Digestibility by sheep of direct cut alfalfa silage made with ozonated cotton stalks

Alfalfa was ensiled either by conventional wilting (WA) or as a mixture with 20% (DM) ozonated cotton stalks (A + OCS), and the digestibility of the silage carbohydrate constituents was examined in sheep. The WA silage had a lower content of glucose, xylose, and total carbohydrates and a slightly higher content of arabinose, galactose and mannose than the A + OCS silage. Stomachs and whole tract carbohydrate digestibility was 52% and 63%, respectively, in both silages. Digestibility of neutral detergent fiber (NDF) glucose and xylose was slightly higher and that of NDF-arabinose, galactose and mannose slightly lower, in the sheep fed A + OCS vs. those fed WA.Digestibility of total NDF monosaccharide components in WA and A + OCS silages were 55% and 57%, respectively, of which 90% were digested in the fore-stomachs. In both silages, NDF-glucose was more digestible than NDF-xylose and NDF-uronic acid, and less digestible than NDF-arabinose and NDF-mannose.Results of this study indicate that a direct-cut and ensiled alfalfa silage made with 20% (DM) OCS is comparable with WA in its potential to supply ruminants with digestible carbohydrates.     

Comparison of carbohydrate substrate preferences in eight species of bifidobacteria

Eight species of bifidobacteria were tested for their abilities to grow on a range of monosaccharides (glucose, arabinose, xylose, galactose and mannose). In contrast to the other sugars, glucose and galactose were utilized by all species and, in general, specific growth rates were highest on these sugars. Different substrate preferences were observed between species when the bacteria were grown in the presence of all five monosaccharides. For example, glucose and xylose were coutilized by Bifidobacterium longum, whereas glucose repressed uptake of all other sugars in B. bifidum and B. catenulatum. Galactose was the preferred substrate with B. pseudolongum. In B. angulatum, glucose and galactose were utilized simultaneously. B. breve did not grow on arabinose when this sugar provided the sole source of energy. However, glucose and arabinose were preferentially taken up during growth on sugar mixtures.     

The transport and mediation mechanisms of the common sugars in Escherichia coli

Escherichia coli can uptake and utilize many common natural sugars to form biomass or valuable target bio-products. Carbon catabolite repression (CCR) will occur and hamper the efficient production of bio-products if E. coli strains are cultivated in a mixture of sugars containing some preferred sugar, such as glucose. Understanding the transport and metabolism mechanisms of the common and inexpensive sugars in E. coli is important for further improving the efficiency of sugar bioconversion and for reducing industrial fermentation costs using the methods of metabolic engineering, synthetic biology and systems biology. In this review, the transport and mediation mechanisms of glucose, fructose, sucrose, xylose and arabinose are discussed and summarized, and the hierarchical utilization principles of these sugars are elucidated.     

Fermentation of corn fibre sugars by an engineered xylose utilizing Saccharomyces yeast strain

The ability of a recombinant Saccharomyces yeast strain to ferment the sugars glucose, xylose, arabinose and galactose which are the predominant monosaccharides found in corn fibre hydrolysates has been examined. Saccharomyces strain 1400 (pLNH32) was genetically engineered to ferment xylose by expressing genes encoding a xylose reductase, a xylitol dehydrogenase and a xylulose kinase. The recombinant efficiently fermented xylose alone or in the presence of glucose. Xylose-grown cultures had very little difference in xylitol accumulation, with only 4 to 5g/l accumulating, in aerobic, micro-aerated and anaerobic conditions. Highest production of ethanol with all sugars was achieved under anaerobic conditions. From a mixture of glucose (80g/l) and xylose (40g/l), this strain produced 52g/l ethanol, equivalent to 85% of theoretical yield, in less than 24h. Using a mixture of glucose (31g/l), xylose (15.2g/l), arabinose (10.5g/l) and galactose (2g/l), all of the sugars except arabinose were consumed in 24h with an accumulation of 22g ethanol/l, a 90% yield (excluding the arabinose in the calculation since it is not fermented). Approximately 98% theoretical yield, or 21g ethanol/l, was achieved using an enzymatic hydrolysate of ammonia fibre exploded corn fibre containing an estimated 47.0g mixed sugars/l. In all mixed sugar fermentations, less than 25% arabinose was consumed and converted into arabitol.     

Effect of hydrolysing and oxidizing agents on the composition and degradation of wheat straw monosaccharides

Summary Wheat straw (WS) was treated with 5% sodium hydroxide, ozone and 5% sulfur dioxide at 70°C for 72 h, and the effect of treatments on monosaccharide composition and in vitro degradability by rumen microorganisms was studied. The major sugars, glucose and xylose, comprising about 90% of the total monosaccharides in the untreated WS were mainly confined to the cell walls. SO₂ exerted the greatest solubilizing effect, followed by ozone and NaOH; the respective values for the solubilized cell wall polysaccharides were: 26, 12 and 4.4%. One third of the total phenolics was oxidized by ozone, whereas, SO₂ exerted mostly a solubilizing effect on this fraction, converting 75% of it into soluble phenolics. In the NaOH treated WS 41% of the total phenolics were soluble, as compared to 22% in the untreated. The in vitro digestibility of monosaccharides in the untreated WS were initially high: 50% and 58% for xylose and glucose, respectively and 63% to 80% for the minor sugars. The SO₂ treatment resulted in an overall increase in digestibility of monosaccharides with values lying in the range of 90%. Sodium hydroxide was more efficient than ozone in enhancing the degradability of xylan and total sugars. The digestibility of cell wall sugars was increased from 52.4% to 84.4%, 63.4% and 72.3% by SO₂, O₃ and NaOH treatments respectively. Based on the present findings, it appears that wheat straw cell wall components are more sensitive to hydrolytic than to oxidative processes aimed at increasing its degradability by rumen microorganisms. SO₂ exerted on WS a multi-effect which was particularly suitable for increasing the digestibility of monosaccharides.     

Improved method of analysis of biomass sugars using high-performance liquid chromatography

The precise quantitative analysis of biomass derived sugars is a very important step in the conversion of biomass feedstocks to fuels and chemicals. However, the most accurate method of biomass sugar analysis is based on the gas chromatography analysis of derivatized sugars either as alditol acetates or trimethylsilanes. The derivatization method is time-consuming but the alternative HPLC method cannot resolve most sugars found in biomass hydrolysates. We have demonstrated for the first time that by careful manipulation of the HPLC mobile phase, biomass monomeric sugars (arabinose, xylose, fructose, glucose, mannose, and galactose) can be analyzed quantitatively and there is excellent baseline resolution of all the sugars. This was demonstrated for both standard sugars and corn stover hydrolysates. Our method can also be used to analyze dimmeric sugars (cellobiose and sucrose).     

Synthesis and application of Fmoc-hydrazine for the quantitative determination of saccharides by reversed-phase high-performance liquid chromatography in the low and subpicomole range

A new derivatization reagent, Fmoc-hydrazine, has been synthesized from the reaction of Fmoc-chloroformate with hydrazine as a precolumn fluorometric labeling reagent for reducing sugars such as glucose, galactose, mannose, fructose, fucose, ribose, xylose, arabinose, lactose, and maltose. The optimization of derivatization conditions was examined in detail. Using a reversed-phase high-performance C-8 column and a mobile phase consisting of acetonitrile-aqueous acetic acid, seven sugar derivatives were separated under either isocratic or gradient conditions within 20 min. The Fmoc-hydrazine and sugar Fmoc-hydrazone derivatives exhibit excellent stability. The extent of the hydrazone formation was 77 and 82% for mannose and fucose as assessed by Dionex high-performance anion-exchange chromatography with pulsed amperometric detection. Linear calibration graphs were established in the range from 0.5 to 2 pmol and 12 to 110 pmol for individual sugar derivatives. The determination limits were 0.05-0.09 pmol for mannose, galactose, and ribose; 0.1 pmol for maltose, xylose, and glucose; 0.2 pmol for fucose and lactose; 0.3 pmol for arabinose; and 0.4 pmol for fructose. The component monosaccharides of ultramicroquantities of two glycoproteins (e.g., from 7 ng fetuin and ovalbumin) were determined in the subpicomole range.     

Studies on the Carbohydrates of Horse Radish,Cochlearis Armoracia L

In the root of horse radish, sucrose, fructose, glucose and two unknown fructose-oligosac-charides were found in the form of free sugars, in the decreasing order just given, and large amounts of starch, hot 50% methanol-soluble polysaccharide, water-soluble polysaccharide, pectin, hemicellulose, cellulose and a small amount of lignin were found as polysaccharides. As the constructive sugar of these polysaccharides, the following, i.e., glucose, fructose, galactose, arabinose, galacturonic acid and xylose were detected.     

Antennal sugar sensitivity in the click beetle Agriotes obscurus

The occurrence of salt‐, sugar‐sensitive neurones and a mechanoreceptor neurone in the antennal hair‐like gustatory sensilla of the click beetle Agriotes obscurus L. (Coleoptera, Elateridae) is demonstrated using the electrophysiological sensillum tip‐recording technique. The stimulating effect of 13 water soluble sugars at 100 mm is tested on the neurones of these sensilla. Sucrose and fructose are the two most stimulating sugars for the sugar‐sensitive neurone, evoking almost 30 spikes s^?1 at 100 mm . The stimulating effect of arabinose, glucose, mannose, maltose and raffinose is three‐ to five‐fold lower, in the range 5.9–9.6 spikes s^?1. The remaining six sugars, xylose, galactose, rhamnose, cellobiose, trehalose and lactose, have very low (<1 spikes s^?1) or no ability to stimulate the sugar‐sensitive neurone. Concentration/response curves of the sugar‐sensitive neurone to sucrose, fructose and glucose at 0.01–100 mm overlap to a large extent in hibernating, cold reactivated and reproductively‐active beetles. A remarkable 9–50% decrease in the number of spikes evoked by 100 mm fructose and 10–100 mm sucrose occurs, however, in reproductively‐active beetles in June compared with beetles at the beginning of hibernation in October. These findings show that A. obscurus is capable of sensing a wide range sugars via their antennal gustatory sensilla.     

Digestibility of carbohydrates in growing pigs: a comparison between the t-cannula and the steered ileo-caecal valve cannula

We compared the determination of ileal and total tract digestibility of carbohydrates in five experimental diets using a double 5?×?5 Latin square design involving a total of 10 cannulated pigs; half of the pigs were equipped with a simple T-cannula and the other half with steered ileo-caecal valve (SICV)-cannula. The diets consisted of nitrogen-free mixture and soya bean meal, sunflower meal, peas or rape seed cake diluted to about 180?g/kg DM protein with the nitrogen-free mixture. There was no significant difference in the digestibility values using the two types of cannulas with regard to organic matter, sugars (sum of glucose, fructose and sucrose), α-galactosides (sum of raffinose, stachyose and verbascose), starch, cellulose, total non-cellulosic polysaccharides (NCP) and insoluble NCP constituents. The digestibility values for the NCP residues arabinose and galactose, however, were estimated higher but with a lower variability with the SICV-cannula compared with the T-cannula. The type of cannula did no influence the estimation of the total tract digestibility for any of the major dietary constituents, but the total tract digestibility was slightly more variable when the pigs were equipped with the SICV-cannula compared with the T-cannula. There was no difference in the ileal digestibility of sugars, α-galactosides, cellulose and the NCP arabinose and uronic acids residues among the experimental diets, while the ileal digestibility of starch and the remaining NCP sugar residues varied between diets. The total tract digestibility was complete for sugars, α-galactosides and starch, whereas the digestibility of the cell wall constituents varied in accordance with the polymeric composition of the cell walls. It was concluded that ileal digesta samples from SICV-cannula are more homogenous than those from the T-cannula. In cases where the precision of each determination is crucial, the SICV-cannula should be the option.     

Digestibility of carbohydrates in growing pigs: a comparison between the T-cannula and the steered ileo-caecal valve cannula

We compared the determination of ileal and total tract digestibility of carbohydrates in five experimental diets using a double 5 x 5 Latin square design involving a total of 10 cannulated pigs; half of the pigs were equipped with a simple T-cannula and the other half with steered ileo-caecal valve (SICV)-cannula. The diets consisted of nitrogen-free mixture and soya bean meal, sunflower meal, peas or rape seed cake diluted to about 180 g/kg DM protein with the nitrogen-free mixture. There was no significant difference in the digestibility values using the two types of cannulas with regard to organic matter, sugars (sum of glucose, fructose and sucrose), alpha-galactosides (sum of raffinose, stachyose and verbascose), starch, cellulose, total non-cellulosic polysaccharides (NCP) and insoluble NCP constituents. The digestibility values for the NCP residues arabinose and galactose, however, were estimated higher but with a lower variability with the SICV-cannula compared with the T-cannula. The type of cannula did no influence the estimation of the total tract digestibility for any of the major dietary constituents, but the total tract digestibility was slightly more variable when the pigs were equipped with the SICV-cannula compared with the T-cannula. There was no difference in the ileal digestibility of sugars, alpha-galactosides, cellulose and the NCP arabinose and uronic acids residues among the experimental diets, while the ileal digestibility of starch and the remaining NCP sugar residues varied between diets. The total tract digestibility was complete for sugars, alpha-galactosides and starch, whereas the digestibility of the cell wall constituents varied in accordance with the polymeric composition of the cell walls. It was concluded that ileal digesta samples from SICV-cannula are more homogenous than those from the T-cannula. In cases where the precision of each determination is crucial, the SICV-cannula should be the option.     

Preferences and apparent digestibilities of sugars by fruit damaging birds in Japan

Gray starlings Sturnus cineraceus, azure-winged magpies Cyanopica cyana and brown-eared bulbuls Hypsipetes amaurotis are among the main bird pests in commercial fruit orchards in central Japan. Recently Brugger & Nelms (1991) suggested that developing high-sucrose fruit cultivars could reduce crop damage, because some pest birds lack the enzyme sucrase and can develop an aversion to sucrose. Preferences for, and digestibilities of, the monosaccharides glucose and fructose and the disaccharide sucrose by these pests species were therefore studied to assess whether this idea would be applicable in central Japan. Gray starlings and brown-eared bulbuls were able to detect glucose, fructose, a mixture of glucose and fructose, and sucrose at a concentration of 12% w/v. Azure-winged magpies also detected glucose and fructose, but failed to detect sucrose at the same concentration. In pairwise preference trials gray starlings and azure-winged magpies selected the monosaccharides over sucrose, but brown-eared bulbuls did not. To estimate the digestibility of the sugars the apparent assimilated mass coefficient, AMC*, was calculated for each species eating each sugar by measuring intake and faecal output. Monosaccharides had mean AMC*s of 0.77, 0.96 and 0.92 when consumed by gray starlings, azure-winged magpies and brown-earned bulbuls respectively. AMC* values for sucrose were 0.82 and 0.49 for brown-eared bulbuls and azure-winged magpies respectively, but gray starlings were shown to be unable to digest sucrose. As AMC* values varied from 0.75 to 0.97, consumption rates of sugars increased as digestibility decreased. Although increasing sucrose contents of commercial fruits may deter sucrase-deficient birds such as gray starlings from depredating fruits, it may also lead to increased crop damage by species such as the azure-winged magpie and brown-eared bulbul which may have to consume more of the less digestible fruit in order to meet their energy requirements.     

Role of catabolite regulatory mechanisms in control of carbohydrate utilization by the rumen anaerobic fungus Neocallimastix frontalis.

Neocallimastix frontalis PN-1 utilized the soluble sugars D-glucose, D-cellobiose, D-fructose, maltose, sucrose, and D-xylose for growth. L-Arabinose, D-galactose, D-mannose, and D-xylitol did not support growth of the fungus. Paired substrate test systems were used to determine whether any two sugars were utilized simultaneously or sequentially. Of the paired monosaccharides tested, glucose was found to be preferentially utilized compared with fructose and xylose. The disaccharides cellobiose and sucrose were preferentially utilized compared with fructose and glucose, respectively, an cellobiose was also the preferred substrate compared with xylose. Xylose was the preferred substrate compared with maltose. In further incubations, the fungus was grown on the substrate utilized last in the two-substrate tests. After moderate growth was attained, the preferred substrate was added to the culture medium. Inhibition of nonpreferred substrate utilization by the addition of the preferred substrate was taken as evidence of catabolite regulation. For the various combinations of substrates tested, fructose and xylose utilization was found to be inhibited in the presence of glucose, indicating that catabolite regulation was involved. No clear-cut inhibition was observed with any of the other substrate combinations tested. The significance of these findings in relation to rumen microbial interactions and competitions is discussed.     

Carbohydrate utilization patterns and substrate preferences in Bacteroides thetaiotaomicron

Specific growth rates of Bacteroides thetaiotaomicron NCTC 10582 with either glucose, arabinose, mannose, galactose or xylose as sole carbon sources were 0.42/h, 0.10/h, 0.38/h, 0.38/h and 0.16/h respectively, suggesting that hexose metabolism was energetically more efficient than pentose fermentation in this bacterium. Batch culture experiments to determine whether carbohydrate utilization was controlled by substrate-induced regulatory mechanisms demonstrated that mannose inhibited uptake of glucose, galactose and arabinose, but had less effect on xylose. Arabinose and xylose were preferentially utilized at high dilution rates (D > 0.26/h) in carbon-limited continuous cultures grown on mixtures of arabinose, xylose, galactose and glucose. When mannose was also present, xylose was co-assimilated at all dilution rates. Under nitrogen-limited conditions, however, mannose repressed uptake of all sugars, showing that its effect on xylose utilization was strongly concentration dependent. Studies with individual D-ZU-14C]-labelled substrates showed that transport systems for glucose, galactose, xylose and mannose were inducible. Measurements to determine incorporation of these sugars into trichloroacetic acid-precipitable material indicated that glucose and mannose were the principal precursor monosaccharides. Xylose was only incorporated into intracellular macromolecules when it served as growth substrate. Phosphoenolpyruvate:phosphotransferase systems were not detected in preliminary experiments to elucidate the mechanisms of sugar uptake, and studies with inhibitors of carbohydrate transport showed no consistent pattern of inhibition with glucose, galactose, xylose and mannose. These results indicate the existence of a variety of different systems involved in sugar transport in B. thetaiotaomicron.     

Role of catabolite regulatory mechanisms in control of carbohydrate utilization by the rumen anaerobic fungus Neocallimastix frontalis

Neocallimastix frontalis PN-1 utilized the soluble sugars D-glucose, D-cellobiose, D-fructose, maltose, sucrose, and D-xylose for growth. L-Arabinose, D-galactose, D-mannose, and D-xylitol did not support growth of the fungus. Paired substrate test systems were used to determine whether any two sugars were utilized simultaneously or sequentially. Of the paired monosaccharides tested, glucose was found to be preferentially utilized compared with fructose and xylose. The disaccharides cellobiose and sucrose were preferentially utilized compared with fructose and glucose, respectively, an cellobiose was also the preferred substrate compared with xylose. Xylose was the preferred substrate compared with maltose. In further incubations, the fungus was grown on the substrate utilized last in the two-substrate tests. After moderate growth was attained, the preferred substrate was added to the culture medium. Inhibition of nonpreferred substrate utilization by the addition of the preferred substrate was taken as evidence of catabolite regulation. For the various combinations of substrates tested, fructose and xylose utilization was found to be inhibited in the presence of glucose, indicating that catabolite regulation was involved. No clear-cut inhibition was observed with any of the other substrate combinations tested. The significance of these findings in relation to rumen microbial interactions and competitions is discussed.     

马克斯克鲁维酵母GX-UN120糖转运蛋白基因的克隆及表征

【背景】马克斯克鲁维酵母(Kluyveromyces marxianus)具有完整的木糖代谢途径,可以高效利用木质纤维素中的木糖,因此对其糖转运蛋白基因的研究或可有效解决酵母木糖转运的相关问题。【目的】根据马克斯克鲁维酵母DMKU3-1042中KLMA＿70145和KLMA＿80101基因位点的功能预测,获得马克斯克鲁维酵母GX-UN120相应的糖转运蛋白基因序列并探究其功能。【方法】将转运蛋白基因分别克隆表达至酿酒酵母EBY.VW4000中考察重组菌株生长特性,以此间接评价对应转运蛋白的转运能力。【结果】Km＿SUT2基因编码的糖转运蛋白可有效提高宿主细胞转运木糖、阿拉伯糖、山梨糖、核糖、乳糖和葡萄糖的能力,但却不能转运甘露糖、果糖、蔗糖和半乳糖。类似地,Km＿SUT3基因编码的糖转运蛋白可提高细胞转运木糖、阿拉伯糖、山梨糖、半乳糖、核糖、乳糖和葡萄糖的能力,但却不能转运甘露糖和果糖。然而在葡萄糖存在的条件下,重组菌株对各种碳源的利用均受抑制,但Km＿SUT3转运木糖和核糖过程中受葡萄糖的抑制作用较小。【结论】马克斯克鲁维酵母GX-UN120中转运蛋白Km＿SUT2和Km＿SUT3可...     

中性糖在土壤中的来源与分布特征

糖类(即碳水化合物)是土壤有机质的重要组成部分, 经生物化学降解形成不同结构的单糖。土壤中的中性单糖也叫中性糖, 主要包括木糖、核糖、阿拉伯糖、葡萄糖、半乳糖、甘露糖、岩藻糖和鼠李糖。其中, 植物来源的糖主要为五碳糖, 如木糖和阿拉伯糖; 微生物来源的糖主要包括半乳糖、甘露糖、岩藻糖、鼠李糖等六碳糖。研究中常利用六碳糖和五碳糖的比例指示微生物和植物对土壤有机碳的相对贡献。中性糖是微生物重要的碳源和能量来源, 在团聚体的形成过程中扮演着重要角色。该文整合了近30年土壤中性糖的研究进展, 对比了提取中性糖的常用方法, 分析了不同土地利用类型和不同土壤组分中中性糖的含量、来源和周转特征, 综述了影响中性糖含量和分布的主要环境因素。结果表明, 中性糖在耕地土壤中的绝对含量和相对含量均显著低于针叶林、阔叶林、草地和灌丛4种土地利用类型。(半乳糖+甘露糖)/(阿拉伯糖+木糖)(GM/AX)在不同土地利用间差异不显著, 而(鼠李糖+岩藻糖)/(阿拉伯糖+木糖)(RF/AX)则表明草地土壤中的微生物来源的中性糖含量高于针叶林和耕地。不同密度的土壤组分中, 轻质组分中中性糖的含量比重质组分高, 重质组分中微生物来源的中性糖较多; 就不同粒径(或团聚体)而言, 黏粒(或微团聚体)中微生物来源的中性糖含量更丰富。有关影响土壤中性糖含量和分布的因素的研究, 目前主要集中在人为活动(如耕种和放牧等), 而有关温度、降水等自然环境因素影响的研究较少。     

Golgi Apparatus Mediated Polysaccharide Secretion by Outer Root Cap Cells of Zea mays. III. Control by Exogenous Sugars

Sugars supplied to germinating seedlings of maize (Zea mays L.) regulate the secretion of polysaccharides by the outer cells of the root cap. The polysaccharide secreted by these cells adheres to the root tip as a droplet and the size of the droplet was used to quantitate polysaccharide secretion. The polysaccharide contains glucose, galacrose, and galacturonic acid residues with smaller quantities of mannose, arabinose, xylose, fucose and rhamnose. These sugars supplied to maize seedlings had marked effects on the rate of polysaccharide secretion by root tips. The effects on secretion were independent of the growth rates of the roots. Glucose, fucose and xylose increased droplet size 1.5–2 fold (as did sucrose, maltose, lacrose, fructose and ribose) whereas galactose, arabinose and galacturonic acid were inhibitory. Mannose increased dropler size 5–7 fold. The marked effect of mannose on polysaccharide secretion was due to an increased rate of secretion combined with a longer phase of extrusion of polysaccharide into the forming droplet. The effect of mannose was partially reversed by inorganic phosphate and other sugars (except for fucose which had no effect or promoted secretion in the presence of mannose). In contrast to sucrose, mannose stimulated secretion in a maize variety having a high sugar endosperm (high endogenous sugar). The results suggest that regulation of secretion by mannose is due to an alteration of normal sugar metabolism; whereas stimulation of secretion by sucrose and other sugars may be due to an increased availability of sugars for metabolism.