 10⁶ and 10⁴, respectively, for two distinct fractions, termed A and B. The higher molecular weight fraction (A) contained mainly fucose and 3- O -methylgalactose, whereas galactose, glucose, mannose, and rhamnose were present in smaller amounts. Methylation analyses showed that the main structural features are fucose as a highly branched part of polymer A with substitutions in both position 2 and position 4, the substitutions on one of those positions being primarily terminal 3- O -methylgalactose and the other one involved as the linkage of the main chain of the polymer. Because of the presence of both fucose and 3- O -methylgalactose, this polymer is highly hydrophobic. In fraction B, mannose represented more than 60% of the carbohydrate material present, whereas the remaining part contained rhamnose, fucose, xylose, and glucose in almost equal amounts. 3- O -methylgalactose and galactose were present as minor elements. Fraction B is basically a mannose-containing polymer in which the mannose units are either 1→4 or 1→2 linked. Traces of glucuronic acid and protein were present in both fractions; neither sulfate nor phosphate was detected.     

几种担子菌胞外漆酶同工酶谱的研究

从猴头(Hericium erinaceus)、亚侧耳(Hohenbuehelia serotina)、毛柄金钱菌(Co-llybia velutipes)、多脂鳞伞(Pholiota adiposa)、香菇(Lentinus edodes)、黑木耳(Auriculariaauricula)、草菇(Volvaria volpacea)二孢蘑菇(Agaricus bisporus)和紫孢侧耳(Pleurotussapidus)、侧耳(Pleurotus ostreatus)风尾菇(P.sajor-caju)、金顶侧耳(P.citrinopileatus)、鲍鱼菇(P.cystidiosus)、佛罗里达侧耳(P.florida)、爪哇漏斗状侧耳(P.sajor-caju var.javanicus)商品化菌种的培养滤液里所得到的胞外漆酶(LC),采用聚丙烯酰胺凝胶等电聚焦盘状电泳方法,分析比较了它们的胞外漆酶同工酶。经多批次比较试验,获得了侧耳属七个种和八个不同属间各自恒定的漆酶同工酶谱。在这些酶谱中,分别包含了 A 组(pH3—5)和 B 组(pH5—8)的谱带。而且,所有菌种间漆酶带的等电点值(pI)各有异同。没有测定菌种间各漆酶带含量差别。讨论了漆酶同工酶谱用于分类研究的可能性。     

STRUCTURE OF EXTRACELLULAR POLYSACCHARIDES PRODUCED BY A SOIL CRYPTOMONAS SP. (CRYPTOPHYCEAE)1

The Hindak strain of a Cryptomonas species (Cryptophyceae) produces extracellular polysaccharides. Because there is no information on the structure of these compounds in the Cryptophyceae we conducted structural studies. Gas–liquid chromatographic analyses showed that the polysaccharide is composed of fucose, rhamnose, xylose, mannose, glucose, galactose, galacturonic acid, glucuronic acid, and traces of 3-O-methyl galactose. The polysaccharide was separated into two subtractions by ion-exchange chromatography. Fraction A consisted mainly of 1,3-linked galactose units and 1,4-linked galacturonic acid. Unlike fraction B, fraction A did not have xylose, 3-O-methyl galactose, or glucuronic acid. Also, its degree of branching was low compared to that of fraction B. Only traces of sulfate were present infraction A, but fraction B was 10–15% sulfated. Protein was approximately 1% in both fractions. These polysaccharides appear to be a novel type of polymer in algae.     

多粘芽孢杆菌204产生的高粘性多糖性质的研究

多粘芽孢杆菌(Bacillus polymyxa)204能在萄葡糖培养基中,产生具有高粘性的胞外多糖,发酵48—72h产率为12g/L。在浓度为0.6％,1.0％和1.5％时,它的粘度分别为327,3715和44650厘泊、该菌产生的胞外多糖是一种酸性多糖,它由葡萄糖、甘露糖和半乳糖组成;其克分子比为5:5:2;糖醛酸含量为5.5％;其中1％水溶液具有加热熔化、冷却后凝固的类琼脂性质。本文还讨论了浓度、pH、加热温度和时间,以及添加盐类对比多糖的影响。毒性试验证明,此多糖是无毒的,LD50>10g/kg。     

兜唇石斛多糖的研究

从兜唇石斛（Ｄｅｎｄｒｏｂｉｕｍａｐｈｙｌｌｕｍ）的茎中提取到３种多糖：ＡＰ－１,Ａｐ一２,Ａｐ一３,玻璃纤维纸电泳、超速离心和凝胶层析证明：三者都为单一均匀成分。３种多糖的特性粘数（［ｎ］）分别为４２６．９６,４１６．９６,２３４．０３;分子量（Ｍｗ）分别为８６３００,６１５００,４３１００;旋光度（）分别为－３０．０００,－３３．０００,１７．０００。经化学方法和光谱分析证明：ＡＰ为Ｂ（１→４）连接的、含Ｏ一乙酰基的、吡喃型的直链Ｄ一葡萄甘露聚糖。动物体内试验表明：ＡＰ有免疫增强作用。     

一种测定艰难梭菌荚膜多糖分子量的简便方法

本文采用两种规格的Ｓｅｐｈａｒｏｓｅ４Ｂ凝胶层析柱,测定了艰难梭菌荚膜多糖的分子量,并建立了一种较为简便、敏感的测定方法,对进一步研究英膜多糖的生物学特性提供了有利条件。     

COMPARATIVE STUDIES ON THE CELL WALLS OF SEXUAL AND ASEXUAL BANGIA ATROPURPUREA (RHODOPHYTA). I. HISTOCHEMISTRY OF POLYSACCHARIDES1

A comparative histochemical study of the nature and distribution of acidic and neutral cell wall polysaccharides was conducted on marine sexual and asexual filaments of the red alga Bangia atropurpurea (Roth) C. Ag. Outer and inner walls of this species were clearly partitioned according to staining and transmission electron microscopic characteristics. Neutral polysaccharides were detected in the outer coating (cuticle) but were absent from outer and inner walls of all filaments. Acidic polysaccharides were noted in the outer wall material but not in the inner wall layers of any filaments at any developmental stages. The major staining component of vegetative regions of sexual material and all regions of asexual filaments was a highly sulfated polymer. During sexual reproduction only there was a generalized change in the nature of the acidic component, characterized by a decrease in intensity of staining for sulfates in both male and female filaments and the appearance, in female filaments only, of polysaccharides which presumably were carboxylated. Spermatia attached to both male and female filaments in regions where sexual differentiation was initiated and where changes in the outer wall components commenced.