Higher Saccharides Research Articles

Combined Microbial Fermentation Converts Bioactive Compounds in Nitraria Tangutorum Bobrov Fruit and Displays its Antidiabetic Potential

Nitraria tangutorum Bobrov is a berry shrub with white flowers and red fruits, which grows in the deserts in the Tibetan Plateau, Mongolia, and western China. Its fruit N. tangutorum Fruit (NTF) contains various bioactive compounds, with anti-fatigue, anti-inflammatory, and neuroprotective functions. However, the high saccharide content of NTF makes it unsuitable for diabetic patients. In this study, we fermented NTF to obtain N. tangutorum Fermented Juice (NTJ), and N. tangutorum Fermented Residue (NTR), which are suitable for diabetics to consume. We characterized the bioactive compounds in NTF, NTR, and NTJ, and found that fermentation increased the diversity of bioactive compounds, and greatly reduced sucrose, glucose, and fructose content while generating trehalose, which has the potential to manage blood glucose levels. Further, NTJ displays anti-diabetic potential due to various compounds anti-diabetic properties. This study provides a basis for further clinical research on NTJ’s anti-diabetic function in humans.

Enzymatic liquification pattern of superior sweet potato starch of CIP-type

The liquification pattern of each type of starch at the best temperature and enzyme concentration varies, and the physical and chemical properties influence it. The whiter the starch, the smaller and more uniform the size of the granule and the higher amylose level and more amorphous regions, thus it results in a better liquification pattern and feasibility of starch for industrial raw materials. This research aimed to study the best changes in saccharide pattern during the liquification process of superior Centro International de La Papa (CIP)-type sweet potato starch at a temperature of 85oC and the concentration of alpha-amylase enzyme (0.07%) obtained from preliminary research. Hence, the advantages of the resulting hydrolyzate products were compared. The sweet potato starches tested were CIP-1, CIP-2, and CIP-5, tested using the Completely Randomized Design method with two repetitions. The variables observed were DE, dextrin, changes in amylase and amylopectin levels, high saccharide levels, maltotriose, maltose, glucose, and viscosity. The results showed that liquification of CIP-1 sweet potato starch for 225 mins produced better hydrolysate (30.52% DE, 16.28% maltose, 18.70% maltotriose, 1.06% glucose). Meanwhile, 20 mins of liquification resulted in higher dextrin (84.86%). However, the viscosity of CIP-1 was slightly thicker (22 cPs) than CIP-2 (15 cPs) and CIP -5 (15 cPs). The results showed that three types of starch were qualified for raw materials in the advanced hydrolysis industry (DE value was in the range of 20 - 45%), such as saccharification, with the order that CIP-1 was better than CIP-2 and CP-2 was better than CIP-5.

Effects of Hydrothermal Processing on Miscanthus × giganteus Polysaccharides: A Kinetic Assessment.

Miscanthus × giganteus samples were characterized for composition and treated with hot compressed water (hydrothermal or autohydrolysis treatments) at temperatures in the range of 190-240 °C. The liquid phases from treatments were analyzed to assess the breakdown of susceptible polysaccharides into a scope of soluble intermediates and reaction products. The experimental concentration profiles determined for the target compounds (monosaccharides, higher saccharides, acetic acid and sugar-decomposition products) were interpreted using a pseudohomogeneous kinetic mechanism involving 27 reactions, which were governed by kinetic coefficients showing an Arrhenius-type temperature dependence. The corresponding activation energies were calculated and compared with data from the literature. The kinetic equations allowed a quantitative assessment of the experimental results, providing key information for process simulation and evaluation.

Marine Bacteria Affect Saccharide Enrichment in Sea Spray Aerosol during a Phytoplankton Bloom

Saccharides are a large class of organic matter in sea spray aerosol (SSA) that can impact its climate-relevant properties. In seawater, saccharides are produced, exuded, and consumed by phytoplankton and heterotrophic bacteria (HB). Herein, we add the HB strains Alteromonas sp. (AltSIO), Pseudoalteromonas (ATW7), and Flavobacteria bacterium (BBFL7) to marine microcosms during phytoplankton blooms and directly probe the impact of HB on the composition and size of saccharides in seawater and SSA for the first time. Enrichment factors (EFs) for saccharide SSA relative to seawater were composition- and size-dependent, ranging from 100 to 930 000 in sub-micrometer SSA and from 38 to 3700 in super-micrometer SSA. For saccharides in two similar phytoplankton blooms, lower seawater concentrations coincided with higher EFs, which may explain high saccharide enrichments in remote marine locations. Increased enzyme activity within a day of HB addition corresponded to increases in saccharide EFs in sub-micrometer SSA, by a factor of 30 for the energy-storage saccharide glucose and by a factor of 2 for the structural saccharide xylose. Meanwhile, arabinose, released in response to microorganism stress, increased by a factor of 20 in super-micrometer SSA after a lag of 2 days. Observations of HB in SSA indicate that modifications to the saccharide pool in terms of composition and size occur in both seawater and SSA, which subsequently impact their enrichment in SSA. A better understanding the chemical, biological, and physical factors that contribute to the large enrichment of saccharides in SSA will advance our ability to predict SSA composition and its impact on Earth’s climate.

Improving saccharide concentration by mixing octyl acetate during semi-flow, hot-compressed water treatment of woody biomass

Biomass-based ethanol or acetic acid can be produced by fermentation of saccharides following wood hydrolysis. Both high saccharide yield and concentration are essential for achieving an efficient hydrolysis process. However, there can be a trade-off between yield and concentration during semi-flow, hot-compressed water treatment, a candidate for wood hydrolysis. This trade-off is because the fast water flow rate is necessary for high saccharide yields, but the large amount of water dilutes the produced hydrolysate. In this study, we mixed octyl acetate, which is a water-immiscible solvent, during hot-compressed water treatment of beech wood. By adding octyl acetate, a high flow rate was achieved without increasing the water flow rate, and the saccharide concentration was thus improved, while suppressing the decrease in saccharide yield. Furthermore, lignin-derived oligomers, which have inhibitory effects on fermentation, were removed from hydrolysate because they can dissolve in octyl acetate. We performed acetic acid fermentation using the obtained hydrolysate and found that the fermentability was markedly improved compared with the hydrolysate obtained without octyl acetate.

Characterization of Aqueous Products Obtained from Hydrothermal Liquefaction of Rice Straw: Focus on Product Comparison via Microwave-Assisted and Conventional Heating

This paper focuses on the comparison of aqueous products obtained from hydrothermal liquefaction (HTL) of rice straw via microwave (MW)-assisted and conventional heating treatments. A systematic investigation of HTL experiments of rice straw via MW-assisted and conventional heating treatments has been carried out, covering a broad but mild temperature range from 150 to 230 °C at 20 °C intervals. In addition, different reaction times were studied when the HTL reaction temperature was 210 °C. In comparison to conventional HTL, considerable aqueous products could be obtained for MW-assisted HTL, which consumes less time, and in addition, repolymerization behavior could be efficiently decreased and high saccharide yields could be obtained with MW-assisted heating. Besides, the HTL temperature appeared to be the dominant factor, while the increased residence time slightly changed the contents of total organic carbon (TOC) and typical sugars and acids for both conventional and MW-assisted HTL heating.

Lignin cationization with glycidyltrimethylammonium chloride aiming at water purification applications

Hardwood organosolv lignin (OSL) and enzymatic hydrolysis lignin (EHL) from softwood were cationized by glycidyltrimethylammonium chloride (GTAC) as new lignin starting materials for the reaction. The products were in detail characterized by 31P-NMR to elucidate the reactivity of different lignin functionalities in cationization with GTAC. For OSL, a high cationization level (degree of substitution 0.74) was reached leading to a water-soluble product. For EHL, low solubility and lower reactivity were observed, likely due to the high saccharide content. Further, lower reactivity of the guaiacyl type lignin present in softwood, compared to syringyl type lignin in hardwoods, which was shown to react efficiently during the derivatization of OSL, probably played a role. In parallel to cationization, an increase of carboxylic acids (from 0.03 up to 0.55mmol/g) in the lignin was observed as an unexpected side reaction, possibly as a result of alkaline oxygen oxidation. The applicability of the cationized lignins was tested for water purification applications in three model systems which have only sparingly been studied with cationized lignins. GTAC-OSL and GTAC-EHL were found promising for sulfate removal, with sulfate sorption capacities of up to 54mg/g. GTAC-OSL was also found suitable for promoting kaolin settling and to some extent for humic acid coagulation.

Antifungal Activity of Protamine Salmine Hydrochloride and ε-Poly-l-Lysine in Actual Food Systems, Rice- or Wheat-Based Confectioneries

Fungal contamination often occurs in confectioneries with high saccharide content during storage. Protamine salmine hydrochloride (PSH) and ε-poly-l-lysine (ε-PL) showed stable antifungal activities in the medium supplemented with 50% (w/v) sucrose, which pH shows around neutral, as a model of confectioneries. The antifungal activities of PSH and ε-PL have also shown synergy with sodium acetate. To confirm these antifungal activities into actual food systems, dumplings and steamed breads were used as rice- or wheat-based confectionery models. They substantiated practicable antifungal activities against confectionery pollution fungus in these models. The heat stability of both PSH and ε-PL was also indicated, and the impact on the taste or flavor of these confectioneries by addition of PSH and ε-PL were confirmed as less. The results proposed that both PSH and ε-PL are effective antifungal agents in actual foods with high saccharide contents such as confectioneries, at the neutral pH conditions. Practical Applications There are practically few natural antifungal agents used effectively to the actual foods at neutral pH, because many natural antifungal agents are lipophilic, or some of them show the antifungal effects only in the acidic condition even though they are hydrophilic. We screened the natural, hydrophilic antifungal agents effectual at neutral pH by using rice- or wheat-based confectioneries as models, which have actually problems of contamination of fungus. PSH and ε-PL were shown to be effectiveness in those models. They have been known to show strong antimicrobial effects at neutral pH, but they were thought to be necessary to add a large amount into foods to inhibit the growth of fungus. In this study, we demonstrated that the PSH and PL could use as effective antifungal agents with small amount at a high sugar concentration conditions around neutral pH. They were considered to suit for the application to the confectioneries.

Seasonal Variation of Saccharides and Furfural in Atmospheric Aerosols at a Semi-Urban Site

Seasonal variability of daily particle mass and saccharides and furfural concentrations in atmospheric particulate matter in both coarse aerosols (diameter > 2.4 µm) and fine aerosols (diameter < 2.4 µm) was determined for a semi-urban area in the United Kingdom. Saccharides, which include levoglucosan, and furfural are derived from biomass burning and contribute to aerosol composition. This study examined the potential of saccharides and furfural as tracers for biomass combustion. High saccharide concentrations were observed in the autumn, but they did not show a high correlation with potassium expected in biomass smoke. These results may imply that the high saccharide concentrations are derived not only from biomass burning sources, but also from non-combustion sources, such as leaf decay. Significant seasonal variations were observed for saccharides and furfural species in fine atmospheric aerosols. Furfural is likely to be oxidized quickly in comparison with saccharides, so while saccharides such as levoglucosan are known to be fairly stable in the atmosphere, furfural could be transformed. Trajectory and factor analysis suggest that the saccharides may result from long-range transport, while furfural may be more influenced by the local sources possibly because of its shorter lifetime. This result may suggest it could give clues as to the age of biomass smoke.

ChemInform Abstract: Iodine Promoted Simple Synthesis of Benzimidazole Acyclonucleosides.

In nature, 5,6-dimethyl-1-(α-D-ribofuranosyl)benzimidazole exists as a moiety of vitamin B12 (Ref 1). The furanosyl benzimidazole can be considered as Cnucleoside mimic. Due to its structural similarity to purine, benzimidazole is also an important chemical entity in pharmaceuticals 2 . In one approach, aldobenzimimdazoles are prepared by the condensation of aldonic acid or aldonic δ-lactones with o-phenylenediamine 3 . The condensation reactions are often conducted in strong acidic conditions at elevated temperature (e.g.135°C) 4 . The teritol-1-yl benzimidazoles can be further converted to furanosyl benzimidazoles by Lewis acid promoted dehydrative cyclization 5 . In another approach, aldoses are subjected to oxidative condensation with o-phenylenediamine to give aldobenzimidazoles in a direct manner 6 . This approach mitigates the laborious work in the prior preparation of aldonic acids from the corresponding aldoses 7 . For example, Moore and Link 4 have previously applied this approach to synthesize two monosaccharides benzimidazoles, D-gluco and Dgalactobenzimidazoles, in 24% and 50% yields from D-glucose and D-galactose, respectively, by heating with cupric acetate (2 equiv) at 53°C for 14 hr in aqueous acetic acid solution. Some saccharide bisSchiff bases and quinaxolines also form as side products under such reaction conditions 4-6 . However using catalytic amounts of CuSO4 or replacing the AcOH solvent with MeOH does not give any desired aldo-benzimidazoles 8 . Considering the acid-sesitive internal glycosidic bonds in disaccharides and higher saccharides, it is questionable that the corresponding aldo-benimidazoles can be obtained by the above mentioned reactions involving acidic conditions at high reaction temperature. In continuation of our study, the novel transformation of aldehyde and aldoses by use of amines and iodine 9-12 , we describe herein an improved protocol for the synthesis of various aldo-benzimidazoles by direct oxidative condensation of aldoses, including different mono-saccharides with substituted ophenylenediamine in the presence of iodine.

Integrated supercritical fluid extraction and subcritical water hydrolysis for the recovery of bioactive compounds from pressed palm fiber

Pressed palm fiber (PPF), a residue obtained from palm oil industry, is a source of bioactive compounds, such as carotenoids, which are used as food additives. It also has cellulose and hemicellulose that can be used to yield fermentable sugars for the production of second generation ethanol. Supercritical fluid extraction (SFE) of pressed palm fiber provides an oil rich in carotenoids while subcritical water hydrolysis (SubWH) produces hydrolysates with high amounts of fermentable sugars. In this work, the effects of pressure (15–30MPa) and temperature (318 and 328K) on SFE of carotenoids were investigated. The SFE extract with highest carotenoid content was obtained at 318K and 15MPa (2.3% d.b., 0.81mg β-carotene/g extract). After the extraction, the influence of process temperature (423–633K), pressure (15 and 25MPa), solvent:feed ratio (120 and 240), and residence time (1.25–5min) on SubWH of the extraction residue was studied. At the temperature of 523K, the highest total reducing sugar yield (11–23g glucose/100g carbohydrate) and the highest biomass conversion (40–97%) were obtained for any pressure and solvent:feed ratio. The highest selectivity for saccharide formation was found at 423K (20–59mol glucose/mol furfural equivalent). Optimal conditions for high saccharide formation and low sugar degradation product in subcritical hydrolysis were obtained at 523K, 15MPa, solvent:feed ratio of 120, residence time of 2.5min with a total reducing sugar yield of 22.9g glucose/100g carbohydrate and a conversion of 84.9%.

Abstract 583: Ghrelingene expression avoiding the D-ribose caused HepG2 cell death.

Abstract Liver cancer patients could suffer from high saccharide environment due to the malfunction of liver. To mimic this high saccharide environment, HepG2 cells were culture with 30 mM D-riboses. We found that this high concentration of saccharide cause cell death. Cinnamon was report to reduce glucose in model rats. We hypothesized that the administration of cinnamon extracts may benefit liver cancer patients whose metabolism is defected. To test the hypothesis, three different chemical subtypes of cinnamon extracts were added into the culture medium. The adding of all three cinnamon extracts protected HepG2 cells from D-ribose administration and increased around 20% of surviving cells. These protections were even better than the addition of aminoguanidine hemisulfate salt, a chemical used in the treatment of diabetes. The expressions of ghrelin gene and two ghrelin-related genes was monitored by RT-PCR. Ghrelin gene expression was up-regulated in cinnamon treated cells. Also, the ghrelin modify gene, MBOAT4, and ghrelin process enzyme, furin, gene were all detected in HepG2 cells and cinnnamon treated cells. These results demonstrated that if ghrelin expressed in HepG2 cell and maybe liver as well, it can be processed properly. Both ghrelin transcript variants 1 and 3 recombinant plasmid were transfected into HepG2 cells and both transcript variants protected HepG2 cells from the addition of D-ribose without the administration of cinnamon extracts. We conclude that the cinnamon avoids HepG2 cell damage from D-ribose induced oxidative stress by inducing ghrelin gene expression, and through the ghrelin signaling increases cell surviving. Since ghrelin level is abnormally regulated in patients with hepatocellular carcinoma and post-hepatitic liver cirrhosis, the understanding of how ghrelin been regulated in liver may benefit liver disease patients. Citation Format: Jia Huei Shiu, Chin Hao Huang, Tai Lin Lee, Shih Chieh Lee, Shu Ying Liu, Meng Feng Tsai. Ghrelingene expression avoiding the D-ribose caused HepG2 cell death. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 583. doi:10.1158/1538-7445.AM2013-583

Biomineralization in diatoms—phosphorylated saccharides are part of Stephanopyxis turris biosilica

Diatoms—unicellular algae with silicified cell walls—have become model organisms for investigations of biomineralization processes. Numerous studies suggest the importance of biosilica-associated or even embedded biomolecules for the biosilica formation. Such molecules are peptides, polyamines, and even saccharides. However, the role of the latter class of biomolecules is only poorly understood yet. Therefore, we investigated the saccharide composition of the biosilica-associated organic material of the diatom Stephanopyxis turris. This species exhibits a considerably high saccharide content in its siliceous cell walls. Gas chromatography-mass spectrometry analysis revealed that mannose-6-phosphate is strongly associated to the cell walls. This phosphorylated saccharide has not yet been found in diatom biosilica. In vitro studies on the polyallylamine-induced silica precipitation were carried out in the presence of mannose-6-phosphate. Compared to inorganic phosphate, mannose-6-phosphate significantly influenced the precipitation behavior of this model system suggesting a possible contribution of mannose-6-phosphate to the biomineralization process of Stephanopyxis turris.

An optimized approach for enrichment of glycoproteins from cell culture lysates using native multi‐lectin affinity chromatography

Lectins are capable of recognizing specific glycan structures and serve as invaluable tools for the separation of glycosylated proteins from nonglycosylated proteins in biological samples. We report on the optimization of native multi-lectin affinity chromatography, combining three lectins, namely, concanavalin A, jacalin, and wheat germ agglutinin for fractionation of cellular glycoproteins from MCF-7 breast cancer lysate. We evaluated several conditions for optimum recovery of total proteins and glycoproteins such as low pH and saccharide elution buffers, and the inclusion of detergents in binding and elution buffers. Optimum recovery was observed with overnight incubation of cell lysate with lectins at 4°C, and inclusion of detergent in binding and saccharide elution buffers. Total protein and bound recoveries were 80 and 9%, respectively. Importantly, we found that high saccharide strength elution buffers were not necessary to release bound glycoproteins. This study demonstrates that multi-lectin affinity chromatography can be extended to total cell lysate to investigate the cellular glycoproteome.

High dietary fructose intake: Sweet or bitter life?

Epidemiological data show that the consumption of added sugars as ingredients in processed or prepared foods and caloric beverages has dramatically increased. Fructose and fructose-based sweeteners are the most commonly added sugars and high-fructose corn syrup (HFCS-55: 55% fructose, 42% glucose and 3% higher saccharides) accounts for over 40% of all added caloric sweeteners. Concerns regarding the health risk of added sugar follow the demonstration that the consumption of foods and beverages high in sugars is associated with an increased prevalence of obesity, insulin resistance, dyslipidemia and, more recently, ischemic heart and kidney diseases. The molecular mechanism(s) underlying the detrimental effects of sugar are not completely understood and their elucidation is critical to provide new insights on the health risk of fructose-based sweeteners. A better understanding of the key role of fructose overconsumption in the development of metabolic disorders may contribute to planning new strategies for preventing deleterious dietary behaviors from becoming established and, thus, curbing the rise in the number of insulin-resistant, obese and diabetic populations worldwide.

An environmental life cycle assessment comparing Australian sugarcane with US corn and UK sugar beet as producers of sugars for fermentation

Sugarcane is a highly suitable substrate for the production of bio-products. As well as producing high yields of sugar, much of the plant's fibre is also recovered and used as a source of renewable energy. A life cycle assessment (LCA) of sugarcane production and processing in Australia was performed to develop an environmental profile of sugarcane as a source of bio-products. The application examined was fermentation products from sugar. The sugarcane results were compared with results for other sugar producing crops—US corn and UK sugar beet—to gauge its relative environmental performance. The results show sugarcane to have an advantage in respect of energy input, greenhouse gas emissions and possibly acidification potential due to its high saccharide yield and the displacement of fossil fuels with surplus renewable energy from cane fibre (bagasse). However Australian sugarcane can exhibit high nitrous oxide emissions, which would reduce greenhouse gas advantages in some regions. For eutrophication, sugar beet provides advantages due to the avoided production of other agricultural crops displaced by the use of beet pulp as an animal feed. The three factors found to have the most influence on the environmental impacts of these agro-industrial systems were the commodities displaced by by-products, agricultural yields, and nitrogen use efficiency.

Quantification of anthocyanins in black carrot extracts (Daucus carota ssp. sativus var. atrorubens Alef.) and evaluation of their color properties

Pigment composition of 15 black carrot cultivars (Daucus carota ssp. sativus var. atrorubens Alef.) was screened by HPLC-MS. Up to seven cyanidin glycosides, five of which were acylated with hydroxycinnamic and hydroxybenzoic acids, were identified and quantified in the roots by HPLC-DAD. Contents of individual compounds indicated great differences in the potential of anthocyanin accumulation both between different cultivars and carrots of the same cultivar. Total anthocyanin amounts ranged from 45.4 mg/kg dry matter to 17.4 g/kg dry matter. To the best of our knowledge, this is the first report on the quantification of individual anthocyanins in roots of different black carrot cultivars. The determination of color properties in the extracts under various pH conditions proved black carrot anthocyanins to be applicable as natural food colorants also for low-acid food commodities, whereas a considerable loss of color was noted under nearly neutral conditions. Additionally, relatively high saccharide contents were found in almost all cultivars which may be disadvantageous when coloring concentrates are produced from carrot roots.

Reiterative Intramolecular Glycosylation Supported by a Rigid Spacer

This paper describes a synthesis of trisaccharide 1 by reiterative intramolecular glycosidation with 5-(bromomethyl)-2-methylbenzoic acid (5a) as starting material for the generation of a rigid spacer. Intramolecular glycosidation of donor−acceptor-tethered compound 24 yielded disaccharide 25. Transesterification of 25 afforded 26, which generated a new linking centre for the next spacer. Repetition of the previous cycle on 26 yielded trisaccharide 1, which again presents an extension point for the synthesis of higher saccharides. (© Wiley-VCH Verlag GmbH, 69451 Weinheim, Germany, 2003)

Glycosaminoglycan-mimetic biomaterials. 3. Glycopolymers prepared from alkene-derivatized mono- and disaccharide-based glycomonomers.

Mono- and disaccharide-containing glycopolymers were synthesized by two different free-radical processes, and their ability to act as heparan sulfate glycomimetics in promoting the binding of Fibroblast Growth Factor-2 (FGF-2) to its receptor (FGFR-1) was evaluated using an in vitro cell-based assay. Cyanoxyl (*OC triple bond N)-mediated polymerization of acrylamide with alkene-derivatized mono- and disaccharides including sulfated or nonsulfated N-acetyl-D-glucosamine is described. The results of this approach are compared to those obtained via the classical ammonium peroxodisulfate (APS)/N,N,N',N'-tetramethylethylenediamine (TMEDA) initiating system and confirm the capacity of cyanoxyl-mediated polymerization to generate a variety of glycopolymers with high saccharide contents and low polydispersity indexes. In vitro assays demonstrate that specific glycopolymers can potentiate FGF-2/FGFR-1 binding interactions.

Functional Improvements in Food Proteins in Multiple Aspects by Conjugation with Saccharides: Case Studies of .BETA.-Lactoglobulin-Acidic Polysaccharides Conjugates.

Many studies on the conjugates of proteins and saccharides (neoglycoproteins) have been carried out over the past 20 years to improve the function of proteins. However, most have been carried out by attachment or elimination of low-molecular weight molecules for single purposes. The structure-function relationship of the conjugates has not been well understood, and little is known about the improvement of proteins by conjugation with polysaccharides. We sought to achieve functional improvements in food proteins in multiple aspects by conjugation with charged polysaccharides. We chose bovine β-lactoglobulin (β-LG) as the target protein for improvement of the function and attached carboxymethyl dextrans (CMDs) of different molecular weight (10, 40, 70 and 162 kDa) covalently. After conjugation and purification, we explored the structure of the β-LG-CMD conjugates to prove that we could prepare conjugates while maintaining the native-like structure of protein molecule. By conjugation with CMDs, enhancement of thermal stability, improvement in the emulsifying properties and reduction of immunogenicity of β-LG could be achieved. Increases in the CMD content and net charge were viewed as the major factors in the improved emulsifying properties. Conjugation with high saccharide content using polysaccharides of higher molecular weight is believed to be effective in reduced immunogenicity of β-LG.