Succinylation of cyclodextrin glycosyltransferase from Thermoanaerobacter sp. 501 enhances its transferase activity using starch as donor

A simple modification procedure, the succinylation of amino groups, was suitable to increase the transferase (disproportionation) activity of cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacter sp. 501 using different linear oligosaccharides as acceptors. On the contrary, the synthesis of cyclodextrins (CDs), the coupling of CDs with oligosaccharides, and the hydrolysis of starch decreased after chemical modification. The degree of succinylation of amino groups (45%) was accurately determined by MALDI-TOF mass spectrometry. The formation of CDs under industrial conditions was analyzed for native and succinylated CGTases, showing similar selectivity to alpha-, beta-, gamma-CD. The acceptor reaction with D-glucose using soluble starch as glucosyl donor was studied at 60 degrees C and pH 5.5. Malto-oligosaccharides (MOS) production was notably higher using the semisynthetic enzyme at different ratios (w/w) starch:D-glucose. Thus, more than 90% of the initial starch was converted into MOS (G2-G7) in 48 h employing a ratio donor:acceptor 1:2 (w/w).     

Structure of the glycerol phosphate-containing O-specific polysaccharide and serological studies on the lipopolysaccharides of Citrobacter werkmanii PCM 1548 and PCM 1549 (serogroup O14)

The O-specific polysaccharide was obtained by mild acid hydrolysis of the lipopolysaccharide of Citrobacter werkmanii PCM 1548 and PCM 1549 (serogroup O14) and found to contain D-glucose, D-glucosamine and glycerol-1-phosphate in molar ratios 2 : 2 : 1. Based on methylation analysis and 1H and 13C nuclear magnetic resonance spectroscopy data, it was established that the O-specific polysaccharides from both strains have the identical branched tetrasaccharide repeating unit with 3,6-disubstituted GlcNAc, followed by 2,4-disubstituted Glc residues carrying at the branching points lateral residues of Glc and GlcNAc at positions 6 and 2, respectively. Glycerol-1-phosphate is linked to position 6 of the chain Glc. All sugars have a beta configuration, except for the side-chain Glc, which is alpha. Serological studies revealed a close relatedness of the lipopolysaccharides of C. werkmanii PCM 1548 and PCM 1549, both belonging to serogroup O14. In immunoblotting, anti-C. werkmanii PCM 1548 serum showed no cross-reactivity with the O-polysaccharide bands of the lipopolysaccharides of Citrobacter youngae PCM 1550 (serogroup O16) and Hafnia alvei PCM 1207, also containing a lateral glycerol phosphate residue.     

Structural studies on N-acetylmannosaminuronic acid-containing and glucuronic acid-containing teichuronic acids in the cell wall of Bacillus megaterium AHU 1375

Structural studies were carried out on two kinds of teichuronic acid-glycopeptide complexes (designated as TU-GP-I and TU-GP-II) isolated from lysozyme digest of N-acetylated cell walls of Bacillus megaterium AHU 1375 by ion-exchange chromatography and gel chromatography. TU-GP-I, accounting for about 25% of the cell walls, contained N-acetylmannosaminuronic acid, N-acetylglucosamine, glucose, galactose, glycerol, and phosphorus in an approximate molar ratio of 1:1:2:1:0.5:0.5, together with small amounts of glycopeptide components. TU-GP-II, accounting for about 9% of the cell walls, contained glucuronic acid, glucose, and fucose in a molar ratio of about 2:1.5:1, together with small amounts of glycopeptide components. The results of analyses involving Smith degradation, chromium oxidation, methylation, acetolysis, and H-NMR measurement led to the conclusion that the polysaccharide chain of TU-GP-I comprised repeating units,----6) Glc(alpha 1----3)-ManNAcUA(beta 1----4)[Gal(alpha 1----3)][Glc(beta 1----6)]GlcNAc(beta 1----. About half of the repeating units were substituted by glycerophosphoryl residues at C-6 of the beta-glucosyl residues linked to the N-acetylglucosamine residues. By means of a similar procedure, the polysaccharide chain of TU-GP-II was shown to comprise repeating units,----4)GlcUA(alpha 1----3)GlcUA(alpha 1----3)Glc(alpha 1----3)Fuc(alpha 1----, of which about half were substituted by alpha-glucosyl residues at C-3 of the 4-substituted glucuronosyl residues.     

The simultaneous determination of the products of estrogen 2- and 4-hydroxylase action; the use of high-performance liquid chromatography with electrochemical detection

Both trace-labeled and high-specific activity ¹²⁵I-labeled derivatives of hexadecapeptide gastrin (G) were prepared by reaction with the iodinated form of the imidoester, methyl p-hydroxybenzimidate. Reaction conditions for preparation of trace-labeled iodinated imidoester gastrin (IIE-G) were: excess imidoester to G (IIE:G, 20:1), pH 9.2, and a reaction time of 24 h. Following purification by gel filtration and silica gel chromatography, an IIE-G component was isolated which appeared homogeneous on thin-layer chromatography and retained the same biological and immunological properties as unmodified G. Somewhat different conditions were necessary to prepare high specific activity iodinated imidoester gastrin (IIE¹-G). These included reducing the volume (20 μl) and pH (7.5) at which the imidoester was iodinated and adjusting the concentrations of reactants to the same molar amounts as 5 mCi of carrier-free ¹²⁵I. Sufficient amounts of IIE¹-G were obtained by reversing the ratio of G and IIE¹ and reacting with a G excess (GIIE¹, 10:1). The purified IIE¹-G had a specific activity exceeding 1500 μCi/nmol and was used to establish a specific and sensitive radioimmunoassay for gastrin.     

刺五加果水溶性多糖的研究——AS-2的分离纯化与结构探讨

从刺五加果中抽提出水溶性粗多糖。经酸性乙醇分级及反复冻融得到多糖AS-2。AS-2经Sepharose CL-4B柱层析为单一对称峰,经醋酸纤维素膜电泳为一条带,冻融后高速离心无沉淀可证明其为均一级分。G.C分析表明,AS-2由Ara、Xyl、Rha、Gal、Glc组成,其单糖摩尔比为1.6:1.2:1.8:1.0:3.6。AS-2的分子量约为78kD,比旋光度[α]_D~(25)=+17°,特性粘度[η]=0.068。红外光谱分析含β型糖苷键。部分酸水解、酶解、高碘酸酸化、Smith降解、完全甲基化、G.C,G.C-M.S的分析结果表明,以β(1→3)Glc及β(1→4)Glc构成分子的主链。Glc的C_3上带有分支,约每4个己糖残基带有1个侧链。侧链上,Rha多以1→4苷键相连,部分残基C_2上有分支。Gal存在(1→6)及(1→3)连接方式,多数Glc以(1→6)苷键连结,少数Glc出现在分子非还原末端。位于分子末端的还有Ara与Xyl。     

Synthesis of Malto-Oligosaccharides Via the Acceptor Reaction Catalyzed by Cyclodextrin Glycosyltransferases

The disproportionation activity (intermolecular transglycosylation) of cyclomaltodextrin glycosyltransferases (CGTases) from Thermoanaerobacter sp. and Bacillus circulans strain 251 was studied. Using soluble starch as donor, the CGTase from Thermoanaerobacter sp. showed the highest transglycosylation activity with all the malto-oligosaccharides tested as acceptors. At ratios of starch: D-glucose from 2:1 to 1:2 (w/w), the formation of cyclodextrins was completely inhibited, and a homologous series of malto-oligosaccharides (G_n) was produced. The conversion of starch into acceptor products was in the range of 63-79% in 48 h. The degree of polymerisation of malto-oligosaccharides formed could be modulated by the ratio of starch: D-glucose provided; at a ratio of 1:2 (w/w), the reaction was quite selective for the formation of G2-G3.     

Starch biosynthesis: the primer nonreducing-end mechanism versus the nonprimer reducing-end two-site insertion mechanism

Two mechanisms are recognized for polysaccharide chain elongation: (a) the nonreducing-end, primer-dependent mechanism and (b) the reducing-end, two-site insertion mechanism. We recently demonstrated the latter mechanism for starch biosynthesis by pulsing starch granules with ADP-[14C]Glc and chasing with ADPGlc for eight varieties of starch granules. Others have reported the addition of glucose from ADPGlc to the nonreducing ends of maltose, maltotriose, and maltopentaose and a branched maltopentasaccharide. It was concluded that starch chains are biosynthesized by the addition of glucose to the nonreducing ends of maltodextrin primers. In this study, we reinvestigated the maltodextrin reactions by reacting three kinds of starch granules from maize, wheat, and rice with ADP-[14C]Glc in the absence and presence of maltose (G2), maltotriose (G3), and maltodextrin (d.p.12) and found that they inhibited starch biosynthesis rather than stimulating it, as would be expected for primers. The major product in the presence of G2 was G3 with decreasing amounts of G4-G9 and the major products in the presence of G3 was G4 and G5, with decreasing amounts of G6-G9. It was concluded that maltodextrins are acceptors rather than primers. This was confirmed by pulsing the starch granules with ADP-[14C]Glc and chasing with G2, G3, and G6, which gave release of 14C-label from the pulsed granules in the absence of ADPGlc, further demonstrating that maltodextrins are acceptors that inhibit starch biosynthesis by releasing glucose from starch synthase, rather than acting as primers and stimulating biosynthesis.     

白魔芋和花魔芋葡甘露聚糖研究

从白魔芋和花魔芋的块茎中分离纯化出两种魔芋葡甘露聚糖(Konjac Glucomannan,简称KGM),分别名为白魔芋葡甘露聚糖(aKGM)和花魔芋葡甘露聚糖(rKGM)。通过超离心、玻璃纤维纸电泳和凝胶过沪证明aKGM和rKGM都是均一的多糖。这两种多糖都由甘露糖(M)和葡萄糖(G)组成,其克分子比G/M:aKGM为1:1.69,rKGM为1:1.60,分子量前者为8.09×10~5,后者为7.37×10~5。酶水解实验和红外光谱分析说明aKGM和rKGM都是由甘露糖和葡萄糖以β-1,4-糖苷键连接的杂多糖;有O-乙酰基的特征吸收峰,说明在某些糖残基上可能有乙酰基团。     

Global Shapes of F-actin Depolymerization-competent Minimal Gelsolins: INSIGHT INTO THE ROLE OF g2-g3 LINKER IN pH/Ca2+ INSENSITIVITY OF THE FIRST HALF*

Because of its ability to rapidly depolymerize F-actin, plasma gelsolin has emerged as a therapeutic molecule in different disease conditions. High amounts of exogenous gelsolin are, however, required to treat animal models of different diseases. Knowing that the F-actin depolymerizing property of gelsolin resides in its N terminus, we made several truncated versions of plasma gelsolin. The smaller versions, particularly the one composed of the first 28–161 residues, depolymerized the F-actin much faster than the native gelsolin and other truncates at the same molar ratios. Although G1-G3 loses its dependence on Ca²⁺ or low pH for the actin depolymerization function, interestingly, G1-G2 and its smaller versions were found to regain this requirement. Small angle x-ray scattering-based shape reconstructions revealed that G1-G3 adopts an open shape in both the presence and the absence of Ca²⁺ as well as low pH, whereas G1-G2 and residues 28–161 prefer collapsed states in Ca²⁺-free conditions at pH 8. The mutations in the g2-g3 linker resulted in the calcium sensitivity of the mutant G1-G3 for F-actin depolymerization activity, although the F-actin-binding sites remained exposed in the mutant G1-G3 as well as in the smaller truncates even in the Ca²⁺-free conditions at pH 8. Furthermore, unlike wild type G1-G3, calcium-sensitive mutants of G1-G3 acquired closed shapes in the absence of free calcium, implying a role of g2-g3 linker in determining the open F-actin depolymerizing-competent shape of G1-G3 in this condition. We demonstrate that the mobility of the G1 domain, essential for F-actin depolymerization, is indirectly regulated by the gelsolin-like sequence of g2-g3 linker.     

Characterization of a novel linkage unit between ribitol teichoic acid and peptidoglycan in Listeria monocytogenes cell walls

The structure of the linkage unit between ribitol teichoic acid and peptidoglycan in the cell walls of Listeria monocytogenes EGD was studied. A teichoic-acid--glycopeptide preparation isolated from lysozyme digests of the cell walls of this strain contained mannosamine, glycerol, glucose and muramic acid 6-phosphate in an approximate molar ratio of 1:1:2:1, together with large amounts of glucosamine and other components of teichoic acid and glycopeptides. A teichoic-acid-linked sugar preparation, obtained by heating the cell walls at pH 2.5, also contained glucosamine, mannosamine, glycerol and glucose in an approximate molar ratio of 25:1:1:2. Part of the glucosamine residues were shown to be involved in the linkage unit. Thus, on mild alkaline hydrolysis, the teichoic-acid-linked sugar preparation gave a disaccharide characterized as N-acetylmannosaminyl(beta 1----4)-N-acetylglucosamine [ManNAc(beta 1----4)GlcNAc] in addition to the ribitol teichoic acid moiety, whereas the teichoic-acid - glycopeptide was separated into disaccharide-linked glycopeptide and the ribitol teichoic acid moiety by the same procedure. Furthermore, Smith degradation of the cell walls gave a characteristic fragment, EtO2-P-Glc(beta 1----3)Glc(beta 1----1/3)Gro-P-ManNAc(beta 1----4)GlcNAc (where EtO2 = 1,2-ethylenediol and Gro = glycerol). The results lead to the conclusion that in the cell walls of this organism, the ribitol teichoic acid chain is linked to peptidoglycan through a novel linkage unit, Glc(beta 1----3)Glc(beta 1----1/3)Gro-P-(3/4)ManNAc-(beta 1----4)GlcNAc.     

Evidence for non-uniform distribution of D-glucose within human red cells during net exit and counterflow

The kinetic parameters of net exit of D-glucose from human red blood cells have been measured after the cells were loaded to 18 mM, 75 mM and 120 mM at 2 degrees C and 75 mM and 120 mM at 20 degrees C. Reducing the temperature, or raising the loading concentration raises the apparent Km for net exit. Deoxygenation also reduces the Km for D-glucose exit from red blood cells loaded initially to 120 mM at 20 degrees C from 32.9 +/- 2.3 mM (13) with oxygenated blood to 20.5 +/- 1.3 mM (17) (P less than 0.01). Deoxygenation increases the ratio Vmax/Km from 5.29 +/- 0.26 min-1 (13) for oxygenated blood to 7.13 +/- 0.29 min-1 (17) for deoxygenated blood (P less than 0.001). The counterflow of D-glucose from solutions containing 1 mM 14C-labelled D-glucose was measured at 2 degrees C and 20 degrees C. Reduction in temperature, reduced the maximal level to which labelled D-glucose was accumulated and altered the course of equilibration of the specific activity of intracellular D-glucose from a single exponential to a more complex form. Raising the internal concentration from 18 mM to 90 mM at 2 degrees C also alters the course of equilibration of labelled D-glucose within the cell to a complex form. The apparent asymmetry of the transport system may be estimated from the intracellular concentrations of labelled and unlabelled sugar at the turning point of the counterflow transient. The estimates of asymmetry obtained from this approach indicate that there is no significant asymmetry at 20 degrees C and at 2 degrees C asymmetry is between 3 and 6. This is at least 20-fold less than predicted from the kinetic parameter asymmetries for net exit and entry. None of the above results fit a kinetic scheme in which the asymmetry of the transport system is controlled by intrinsic differences in the kinetic parameters at the inner and outer membrane surface. These results are consistent with a model for sugar transport in which movement between sugar within bound and free intracellular compartments can become the rate-limiting step in controlling net movement into, or out of the cell.     

Characterization and anti-allergic effect of a polysaccharide from the flower buds of Lonicera japonica

A water-soluble polysaccharide (LJP-1), with a molecular weight of 1.8×10(5)Da, was isolated from the flower buds of Lonicera japonica. Gas chromatography (GC) analysis showed that the LJP-1 was mainly composed of d-glucose and a small amount of d-arabinose. On the basis of methylation analysis, LJP-1 had the backbone chain mainly consisting of 1,6-linked Glc and 1,3,6-linked Glc, which was terminated with 1-linked Ara residues at the O-3 position of 1,3,6-linked Glc in a relative molar ratio of 2.9:1:0.9. The anti-allergic effect of LJP-1 was evaluated on allergic contact dermatitis (ACD) induced by picryl chloride (PC) in mouse ear. Similar to prednisolone, orally administrated LJP-1 (20, 40 and 80mg/kg) potently inhibited the PC-induced ACD, leading to substantial reductions in ear thickness, serum level of IgE and histamine, as well as tissue TNF-α. These results demonstrate that treatment with LJP-1 may be effective for preventing the development of PC-induced ACD.     

Metabolism of alpha-D-[1,2-13C]glucose pentaacetate and alpha-D-glucose penta[2-13C]acetate in rat hepatocytes

Hepatocytes from fed rats were incubated for 120 min in the presence of alpha-D-[1,2-13C]glucose pentaacetate (1.7 mM), both D-[1,2-13C]glucose (1.7 mM) and acetate (8.5 mM), alpha-D-glucose penta[2-13C]acetate (1.7 mM), or D-[1,2-13C]glucose (8.3 mM). The amounts of 13C-enriched L-lactate and D-glucose and those of acetate and beta-hydroxybutyrate recovered in the incubation medium were comparable under the first two experimental conditions. The vast majority of D-glucose isotopomers consisted of alpha- and beta-D[1,2-13C]glucose. The less abundant single-labeled isotopomers of D-glucose were equally labeled on each C atom. The output of 13C-labeled L-lactate, mainly L-[2-13C]lactate and L-[3-13C]lactate, was 1 order of magnitude lower than that found in hepatocytes exposed to 8.3 mM D-[1,2-13C]glucose, in which case the total production of the single-labeled species of D-glucose was also increased and that of the C3- or C4-labeled hexose was lower than that of the other 13C-labeled isotopomers. In cells exposed to alpha-D-glucose penta[2-13C]acetate, the large majority of 13C atoms was recovered as [2-13C]acetate and, to a much lesser extent, beta-hydroxybutyrate labeled in position 2 and/or 4. Nevertheless, L-[2-13C]lactate, L-[3-13C]lactate, and single-labeled D-glucose isotopomers were also produced in amounts higher or comparable to those found in cells exposed to alpha-D-[1,2-13C]glucose pentaacetate. However, a modest preferential labelling of the C6-C5-C4 moiety of D-glucose, relative to its C1-C2-C3 moiety, and a lesser isotopic enrichment of the C3 (or C4), relative to that of C1 (or C6) and C2 (or C5), were now observed. These findings indicate that, despite extensive hydrolysis of alpha-D-glucose pentaacetate (1.7 mM) in the hepatocytes, the catabolism of its D-glucose moiety is not more efficient than that of unesterified D-glucose, tested at the same molar concentration (1.7 mM) in the presence of the same molar concentration of unesterified acetate (8.5 mM), and much lower than that found at a physiological concentration of the hexose (8.3 mM). The present results also argue against any significant back-and-forth interconversion of D-glucose 6-phosphate and triose phosphates, under conditions in which sizeable amounts of D-glucose are formed de novo from 13C-enriched Krebs cycle intermediates generated from either D-[1,2-13C]glucose or [2-13C]acetate.     

A complex polysaccharide from the seeds of anthocephalus indicus a. rich

The seeds of Anthocephalus indicus contain a water-soluble polysaccharide composed of D-xylose, D-mannose, and D-glucose in the molar ratios 1:3:5. Methylation analysis afforded 2,3,4-tri-O-methyl-D-xylose, 2,3,6,-tri-O-methyl-D-mannose, 2,3,6-tri-O-methyl-D-glucose, 2,3-di-O-methyl-D-glucose, and 2,3,4,6-tetra-O-methyl-D-glucose in the molar ratios 7:21:12:15:8. Periodate oxidation and methylation data indicated 22.5% and 21.9% of end groups, respectively. The above findings, together with the results of partial hydrolysis with acid, indicate the polysaccharide to consist of a linear chain of (1→4)-linked β-D-mannosyl and β-D-glucosyl residues to which α-D-xylosyl and β-D-glucosyl groups are attached by (1→6)-linkages.     

Cyclodextrins micrometric powders obtained by supercritical fluid processing

Supercritical fluid technology offers the possibility to produce dry powder formulations of biocompatible materials, overcoming the drawbacks of classical micronization processes. In this work, Supercritical Assisted Atomization (SAA) has been used to micronize alpha-cyclodextrin (alpha-CD) and hydroxypropyl-beta-cyclodextrin (HP-beta-CD). Some process parameters, such as precipitation temperature and solute concentration in the liquid solution, have been studied to evaluate their influence on morphology and size of precipitated particles. Cyclodextrins (CDs) micronization has been successful: well-defined spherical microparticles of alpha-CD and HP-beta-CD have been produced. Particle size analysis revealed that sharp distributions have been obtained: 95% of particles have diameters ranging between 0.1 and 5 microm for both CDs. X-ray and DSC analyses have been also performed to investigate CDs modifications induced by SAA processing: amorphous particles have been obtained in both cases, whereas raw alpha-CD was crystalline and raw HP-beta-CD was amorphous.     

Structure of the polysaccharide isolated from the sheath of Sphaerotilus natans

A polysaccharide was isolated from the sheath of a sheathed bacterium, Sphaerotilus natans. The sheath polysaccharide (SPS) was composed of D-glucose and D-(N-acetyl)galactosamine in molar ratios of 1:4. Methylation linkage analysis revealed the presence of the residues of 4-linked glucose, 4-linked (N-acetyl)galactosamine, and 3-linked (N-acetyl)galactosamine in molar ratios of 1:3:1. The oligomer of SPS was prepared with an SPS-specific degrading enzyme from a sheath-degrading bacterium, Paenibacillus koleovorans. The oligomer was derivatized and subjected to fast atom bombardment-mass spectrometry to investigate the monosaccharide sequence of SPS. The structure of SPS was confirmed by nuclear magnetic resonance. The resulting data showed that SPS is a straight-chained basic polysaccharide constructed of a pentasaccharide repeating unit.     

草苁蓉根、茎水溶性多糖BRT的结构特征

本文以长白山区珍贵野生药用植物草苁蓉为研究对象 .草苁蓉又名“不老草” ,具有滋补强壮、益寿延年之功及补肾壮阳、润肠止血之效 ,为国家三级重点保护植物[1] .近年的研究发现 ,草苁蓉醇提物不仅可以清除体内的游离基 ,而且还可以显著增强机体的免疫能力 ,同时对草苁蓉化学成分的研究也在逐步深入[2 ] ,但对于草苁蓉多糖的系统研究尚未见报道 .为了更全面地认识和利用草苁蓉这一珍贵的植物资源 ,同时也为探讨多糖的结构与功能的关系 ,本文对草苁蓉根、茎的水溶性多糖ＢＲＴ组分进行了结构测定方面的研究 .1　材料和方法1 1　材料为本研究…     

Isolation, characterization, and antitumor activities of the cell wall polysaccharides from Elsinoe leucospila.

A cell-wall preparation from the cells of Elsinoe leucospila, which produces elsinan extracellularly when grown on sucrose or glucose-potato extract medium, was fractionated systematically. The heteropolysaccharide that was released by treatment with Actinase E digestion, comprised D-mannose, D-galactose, and D-glucose (molar ratio, 1.5:1.0:0.1). Methylation, mild acid hydrolysis, and 13C-NMR studies suggested that the polysaccharide contains a backbone of alpha-(1----6)-linked D-mannose residues having two kinds of side chains, one attached at the O-4 with single or short beta-(1----6)-linked D-galactofuranosyl residues, and the other attached at O-2 with short side chains, most probably, of alpha-(1----3)-linked D-mannopyranosyl residues. A moderately branched D-glucan fraction, obtained from the cold alkali extract, was fractionated to give an antitumor-active purified beta-(1----3)-glucan having branches of single beta-D-glucosyl groups, one out of eight D-glucose residues being substituted at the O-6.     

Isolation and Characterization of a Xyloglucan from Gobo (Arctium lappa L.)

A xyloglucan was isolated from the 24% KOH extract of gobo (edible burdock, Arctium lappa L.). A methylation analysis and enzymic degradation studies on the polysaccharide showed that gobo-xyloglucan was built up predominantly of repeating-oligosaccharide units of hepta-(Glc: Xyl=4: 3), nona- (Glc: Xyl: Gal: Fuc=4: 3: 1: 1) and deca- (Glc: Xyl: Gal: Fuc=4: 3: 2: 1) saccharides in an approximate molar ratio of 14: 12: 5, which are the typical structural units of dicot xyloglucans.     

Structural studies on the specific type VII pneumococcal polysaccharide

The specific type VII pneumococcal polysaccharide was isolated from the crude capsular material by precipitative and chromatographic methods. It contained D-galactose, D-glucose, L-rhamnose, 2-acetamido-2-deoxy-D-glucose, and 2-acetamido-2-deoxy-D-galactose in the molar ratio of 3.5:2.3:3.0:2.1:1.0. Some of its structural features were revealed by methylation studies, time-lapse hydrolysis, periodate oxidation, and enzymic hydrolysis. The polysaccharide is branched at residues of D-galactose and 2-acetamido-2-deoxy-D-galactose. Non-reducing end groups consisted of D-galactopyranose and 2-acetamido-2-deoxy-D-glucopyranose residues, with the former predominating. Major components of the linear chains were (1→3)-linked L-rhamnose and (1→4)-linked D-glucose; the minor ones were (1→2)-linked L-rhamnose, (1→6)-linked D-galactose, and (1→6)-linked 2-acetamido-2-deoxy-D-glucopyranose. The (1→4)-linked D-glucose components may be present as cellobiose residues. The results are in accord with structural features deduced from the serological cross-reactivity of this polysaccharide.