Studies on Mannan in Wood Pulp

The glucomannan isolated from holocellulose pulp of Akamatsu (Pinus densiflora Sieb. et Zucc.) as its triacetate was methylated and the following methylated sugars were obtained by hydrolysis: the 2,3,4,6-tetra-O-methyl ethers of d-glucose and d-mannose (I part) and the 2,3,6-tri-O-methyl ethers of d-mannose and d-glucose (34–37 parts). Periodate oxidation of the glueomannan showed that 1.00 mole of periodate was consumed per mole of hexose unit and 3 moles of formic acid liberated for every 33 hexose units.     

Studies on the Chemical Structure of Konjac Mannan

Rate-constants of β-d-mannosidic and β-d-glucosidic linkages for acid hydrolysis were determined. And from these results, statistical analysis of acid hydrolysis of the main chain of the mannan was made whereby the yields of the fragmental disaccharicles could be calculated for any degree of hydrolysis. The symmetry of the result of these calculations with those observed experimentally may support the proposed chemical structure of the mannan. Moreoyer, from the result of action of cellulase on the manno-oligosaccharides, the main process of the reaction, “hydrolysis of the mannan by cellulase” was discussed.     

Studies on the Chemical Structure of Konjac Mannan

The electrophoretically homogeneous glucomannan isolated from konjac flour was composed of d-glucose and d-mannose residues in the approximate ratio of 1: 1.6. Controlled acid hydrolysis gave 4-O-β-d-mannopyranosyl-d-mannose, 4-O-β-d-mannopyranosyl-d-glucose_T 4-O-β-d-glucopyranosyl-d-glucose(cellobiose), 4-O-β-d-glucopyranosyl-d-mannose(epicellobiose), O-β-d-mannopyranosyl-(1→4)-O-β-d-mannopyranosyl-(1→4)-d-mannose, O-β-d-glucopyranosyl- (1→4)-O-β-d-mannopyranosyl-(1→4)-d-mannose, O-β-d-mannopyranosyl-(1→4)-O-β-d-glucopy- ranosyl-(1→4)-d-mannose and O-β-d-glucopyranosyl-(1→4)-O-β-d-glucopyranosyl-(1→4)-d-mannose.     

锌酵母中酵母甘露多糖组分的特征和结构

本文研究从锌酵母中分离出的酵母甘露多糖ＸＰ的特征和结构。ＸＰ经全水解和＾１３ＣＮＭＲ谱显示除甘露糖基外,还有少量Ｌ－鼠李糖基和甲氧基。甲基化分析、过碘酸盐氧化、Ｓｍｉｔｈ降解、乙酰解和部分酸水解显示ＸＰ的主链是１→６连接的甘露糖,侧链是１→２连接的甘露糖。＾１Ｈ及＾１３Ｃ ＮＭＲ谱表明所有糖苷键均为α型,结合元素分析ＸＰ基本是酵母甘露多糖和蛋白质以及锌的络合物。     

Anatomical Studies of Chunia Wood

Chunia is a monotype of Hamamelidaceae in China. The wood structure of this genus possesses some primitive properties, such as the inclining end-wall of vessel elements, the scalariform perforation plates, the absence of spiral thickening in the wall, the heterogenous rays, crystal inclusion in Cell, and scanty parenchymatous cells in the axial system. On account of its intimate relation to the genus Mytilaria, therefore, both may be represented as primitive genera in this family.     

Studies on Wood Anatomy of Eucalyptus globulus Under Different Habitats

Wood anatomy of Eucalyptus globulus Labill. under different habitats was comparatively studied under light and scanning electron microscope. According to the eleven major quantitative perimeters of wood characteristics of E. globulus viz: vessel frequency, percentage of solitary vessels, vessel diameter, vessel member length; tracheid length and diameter; fibre tracheid length and diameter; libriform fibre length; ray frequency and ray height, the plants under different habitats have been made a comparison. The results showed that growth ring was indistinct, and there was a positive correlation between the quantitative characters of most wood constituent members were positively correlated with the latitude except for some of vessel frequency decreased. The decrease in effects of annual rainfall on the structure of the wood of E. glotrulus were evaluated.     

Mannan synthase activity in the CSLD family

Cellulose Synthase Like (CSL) proteins are a group of plant glycosyltransferases that are predicted to synthesize β-1,4-linked polysaccharide backbones. CSLC, CSLF and CSLH families have been confirmed to synthesize xyloglucan and mixed linkage β-glucan, while CSLA family proteins have been shown to synthesize mannans. The polysaccharide products of the five remaining CSL families have not been determined. Five CSLD genes have been identified in Arabidopsis thaliana and a role in cell wall biosynthesis has been demonstrated by reverse genetics. We have extended past research by producing a series of double and triple Arabidopsis mutants and gathered evidence that CSLD2, CSLD3 and CSLD5 are involved in mannan synthesis and that their products are necessary for the transition between early developmental stages in Arabidopsis. Moreover, our data revealed a complex interaction between the three glycosyltransferases and brought new evidence regarding the formation of non-cellulosic polysaccharides through multimeric complexes.Key words: mannan, mannose, plant cell wall, glycosyltransferase, cellulose synthase like, CSL, biosynthesis, hemicelluloseThe plant cell wall is mainly composed of polysaccharides, which are often grouped into cellulose, hemicelluloses and pectin. Since the discovery of the first cellulose synthase (CESA) genes in cotton fibers,¹ the synthesis of cellulose has been extensively studied.² In contrast, the glycosyltransferases responsible for synthesizing hemicelluloses and pectin are still largely unidentified.^3,4,5 The CESA genes are members of a superfamily that includes genes with a high sequence similarity with CESA genes and are named Cellulose Synthase Like (CSL).⁶ The CSL genes have themselves been grouped into nine families designated CSLA, -B, -C, -D, -E, -F, -G, -H and -J (Figure 1A).^5,6 Mannan and glucomannan synthase activity has been demonstrated in the CSLA family,^7,8,9 while members of the CSLC family have been implicated in synthesis of the xyloglucan backbone.¹⁰ CSLF and CSLH, which are found only in grasses, are involved in synthesis of mixed linkage glucan.^11,12 The function of the remaining CSL families has not been determined. We have reported our research on the CSLD family in a recent publication.¹³ Of all the CSL families, CSLD possesses the most ancient intron/exon structure and is the most similar to the CESA family.⁶ CSLD genes are found in all sequenced genomes of terrestrial plants including Physcomitrella and Selaginella suggesting a highly conserved function throughout the plant kingdom (Figure 1A). Five genes (CSLD1 to CSLD5) and one apparent pseudogene (CSLD6) have been identified in Arabidopsis thaliana.¹⁴ Bernal et al.^14,15 studied knock-out mutants of the individual genes and presented evidence for a role in cell wall biosynthesis for each Arabidopsis CSLD. To elucidate the activity of the CSLD proteins and obtain further understanding of their biological role, we generated double mutants csld2/csld3, csld2/csld5, csld3/csld5 and the triple mutant csld2/csld3/csld5. Immunochemical, biochemical and complementation assays brought evidence that CSLD5 or CSLD2 in concomitance with CSLD3 act as mannan synthases.Open in a separate windowFigure 1(A) Schematic representation of the CESA superfamily phylogeny. The inset on the right is a detailed phylogenetic tree of CSLDs from Selaginella moellendorffii, Arabidopsis thaliana and Oryza sativa. The figure is modified from Ulvskov and Scheller.⁵ (B) Comparison of csld2, csld3, csld5 with Col-0 20 days after germination. The inflorescences of csld2 and csld3 were similar to Col-0 whereas csld5 had a delayed growth. Scale bar: 1 cm. (C) Col-0 and csld2/csld3/csld5 (triple mutant, TM) 40 days after germination. After 40 days, the triple mutant was barely developed and, as shown in the magnified inset, displayed purple coloration indicating accumulation of anthocyanins, a typical stress response. Scale bar: 2 mm.     

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。     

生物信息大分子结构修饰对其功能的影响——酵母甘露聚糖硫酸酯化衍生物的制备及其对HIV-1细胞生长抑制效应的初步研究

研究由安徽大学ADF实验室以拟规模化环保型循环工艺生产流程分离、提取、纯化制得的生物多糖-酵母甘露聚糖(Yeast Mannan,简称Man),在对其结构、性质和生物活性研究的基础上,进一步对其结构进行硫酸酯化修饰,制得新产品硫酸酯化酵母甘露聚糖(Man-Sulfated),并探索Man、Man-Sulfated对HIV-1病毒细胞生长的抑制作用。研究结果表明:1)Man对宿主细胞C8166无毒性;2)实验用HIV-1病毒细胞对宿主细胞C8166有毒性,TCID50为1.1×10~6Cells/m L;3)用氯磺酸∶吡啶(2∶1)甲酰胺法制得的Man-Sulfated对宿主细胞C8166也无毒性,并具有抑制HIV-1病毒细胞生长的功能,而Man则无此新功能;4)Man-Sulfated具有抑制HIV-1生长的效能,其与硫酸酯化修饰Man结构时使用的硫酸酯化试剂类别、方法及反应条件的不同,而引起对Man结构、构象的修饰变化程度不同密切相关。浓硫酸酯化法对Man结构修饰太强烈,构象变化过大,影响其生物功能。     

果寡糖和甘露寡糖对断奶仔猪免疫机能的影响

研究探讨了果寡糖和甘露寡糖对断奶仔猪免疫机能的影响。实验采用随机区组设计法将48头健康、体重一致的35日龄断奶杜洛克仔猪分为4组,每组3次重复,每个重复4头进行为期28天的饲养实验。4个处理组分别为:基础日粮 0.5%果寡糖(FOS组)、基础日粮 0.3%甘露寡糖(MOS组)、基础日粮 0.45%果寡糖 0.25%甘露寡糖(F M组)、基础日粮 75 mg/kg金霉素 40 mg/kg洛克沙生(ABT组)。实验结果表明:(1)寡糖组免疫器官指数较ABT组大(P>0.05)。(2)在断奶后第7天,FOS组(0.32±0.49)、MOS组(0.32±0.04)和F M组(0.32±0.07)均较ABT组(0.20±0.01)显著提高了血清免疫球蛋白A(IgA)的浓度(P<0.05),同时FOS组(3.50±0.49)较ABT组(2.68±0.47)显著提高了IgG浓度(P<0.05);F M组显著提高了断奶后第21天IgG浓度(P<0.05);MOS组显著提高了断奶后第21天IgM浓度(P<0.05)。(3)与ABT组相比,FOS显著提高了断奶后第7天胸腺(21.33±6.03对37.33±4.04)、脾脏(19.67±2.08对30.33±8.73)、淋巴结(24.67±4.16对37.67±2.52)CD4 T淋巴细胞百分数(P<0.05);FOS组(21.33±4.04对32.00±5.29)、MOS组(21.33±4.04对37.33±3.21)、F M组(21.33±4.04对32.00±3.60)显著提高了断奶后第28天胸腺CD4 T淋巴细胞百分数(P<0.05);MOS组(20.67±3.51对29.67±5.51)显著提高了断奶后第7天胸腺CD8 T淋巴细胞百分数;F M组(17.00±1.00对22.67±3.06)显著提高了断奶后第7天脾脏CD8 T淋巴细胞百分数(P<0.05);各寡糖组对断奶后第28天CD4 、CD8 T淋巴细胞百分数影响不显著(P>0.05);ABT组(34.33±5.03)在断奶后第7天较F M组(20.67±6.43)显著提高了淋巴结CD8 T淋巴细胞百分数(P<0.05);在断奶后第7天,FOS组(1.42±0.32)、MOS组(1.52±0.46)、F M组(1.51±0.30)淋巴结CD4 与CD8 比值显著(P<0.05)大于ABT组(0.71±0.03);在断奶后第28天,FOS组(1.36±0.21)和MOS组(1.34±0.16)脾脏CD4 与CD8 比值显著(P=0.01)大于ABT组(0.94±0.29),除此之外,各寡糖组CD4 与CD8 比值较寡糖组大(P>0.05),寡糖组间差异不显著。     

Mannan biotechnology: from biofuels to health

Mannans of different structure and composition are renewable bioresources that can be widely found as components of lignocellulosic biomass in softwood and agricultural wastes, as non-starch reserve polysaccharides in endosperms and vacuoles of a wide variety of plants, as well as a major component of yeast cell walls. Enzymatic hydrolysis of mannans using mannanases is essential in the pre-treatment step during the production of second-generation biofuels and for the production of potentially health-promoting manno-oligosaccharides (MOS). In addition, mannan-degrading enzymes can be employed in various biotechnological applications, such as cleansing and food industries. In this review, fundamental knowledge of mannan structures, sources and functions will be summarized. An update on various aspects of mannan-degrading enzymes as well as the current status of their production, and a critical analysis of the potential application of MOS in food and feed industries will be given. Finally, emerging areas of research on mannan biotechnology will be highlighted.     

Fungal Zymosan and Mannan Activate the Cryopyrin Inflammasome

Some fungal species are opportunistic pathogens that can cause infection in people with compromised immune systems. Activation of caspase-1 and the subsequent secretion of mature interleukin (IL)-1β is a major signaling pathway of the innate immune system, but how yeasts induce caspase-1 activation is unknown. We show here that stimulation of macrophages and dendritic cells with heat-killed Saccharomyces cerevisiae or the purified cell wall components zymosan and mannan induced caspase-1 activation and IL-1β secretion when combined with ATP. Macrophages deficient for the inflammasome adaptor ASC were defective in caspase-1 activation and IL-1β secretion, suggesting involvement of an ASC-dependent inflammasome. Indeed, caspase-1 activation was abrogated in macrophages lacking the NOD-like (NLR) protein Cryopyrin/Nalp3 and in wild type macrophages pretreated with the pannexin-1 inhibitor probenecid. IL-1β secretion further required the Toll-like receptor (TLR) adaptors MyD88 and TRIF, and partially relied on TLR2. We previously showed that bacterial molecules such as lipopolysaccharide (LPS) and peptidoglycan induce activation of caspase-7 through the Cryopyrin inflammasome. Similarly, Cryopyrin and ASC were required for activation of caspase-7 in macrophages stimulated with zymosan or mannan and ATP. These results demonstrate that the conserved fungal components zymosan and mannan require ASC and Cryopyrin for caspase-1 activation and IL-1β secretion and suggest an important role for the Cryopyrin inflammasome during fungal infections.Pathogen recognition by the innate immune system relies on a limited number of fixed germline-encoded receptors, which have evolved to identify so-called pathogen-associated molecular patterns (PAMPs),² conserved microbial structures not shared by the host and essential for their survival (1). Examples of PAMPs are LPS from Gram-negative bacteria, peptidoglycan (PGN) from Gram-positive bacteria, and zymosan and mannan from fungi. Several structurally and functionally diverse classes of pattern-recognition receptors (PRRs) exist that induce various host defense pathways, including the Toll-like receptors (TLRs) located in the plasma membrane and intracellular organelles and the more recently identified intracellular family of NOD-like receptors (NLRs) (2).Previous studies have shown that gain-of-function mutations within the NLR protein Cryopyrin/NALP3 are associated with three autoinflammatory disorders characterized by skin rashes and prolonged episodes of fever in the absence of any apparent infection (3, 4). These hereditary periodic fever syndromes are Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FACS), and neonatal-onset multisystem inflammatory disease (NOMID), and they are collectively referred to as the Cryopyrin/NALP3-associated periodic syndromes (CAPS). Subsequent studies revealed that the Cryopyrin/Nalp3 plays a crucial role in the assembly of a large (700 kDa) cytosolic protein complex termed the “inflammasome” (5–7). The bipartite adaptor protein ASC bridges the interaction between Cryopyrin/Nalp3 and caspase-1 in the inflammasome; thus allowing the recruitment and autoproteolytic activation of the cysteine protease (2). Activated caspase-1 subsequently mediates the maturation and secretion of the pro-inflammatory cytokines interleukin (IL)-1β and IL-18 (8–10). Interestingly, the Cryopyrin/Nalp3 inflammasome mediates caspase-1 activation in response to a variety of bacterial PAMPs such as LPS and PGN when combined with a second stimulus such as the P2X₇ receptor ligand ATP (11–14). Cryopyrin/Nalp3 also mediates caspase-1 activation and IL-1β secretion in macrophages stimulated with viral RNA and ATP (15) or exposed to crystalline substances including uric acid, silica and asbestos (16–18). In contrast, the related NLR protein Ipaf is required for caspase-1 activation in macrophages infected with the intracellular pathogens Salmonella, Legionella, and Shigella (19–21).Although the roles of specific inflammasomes in response to bacterial and viral PAMPs have been described, the inflammasome complexes that recognize fungal PAMPs to induce caspase-1 activation and IL-1β secretion are unknown. Here we show that heat-killed Saccharomyces cerevisiae and the purified cell wall components zymosan and mannan induced caspase-1 activation and IL-1β secretion from macrophages and dendritic cells upon co-stimulation with ATP. Macrophages deficient for the inflammasome adaptor ASC or the NLR protein Cryopyrin/Nalp3 were defective in zymosan- and mannan-induced caspase-1 activation and IL-1β secretion, whereas TNF-α secretion remained unaffected. Although macrophages lacking the TLR adaptors MyD88 or TRIF still activated caspase-1, zymosan- and mannan-induced secretion of IL-1β was significantly hampered. These results demonstrate that the conserved fungal cell wall components zymosan and mannan require ASC and Cryopyrin for caspase-1 activation and IL-1β secretion and suggest an important role for the Cryopyrin inflammasome during fungal infections.     

Mannan synthetase activity in three strains of Candida albicans

Abstract Mannan synthetase activity has been investigated in Candida albicans , strain 4918, as well as in two relatively avirulent, cerulenin-resistant mutant derivative strains, 4918-2 and 4918-10. In addition, investigations pertaining to the effects of the agents, cerulenin and sodium butyrate, on the level of mannan synthetase activity during the yeast to hyphal transition of these strains have been performed. The results show that mannan synthetase activity in yeast cells of both mutant strains is consistently higher than that observed in the parental strain. Similarly, the profile of enzyme activity exhibited by the mutant strains as morphogenesis proceeds differs from that of the wild-type. Sodium butyrate has no significant effect on enzyme activity in these strains, but the presence of cerulenin results in alterations in mannan synthethase activity during morphogenesis of strain 4918.     

Determination of Acidic Component of Konjac Mannan

mRNA was isolated from mammary glands of lactating cow by affinity chromatography on poly(U)-Sepharose. The mRNA was heterogeneous on 3% agarose gel electrophoresis in the presence of 6m urea. The molecular weight of the main peak was estimated to be 3.3 x 10⁵. The mRNA was translated in a cell-free protein synthesizing system derived from wheat germ extract, and the translation products were analysed by the indirect immunoprecipitation method using specific antisera for casein components. About 50% of the total protein directed by this mRNA was casein. The relative amounts of α_s1-, β-,and k-casein in the translation products were nearly the same as those in bovine milk. The immunoprecipitates were analysed on sodium dodecyl sulfatepolyacrylamide gradient gel (15~20%) electrophoresis, and their mobilities were compared with those of dephosphorylated and non-glycosylated casein as standard, α_s1- and k Casein synthesized in vitro migrated more slowly than standard caseins, while synthesized β-casein migrated slightly faster than the standard β-casein.     

Peptization of the Gel of Konjac Mannan

The addition of 1,8-pyrenequinone into the assay system containing rat liver homogenates (S-9) caused an approximately 10-fold increase in the mutagenicity of 2-acetylaminofluorene (AAF) in the current Salmonella reversion assay system. Since no chemical reaction between 1,8-pyrenequinone and AAF was observed, the in vitro effects of 1,8-pyrenequinone on the metabolisms of AAF with S-9 mix were studied. The enhancement of mutagenicity by 1,8-pyrenequinone was not dependent on the dose of NADPH under the present assay condition. The mutagenicity of AAF was increased approximately 4-fold by the addition of 1,8-pyrenequinone into microsomes, whereas it remained at the spontaneous level in the presence of cytosol. However, by reconstituting microsomes with cytosol, the mutagenicity enhancing activity was recovered to the original level. Since 1,8-pyrenequinone inhibited the AAF hydroxylase activity, chemical analysis of the incubation mixture of AAF was tried. This indicated that a higher amount of unmetabolized AAF remained and higher amounts of 2-aminofluorene and N-hydroxy-2-acetylaminofluorene were accumulated in the presence of 1,8-pyrenequinone compared with those in the absence of 1,8-pyrenequinone. From these results, it seems probable that 1,8-pyrenequinone inhibits C-hydroxylation (the detoxifying pathway) and promotes N-hydroxylation (the activating pathway) as well as deacetylation in the AAF metabolism.     

The Mechanism of Gelation of Konjac Mannan

The sol of konjac mannan (KM) was gelatinized with an alkali such as sodium carbonate. The turbidity and the viscosity of sol, the infrared spectra of KM, and the consumption of alkali by KM in the course of gelation were measured. Then, experiments were undertaken in order to elucidate the major role of alkalies in gelation and the mechanism of gel-formation. It was presumed that the alkalies eliminate a moiety containing C=O group (probably an organic acid) from KM, and then the molecules of KM which lost the moiety crystallize in part through a linkage such as hydrogen bonding, and a network structure is formed.     

17年生邓恩桉两个种源木材密度与干缩性研究

通过对邓恩桉两个种源木材基本密度和在湿、气、全干状态下的径、弦向尺寸的测定,研究了不同树干位置的基本密度和干缩率的变异规律,并运用加工工艺中减小木材皱缩和开裂措施(蒸汽调湿处理方法)探索皱缩开裂缺陷的恢复性能。结果表明:1)邓恩桉种源100(0.5265 g/cm³)的木材基本密度小于种源98(0.5360 g/cm³);2)木材基本密度在树干纵向呈波浪式变化,在横向边材显著大于心材;3)木材干缩率在纵向随高度的增加而减小,在横向边材大于心材,且弦向干缩率均为径向的1~3倍;4)经喷蒸调湿处理后,木材开裂程度减轻,且径向和弦向皱缩恢复率分别为0.4%、3.3%;5)木材基本密度与气干弦向干缩率存在极显著负相关(r=-0.313),并建立了气干干缩率与全干干缩率的回归模型。