Rare Sugar Research Articles

Impact of D-allulose consumption on Enteric pathogens in human gut Microbiota: A randomized controlled trial study

D-allulose, a rare sugar recognized as Generally Recognized as Safe, has emerged as a potential alternative to sucrose. Despite its growing popularity, research on its effects on the human gut microbiota, including pathogens, remains scarce. To address these concerns, we conducted a 12-week randomized, double-blind, parallel, placebo-controlled study assessing D-allulose’s safety on gut microbiota in humans. Participants consumed 15 g/day of D-allulose or sucralose (placebo) for 12 weeks. Gut microbiota analysis in stool samples, performed through shotgun metagenomics sequencing before and after the intervention, evaluated microbial diversity, taxonomy of prevalent species, changes in pathogenic bacteria (Clostridium difficile, Helicobacter hepaticus, Klebsiella pneumoniae, Bacteroides fragilis, Staphylococcus aureus, and Salmonella enterica), and short-chain fatty acid production. Our findings revealed no significant differences in microbial diversity, pathogenic bacteria levels, or short-chain fatty acid production, suggesting that D-allulose consumption is safe and does not adversely affect the gut microbiome or pathogen presence.

A New Platform Molecule from Gluconolactone? Access to Furanics, Rare Sugars, β-Ketoamides and Amino Furanones.

Gluconolactone serves as a readily available and inexpensive starting material for synthesizing the potential platform chemical (Z)-3-deoxy-1,2:5,6-di-O-isopropylidene-D-erythro-hex-3-enolactone in two steps. In this work, the selective elimination of triacetone gluconolactone was optimized. The resulting product is a versatile molecule, capable of being transformed into various compound classes in one or a few steps, i.e. potentially a new biomass-based platform chemical. This study demonstrates how it can be transformed into furanics, rare sugars, β-ketoamides, and amino furanones.

Chemoenzymatic Synthesis of Phosphosaccharides from Haemophilus parasuis Strains ER-6P (Serovar 15) and Nagasaki (Serovar 5).

Herein, we describe a chemoenzymatic and diversity-oriented approach for the first syntheses of octasaccharide repeating units of the capsular polysaccharides of Haemophilus parasuis serovar 15 and serovar 5. The synthetic method features efficient enzymatic assembly of sialyl galactose or N-acetyl-galactosamine building blocks, highly stereoselective chemical construction of α-type H-phosphonate, and the β-stereospecific 1,3-glycosylation reaction of a rare sugar donor.

Epimerization of glucose to rare sugars using Beta zeolite-supported MoOx catalysts

Epimerization of abundant monosaccharides derived from lignocellulosic biomass offers an attractive approach for the synthesis of rare sugars. Molybdenum-based catalysts demonstrate excellent glucose-mannose epimerization performance, but most studies have neglected the synthesis of other sugar epimers and the pathways of their formation. Here, reduced Beta zeolite supported MoOx catalysts (Mo/Beta) with 1.5–10 wt% Mo loading were examined for glucose epimerization in water, specifically focusing on the formation of the rare sugars allose and altrose. The physicochemical structure and the catalytic activity of the catalyst were examined in detail, and the reaction mechanisms for the epimer formation were probed by nuclear magnetic resonance (NMR) spectroscopy. The results demonstrate that 5 wt% Mo/Beta achieved a near-equilibrium mannose yield of 30 % from glucose within 10 min at reaction temperatures below 140 °C, exhibiting a relatively low activation energy (∼67 kJ mol−1). At elevated reaction temperatures (≥ 120 °C), the rare sugars allose and altrose accumulated to combined yields of 24 % at 140 °C. An isotope labelling study using NMR spectroscopy corroborated the catalysis to involve 1,2-carbon shifts that elicit the formation of rare sugars at sufficiently high temperatures. Evaluation of catalyst reuse and regeneration indicated that supported MoOx had higher stability than MoO3, but the stability of the Mo/Beta catalysts under hydrothermal conditions leaves room for improvement.

Recent Progress in the Synthesis and Glycosylation of Rare Sugars

AbstractOut of 42 naturally occurring monosaccharides, only seven are abundant in Nature (glucose, galactose, mannose, fructose, xylose, ribose, and l-arabinose), while the others have been classified as ‘rare sugars’. Fungi and bacteria use a wide range of monosaccharides, in contrast to mammals, reflected in their glycosylated metabolites, as well as the cellular machineries that are involved in their sugar metabolism. Recognition of the microbiome’s impact on human health has led to increased interest in microbial glycans, as they often mediate interaction between host and microbes. Efficient access to rare sugars and oligosaccharides is necessary to study their roles in Nature, which can provide new pharmacological leads. Furthermore, it enables the synthesis of bioactive glycosylated natural products and congeners. This short review highlights recent progress in the synthesis and the efficient, site- and stereoselective glycosylation of rare sugars. Finally, it provides a recent example where synthetic access to rare sugars has enabled biochemical studies to better understand and interfere with processes in Nature.1 Introduction2 Synthesis of Rare Sugars2.1 Syntheses from Renewable Feedstock2.2 De Novo Syntheses3 Glycosylation3.1 Catalyst-Controlled Glycosylation3.2 One-Pot Iterative Oligosaccharide Synthesis in Solution4 Application in Biochemical Research5 Conclusion

Tailoring the natural rare sugars D-tagatose and L-sorbose to produce novel functional carbohydrates

This multidisciplinary study details the biosynthesis of novel non-digestible oligosaccharides derived from rare sugars, achieved through transfructosylation of D-tagatose and L-sorbose by levansucrase from Bacillus subtilis CECT 39 (SacB). The characterization of these carbohydrates using NMR and molecular docking was instrumental in elucidating the catalytic mechanism and substrate preference of SacB. Tagatose-based oligosaccharides were higher in abundance than L-sorbose-based oligosaccharides, with the most representative structures being: β-D-Fru-(2→6)-β-D-Fru-(2→1)-D-Tag and β-D-Fru-(2→1)-D-Tag. In vitro studies demonstrated the resistance of tagatose-based oligosaccharides to intestinal digestion and their prebiotic properties, providing insights into their structure-function relationship. β-D-Fru-(2→1)-D-Tag was the most resistant structure to small-intestinal digestion after three hours (99.8% remained unaltered). This disaccharide and the commercial FOS clustered in similar branches, indicating comparable modulatory properties on human fecal microbiota, and exerted a higher bifidogenic effect than unmodified tagatose. The bioconversion of selected rare sugars into β-fructosylated species with a higher degree of polymerization emerges as an efficient strategy to enhance the bioavailability of these carbohydrates and promote their interaction with the gut microbiota. These findings open up new opportunities for tailoring natural rare sugars, like D-tagatose and L-sorbose, to produce novel biosynthesized carbohydrates with functional and structural properties desirable for use as emerging prebiotics and low-calorie sweeteners.

Functionalized mesoporous biosilica for immobilizing D-allulose 3-epimerase: A multienzyme cascade strategy for calorie reduction in fruit tea drinks

D-allulose, a low-calorie rare sugar, can be biosynthesized by D-allulose 3-epimerase (DAE). Here, DAE was covalently immobilized on polyethyleneimine-modified diatomite using two methods: a biopolymer coating approach (DAE@PEI-GA@PDP) and a direct attachment method (DAE-GA@PEI@PDP). Both immobilized DAE enzymes exhibited enhanced thermal stability and broader pH tolerance compared to the free DAE. They also exhibited excellent reusability and maintained more than 70% of their activity after 10 cycles. Additionally, a multienzyme cascade system was developed to produce D-allulose from low-cost D-glucose using immobilized glucose isomerase (GI) and immobilized DAE enzymes. This system obtained 76 g/L D-allulose from 500 g/L D-glucose. Notably, the application was extended to fruit tea drinks, successfully transforming high-calorie sugars (D-glucose and D-fructose) into D-allulose. This research offers a promising immobilization strategy for DAE on silica-based materials and provides robust biocatalysts for creating functional fruit tea drinks aimed at diabetics and individuals with specific dietary needs.

Biological Activities of Soy Protein Hydrolysate Conjugated with Mannose and Allulose.

The non-enzymatic conjugation of peptides through the Maillard reaction has gained attention as an effective method to enhance biological functions. This study focuses on two conjugate mixtures: crude soy protein hydrolysate (SPH) conjugated with mannose (SPHM) and crude soy protein hydrolysate conjugated with allulose (SPHA). These two mixtures were products of the Maillard reaction, also known as non-enzymatic glycation. In vitro experiments were conducted to evaluate the antioxidant, anti-pancreatic lipase, inhibition of Bovine Serum Albumin (BSA) denaturation, and anti-angiotensin converting enzyme (ACE) activities of these conjugated mixtures. The results indicate that conjugated mixtures significantly enhance the antioxidant potential demonstrated via the DPPH and FRAP assays. SPHA exhibits superior DPPH scavenging activity (280.87 ± 16.39 µg Trolox/mL) and FRAP value (38.91 ± 0.02 mg Trolox/mL). Additionally, both conjugate mixtures, at a concentration of 10 mg/mL, enhance the BSA denaturation properties, with SPHM showing slightly higher effectiveness compared to SPHA (19.78 ± 2.26% and 5.95 ± 3.89%, respectively). SPHA also shows an improvement in pancreatic lipase inhibition (29.43 ± 1.94%) when compared to the SPHM (23.34 ± 3.75%). Furthermore, both the conjugated mixtures and rare sugars exhibit ACE inhibitory properties on their own, effectively reducing ACE activity. Notably, the ACE inhibitory effects of the individual compounds and their conjugate mixtures (SPHM and SPHA) are comparable to those of positive control (Enalapril). In conclusion, SPHM and SPHA demonstrate a variety of bioactive properties, suggesting their potential use in functional foods or as ingredients in supplementary products.

Chronic Use of Artificial Sweeteners: Pros and Cons.

Over the past few decades, the scientific community has been highly concerned about the obesity epidemic. Artificial sweeteners are compounds that mimic the sweet taste of sugar but have no calories or carbohydrates; hence, they are very popular among patients suffering from diabetes or obesity, aiming to achieve glycemic and/or weight control. There are four different types of sweeteners: artificial, natural, rare sugars, and polyols. Artificial and natural sweeteners are characterized as non-nutritional sweeteners (NNSs) since they do not contain calories. The extended use of sweeteners has been reported to have a favorable impact on body weight and glycemic control in patients with type 2 diabetes (T2DM) and on tooth decay prevention. However, there is concern regarding their side effects. Several studies have associated artificial sweeteners' consumption with the development of insulin resistance, nonalcoholic fatty liver disease (NAFLD), gastrointestinal symptoms, and certain types of cancer. The present review focuses on the description of different types of sweeteners and the benefits and possible deleterious effects of the chronic consumption of NNSs on children's health. Additionally, possible underlying mechanisms of the unfavorable effects of NNSs on human health are described.

Total synthesis of a structurally complex zwitterionic hexasaccharide repeating unit of polysaccharide B from Bacteroides fragilis via one-pot glycosylation

Zwitterionic polysaccharides (ZPSs) present on the surface of a common gut commensal Bacteroides fragilis are endowed with unique immunological properties as they can directly bind to T-cells in the absence of protein conjugation. ZPSs are therefore considered to be potential antigens for the development of totally carbohydrate-based vaccines. Herein, we disclose the first total synthesis of a highly branched phosphorylated zwitterionic capsular polysaccharide repeating unit of Bacteroides fragilis. The hexasaccharide repeating unit bearing six different monosaccharides comprises three 1,2-cis-glycosidic linkages, a challenging 1,2-trans linkage in D-QuipNAc-β-(1→4)-D-Gal motif, and a 2-aminoethyl phosphonate appendage. The synthesis of target ZPS was accomplished utilizing an expeditious, highly stereoselective and convergent (1 + 2 + 2 + 1) one-pot glycosylation strategy. The striking features include efficient synthesis of rare deoxy amino sugars D- and L-quinovosamine, stereoselective installation of three 1,2-cis glycosidic linkages, glycosylation of D-quinovosamine donor with a sterically crowded, poorly reactive 4-OH galactose moiety, as well as late stage phosphorylation.

Low-calorie d-allulose as a sucrose alternative modulates the physicochemical properties and volatile profile of sponge cake.

d-Allulose, a C-3 epimer of d-fructose, is a rare sugar with ∼70% of the sweetness of sucrose but a caloric content of only 0.4kcal/g. Due to its low-calorie nature, d-allulose has garnered increasing interest in the food industry. This study was the first attempt to explore the effect of d-allulose as a sucrose replacer on the properties of sponge cake, a widely consumed high-sugar product. Substituting sucrose with d-allulose generated negligible impact on the batter system, while pronounced differences in physicochemical properties of cakes were detected, including specific volume, texture, microstructure, color, and antioxidant activity. In addition, sponge cake containing d-allulose displayed a distinctive aroma volatile profile, with more furans and pyrazines generation. Furthermore, correlations of physicochemical properties across all formulations were depicted, and the potential mechanism behind the property alterations modulated by d-allulose was revealed from the perspectives of starch gelatinization and browning reactions. Overall, this study provides insights into the application potential of d-allulose as a sucrose substitute in bakery product. PRACTICAL APPLICATION: This study elucidates the effect of d-allulose as a low-calorie sugar substitute on sponge cakes. This finding is valuable for the food industry, providing insights into a healthier alternative to traditional sugar in baked goods.

Effective catalytic conversion of cellulose pyrolysis into D-Allose via Al1-Fe5 nano-catalysts

This research has made significant advancements in the field of high-value bio-oil production, specifically focusing on the synthesis of the rare sugar D-Allose. Notably, synthesizing D-Allose through cellulose pyrolysis has long been challenging due to its high synthetic difficulty, which has limited related studies. By employing a one-pot synthesis approach using Alx-Fey nano-catalysts prepared through a unique cellulose pretreatment method, this experiment successfully achieved efficient generation of D-Allose during rapid pyrolysis. Systematic experimentation demonstrated that at 330 °C with the Al1-Fe5 nano-catalyst, the yield of D-Allose from cellulose pyrolysis reached an impressive 50.69 %, accompanied by a detection of 14.35 % furfural in the bio-oil. This breakthrough not only significantly contributes to the high-value utilization of bio-oil but also provides robust support for further development in this area. Simultaneously, the success of this research extends beyond high-value bio-oil utilization and makes a substantial impact on both the scientific community and industry.

Development and Characterization of a Tunable Metal-Organic Framework (MOF) for the Synthesis of a Rare Sugar D-Tagatose.

D-tagatose is a valuable rare sugar with potential health benefits such as antiobesity, low-calorie, prebiotic, and anticancer. However, its production is mainly depending on chemical or enzymatic catalysis. Herein, a cobalt-based metal-organic framework (MOF) was developed at room temperature in an aqueous system using a self-assembly method. The L-arabinose isomerase (L-AI) was immobilized into this unique MOF by an in situ encapsulation process. The morphology and structural aspects of the MOF preparations were characterized by different analytical techniques such as scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), Fourier transform infrared spectroscopy (FT-IR), and X-Ray diffraction (XRD). Moreover, thermogravimetric analysis (TGA) suggested the high thermal stability of the L-AI@MOF. Significantly, the immobilized catalyst exhibited enhanced catalytic efficiency (kcat/Km) of 3.22mM-1s-1 and improved turnover number (kcat) of 57.32s-1. The L-AI@MOF efficiently catalyzes the synthesis of D-tagatose from D-galactose up to the equilibrium level (~ 50%) of isomerization in heterogeneous catalysis. Interestingly, L-AI@MOF was found stable and reusable for more than five cycles without the requirement of additional metal ions during catalysis. Thus, L-AI stabilized in the MOF system demonstrated a higher catalytic activity and potential guidance for the sustainable synthesis of rare sugar D-tagatose.

Reshaping the binding pocket of D-tagaturonate epimerase UxaE to improve the epimerization activity of C4-OH for enabling green synthesis of d-tagatose

d-tagatose, a rare sugar with broad applications in the food and pharmaceutical industries, can be synthesized from d-fructose by epimerases, an approach with significant biosynthesis potential. However, so far there has been only one attempt to engineer tagatose 4‑epimerase through random mutagenesis. The lack of exploration has severely restricted the application of epimerases in the industrial production of d-tagatose. The previous research primarily focused on enhancing the catalytic activity of epimerases through random mutagenesis, with limited understanding of how substrate-adjacent residues affect enzyme efficiency, which was hence the subject of this study. First, using a semi-rational design approach, after three rounds of mutagenesis and screening, the mutant (S125D/S268A/Q124E) of D-tagaturonate epimerase (UxaE) from Thermotoga neapolitana was obtained, which increased d-tagatose production efficiency by 10.8-fold compared to the wild type, with a specific activity of 1505.6 U/g, representing the highest specific activity reported for a UxaE mutant to date. Molecular dynamics simulations showed that the substitution of Ser to Asp-125 enhanced substrate-enzyme interactions, while the substitution of Glu to Gln-124 reinforced the hydrogen bond network around the catalytic residue Glu-128, further reshaping the substrate-binding pocket and enhancing catalytic activity. This study for the first time explored the enhancement of the C4 epimerization function of epimerases by reshaping their substrate-binding pockets, thereby providing a feasible route for the industrial biosynthesis of d-tagatose.

Stereoselective Construction of Multifunctional C-Glycosides Enabled by Nickel-Catalyzed Tandem Borylation/Glycosylation.

Stereochemically pure saccharides have indispensable roles in fields ranging from medicinal chemistry to materials science and organic synthesis. However, the development of a simple, stereoselective, and efficient glycosylation protocol to access α- and β-C-glycosides (particularly 2-deoxy entities) remains a persistent challenge. Existing studies have primarily focused on C1 modification of carbohydrates and transformation of glycosyl radical precursors. Here, we innovate by harnessing the in situ generated glycosyl-Ni species to achieve one-pot borylation and glycosylation in a cascade manner, which is enabled by an earth-abundant nickel-catalyzed carboboration of readily accessible glycals without any ligand. This work reveals the potential for the development of a modular and multifunctional glycosylation platform to facilitate the simultaneous introduction of C-C and C-B bonds at the stereogenic center of saccharides, a largely unexploited research area. Preliminary experimental and computational studies indicate that the endocyclic O and the C3 group play important roles in stereoseclectively forging glycosidic bonds. As a result, a diverse range of C-R (R = alkyl, aryl, and alkenyl) and 2-deoxygenated glycosides bearing modifiable boron groups could be rapidly made with excellent stereocontrol and exhibit remarkable functional group tolerance. The synthetic potential is underscored in the late-stage glycosylation of natural products and commercial drugs as well as the facile preparation of various rare sugars, bioactive conjugates, and key intermediates to prorocentin, phomonol, and aspergillide A.

In Silico Analysis and Development of the Secretory Expression of D-Psicose-3-Epimerase in Escherichia coli.

D-psicose-3-epimerase (DPEase), a key enzyme for D-psicose production, has been successfully expressed in Escherichia coli with high yield. However, intracellular expression results in high downstream processing costs and greater risk of lipopolysaccharide (LPS) contamination during cell disruption. The secretory expression of DPEase could minimize the number of purification steps and prevent LPS contamination, but achieving the secretion expression of DPEase in E. coli is challenging and has not been reported due to certain limitations. This study addresses these challenges by enhancing the secretion of DPEase in E. coli through computational predictions and structural analyses. Signal peptide prediction identified PelB as the most effective signal peptide for DPEase localization and enhanced solubility. Supplementary strategies included the addition of 0.1% (v/v) Triton X-100 to promote protein secretion, resulting in higher extracellular DPEase (0.5 unit/mL). Low-temperature expression (20 °C) mitigated the formation of inclusion bodies, thus enhancing DPEase solubility. Our findings highlight the pivotal role of signal peptide selection in modulating DPEase solubility and activity, offering valuable insights for protein expression and secretion studies, especially for rare sugar production. Ongoing exploration of alternative signal peptides and refinement of secretion strategies promise further enhancement in enzyme secretion efficiency and process safety, paving the way for broader applications in biotechnology.

Production, purification, characterization, and safety evaluation of constructed recombinant D-psicose 3-epimerase

BackgroundD-psicose 3-epimerase (DPEase) is a potential catalytic enzyme for D-psicose production. D-psicose, also known as D-allulose, is a low-calorie sweetener that has gained considerable attention as a healthy alternative sweetener due to its notable physicochemical properties. This research focused on an in-depth investigation of the expression of the constructed DPEase gene from Agrobacterium tumefaciens in Escherichia coli for D-psicose synthesis. Experimentally, this research created the recombinant enzyme, explored the optimization of gene expression systems and protein purification strategies, investigated the enzymatic characterization, and then optimized the D-psicose production. Finally, the produced D-psicose syrup underwent acute toxicity evaluation to provide scientific evidence supporting its safety.ResultsThe optimization of DPEase expression involved the utilization of Mn2+ as a cofactor, fine-tuning isopropyl β-D-1-thiogalactopyranoside induction, and controlling the induction temperature. The purification process was strategically designed by a nickel column and an elution buffer containing 200 mM imidazole, resulting in purified DPEase with a notable 21.03-fold increase in specific activity compared to the crude extract. The optimum D-psicose conversion conditions were at pH 7.5 and 55 °C with a final concentration of 10 mM Mn2+ addition using purified DPEase to achieve the highest D-psicose concentration of 5.60% (w/v) using 25% (w/v) of fructose concentration with a conversion rate of 22.42%. Kinetic parameters of the purified DPEase were Vmax and Km values of 28.01 mM/min and 110 mM, respectively, which demonstrated the high substrate affinity and efficiency of DPEase conversion by the binding site of the fructose-DPEase-Mn2+ structure. Strategies for maintaining stability of DPEase activity were glycerol addition and storage at -20 °C. Based on the results from the acute toxicity study, there was no toxicity to rats, supporting the safety of the mixed D-fructose–D-psicose syrup produced using recombinant DPEase.ConclusionsThese findings have direct and practical implications for the industrial-scale production of D-psicose, a valuable rare sugar with a broad range of applications in the food and pharmaceutical industries. This research should advance the understanding of DPEase biocatalysis and offers a roadmap for the successful scale-up production of rare sugars, opening new avenues for their utilization in various industrial processes.

Research progress in physiological function of the rare sugar D-allose

D-allose, a rare sugar with anti-oxidant, anti-inflammatory, anti-cancer, immunosuppressing and other physiological functions, has become a research hotspot in recent years. This paper describes the physical and chemical properties, synthesis methods, metabolism, physiological functions, and applications of D-allose, aiming to promote the functional development of D-allose and facilitate the application of D-allose in the food field and clinical treatment.

The Effect of Allulose on the Attenuation of Glucose Release from Rice in a Static In Vitro Digestion Model.

Allulose is a rare sugar that provides <10% of the energy but 70% of the sweetness of sucrose. Allulose has been shown to attenuate glycemic responses to carbohydrate-containing foods in vivo. This study aimed to determine the optimal allulose dose for minimizing in vitro glucose release from rice compared to a rice control and fructose. A triphasic static in vitro digestion method was used to evaluate the in vitro digestion of a rice control compared to the co-digestion of rice with allulose (10 g, 20 g, and 40 g) and fructose (40 g). In vitro glucose release was affected by treatment (p < 0.001), time (p < 0.001), and treatment-by-time interaction (p = 0.002). Allulose (40 g) resulted in a reduction in in vitro glucose release from rice alone and rice digested with allulose (10 g), allulose (20 g), and fructose. The incremental area under the curve (iAUC) for in vitro glucose release was lower after allulose (40 g) (p = 0.005) compared to rice control and allulose (10 g) but did not differ from allulose (20 g) or fructose. This study demonstrates that allulose reduces glucose release from carbohydrates, particularly at higher doses, underscoring its potential as a food ingredient with functional benefits.

Impact of Rare Sugar D-Allulose on Hardening of Starch Gels during Refrigerated Storage.

The rare sugar D-allulose (Alu), with ca. 10% calories of sucrose (Suc), is a promising alternative sugar that can be used to improve the quality of starch gels in storage. The effects of Alu (compared to Suc) on the hardening and microstructural and molecular order of amylopectin-rich (glutinous rice (GR) and corn amylopectin (CAP)) and amylose-rich (corn (C)) starch gels were investigated. Alu and Suc both suppressed hardening in C gels, while Alu but not Suc was effective in GR and CAP gels. SEM results showed that Alu-containing GR and CAP maintained a relatively large pore size compared to Suc-containing gels. The deconvolution of FTIR spectra revealed that Alu-containing GR and CAP gels had lower ratios of intermolecular hydrogen bonds and higher ratios of loose hydrogen bonds than Suc-containing gels. For amylose-rich C gels, on the other hand, such tendencies were not observed. The influence of Alu on amylopectin-rich gels could be because Alu reduced the ratio of intermolecular hydrogen bonds, which might be involved in amylopectin recrystallization, and increased that of loose hydrogen bonds. The results suggest that Alu is more effective than Suc in inhibiting the hardening of amylopectin-rich starch gels during refrigerated storage.