Search

Isomaltulose is an ideal functional sweetener and has been approved as a safe sucrose substitute. It is produced mainly through sucrose isomerization catalyzed by sucrose isomerase. Here, to produce food-grade isomaltulose and improve its yield, the sucrose isomerase gene from Pantoea dispersa UQ68J was overexpressed in the non-pathogenic yeast Yarrowia lipolytica. When the engineered strain, S47, was fermented on 600 g/L sucrose in a 10-L bioreactor, a maximum isomaltulose concentration of 572.1 g/L was achieved. Sucrose isomerase activity was 7.43 U/mL, and yield reached 0.96 g/g. Moreover, monosaccharide byproducts were simultaneously transformed into intracellular lipids, thus reducing the production of undesirable compounds and resulting in high isomaltulose purity (97.8%) in the final broth. In summary, the bioprocess employed in this study provides an efficient alternative strategy for isomaltulose production., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Bifidobacteria are a major component of the intestinal microbiota in humans, particularly breast-fed infants. Therefore, elucidation of the mechanisms by which these bacteria colonize the intestine is desired. One approach is transposon mutagenesis, a technique currently attracting much attention because, in combination with next-generation sequencing, it enables exhaustive identification of genes that contribute to microbial fitness. We now describe a transposon mutagenesis system for Bifidobacterium longum subsp. longum 105-A (JCM 31944) based on IS Blo11 , a native IS 3 family insertion sequence. To build this system, xylose-inducible or constitutive bifidobacterial promoters were tested to drive the expression of full-length or a truncated form at the N terminus of the IS Blo11 transposase. An artificial transposon plasmid, pBFS12, in which IS Blo11 terminal inverted repeats are separated by a 3-bp spacer, was also constructed to mimic the transposition intermediate of IS 3 elements. The introduction of this plasmid into a strain expressing transposase resulted in the insertion of the plasmid with an efficiency of >10 3 CFU/μg DNA. The plasmid targets random 3- to 4-bp sequences, but with a preference for noncoding regions. This mutagenesis system also worked at least in B. longum NCC2705. Characterization of a transposon insertion mutant revealed that a putative α-glucosidase mediates palatinose and trehalose assimilation, demonstrating the suitability of transposon mutagenesis for loss-of-function analysis. We anticipate that this approach will accelerate functional genomic studies of B. longum subsp. longum IMPORTANCE Several hundred species of bacteria colonize the mammalian intestine. However, the genes that enable such bacteria to colonize and thrive in the intestine remain largely unexplored. Transposon mutagenesis, combined with next-generation sequencing, is a promising tool to comprehensively identify these genes but has so far been applied only to a small number of intestinal bacterial species. In this study, a transposon mutagenesis system was established for Bifidobacterium longum subsp. longum , a representative health-promoting Bifidobacterium species. The system enables the identification of genes that promote colonization and survival in the intestine and should help illuminate the physiology of this species., (Copyright © 2018 American Society for Microbiology.)

Insulin resistance is associated with a poor prognosis in non‑alcoholic fatty liver disease (NAFLD) patients. Isomaltulose, a naturally‑occurring disaccharide, is reported to improve glucose and lipid metabolisms in obese patients. The present study aimed to investigate the effects of isomaltulose on insulin resistance and various metabolites in NAFLD patients. Five male patients with NAFLD consumed 20 g isomaltulose or sucrose (control). Changes in insulin resistance and metabolites were evaluated by alterations of serum C‑peptide immunoreactivity (CPR) and metabolomic analysis from baseline to 15 min after the administration, respectively. There was no significant difference in changes of blood glucose level; however, the CPR level was significantly decreased in the Isomaltulose group compared to the control group (0.94±0.89 vs. ‑0.12±0.31, P=0.0216). In a metabolomic analysis, a significant alteration was seen in 52 metabolites between the control and Isomaltulose groups. In particular, the taurodeoxycholic acid level significantly increased approximately 12.5‑fold, and the arachidonic acid level significantly decreased approximately 0.01‑fold. Together, it present study demonstrated that isomaltulose improved insulin resistance in NAFLD patients. It was also revealed that isomaltulose affects taurodeoxycholic acid and arachidonic acid. Thus, isomaltulose may have a beneficial effect on insulin resistance through alterations of bile acid and fatty acid metabolisms in NAFLD patients.

Purpose: The performance and physiological effects of isomaltulose and maltodextrin consumed intermittently during prolonged soccer-specific exercise were investigated., Methods: University soccer players (n = 22) performed 120 min of intermittent exercise while consuming 8% carbohydrate-electrolyte drinks (equivalent to ~ 20 g h -1 ) containing maltodextrin (Glycaemic Index: 90-100), isomaltulose (Glycaemic Index: 32) or a carbohydrate-energy-free placebo in a manner replicating the practices of soccer players (i.e., during warm-up and half-time). Physical (sprinting, jumping) and technical (shooting, dribbling) performance was assessed., Results: Blood glucose and plasma insulin (both P < 0.001) concentrations varied by trial with isomaltulose maintaining > 13% higher blood glucose concentrations between 75 and 90 min versus maltodextrin (P < 0.05). A decline in glycaemia at 60 min in maltodextrin was attenuated with isomaltulose (-19 versus -4%; P = 0.015). Carbohydrates attenuated elevations in plasma epinephrine concentrations (P < 0.05), but isomaltulose proved most effective at 90 and 120 min. Carbohydrates did not attenuate IL-6 increases or reductions in physical or technical performances (all P > 0.05). Ratings of abdominal discomfort were influenced by trial (P < 0.05) with lower values for both carbohydrates compared to PLA from 60 min onwards., Conclusions: Although carbohydrates (~ 20 g h -1 ) did not attenuate performance reductions throughout prolonged soccer-specific exercise, isomaltulose maintained higher blood glucose at 75-90 min, lessened the magnitude of the exercise-induced rebound glycaemic response and attenuated epinephrine increases whilst maintaining similar abdominal discomfort values relative to maltodextrin. When limited opportunities exist to consume carbohydrates on competition-day, low-glycaemic isomaltulose may offer an alternative nutritional strategy for exercising soccer players.

A green process and environmentally benign process is highly desirable in the development of enzymatic catalysis. In this work, the shuttle plasmid pHA01 was constructed and the sucrose isomerase (SIase) was expressed in Bacillus subtilis WB800. The optimal nitrogen and carbon sources for SIase expression were yeast extract (15g/L) and un-pretreated cane molasses (UCM, 20g/L), respectively. After the UCM fed, the whole cell activity reached 5.2U/mL in a 7.5L fermentor. Optimum catalytic temperature and pH of whole cell were 35°C and 5.5, respectively. Although the biologic membrane reactor (BMR) system consecutively worked for 12 batches, the sucrose conversion remained higher than 90%, indicating the BMR system had a greater operational stability. Furthermore, isomaltulose production using the BMR system with low-cost cane molasses as its substrate not only reduces the production cost and mediates environmental pollution, but also solves the genetic background problem of the non-food-grade strains., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Low glycaemic index (GI) foods minimize large blood glucose fluctuations and have been advocated to enhance fat oxidation and may contribute to weight management. We determined whether the inclusion of isomaltulose compared to sucrose in a low/high GI meal sequence can modulate the glycaemic response and substrate oxidation in an Asian population. Twenty Chinese men (body mass index (BMI): 17-28 kg/m²) followed a 24 h low GI (isomaltulose, Palatinose TM ) or high GI (sucrose) diet in a randomized double-blind, controlled cross-over design. Treatment meals included dinner (day 1), breakfast, lunch, and snack (day 2). Continuous glucose monitoring provided incremental area under the curve (iAUC) and mean amplitude of glycaemic excursion (MAGE) and 10 h indirect calorimetry (whole body calorimeter) (day 2) provided energy expenditure and substrate oxidation. Our results demonstrated that the low GI diet resulted in lower 24 h glucose iAUC (502.5 ± 231.4 vs. 872.6 ± 493.1 mmol/L; p = 0.002) and lower 24 h glycaemic variability (MAGE: 1.67 ± 0.53 vs. 2.68 ± 1.13 mmol/L; p < 0.001). Simultaneously, 10 h respiratory quotient increased more during high GI ( p = 0.014) and fat oxidation was higher after low GI breakfast ( p = 0.026), lunch ( p < 0.001) and snack ( p = 0.013). This indicates that lower GI mixed meals incorporating isomaltulose are able to acutely reduce the glycaemic response and variability and promote fat oxidation.

Isomaltulose, a naturally-occurring isomer of sucrose, is commonly used as an alternative sweetener in foods and beverages. The goal of this study was to determine the effect of isomaltulose together with green tea on postprandial plasma glucose and insulin concentration, as well as antioxidant capacity in healthy subjects. In a randomized, single-blind, crossover study, 15 healthy subjects (eight women and seven men; ages 23.5 ± 0.7 years; with body mass index of 22.6 ± 0.4 kg/m²) consumed five beverages: (1) 50 g sucrose in 400 mL water; (2) 50 g isomaltulose in 400 mL of water; (3) 400 mL of green tea; (4) 50 g sucrose in 400 mL of green tea; and (5) 50 g isomaltulose in 400 mL of green tea. Incremental area under postprandial plasma glucose, insulin, ferric reducing ability of plasma (FRAP) and malondialdehyde (MDA) concentration were determined during 120 min of administration. Following the consumption of isomaltulose, the incremental 2-h area under the curve (AUC 0-2 h ) indicated a higher reduction of postprandial glucose (43.4%) and insulin concentration (42.0%) than the consumption of sucrose. The addition of green tea to isomaltulose produced a greater suppression of postprandial plasma glucose (20.9%) and insulin concentration (37.7%). In accordance with antioxidant capacity, consumption of sucrose (40.0%) and isomaltulose (28.7%) caused the reduction of green tea-induced postprandial increases in FRAP. A reduction in postprandial MDA after drinking green tea was attenuated when consumed with sucrose (34.7%) and isomaltulose (17.2%). In conclusion, green tea could enhance the reduction of postprandial glucose and insulin concentration when consumed with isomaltulose. In comparison with sucrose, isomaltulose demonstrated less alteration of plasma antioxidant capacity after being consumed with green tea.

Low glycemic index diets are supposed to achieve a more beneficial effect on blood glucose control in people with diabetes mellitus and may also provide metabolic benefits for the general population. A prototype of a low-glycemic index carbohydrate is the natural occurring disaccharide isomaltulose that can be commercially produced from sucrose (beet sugar) to industrial scale. It is currently used in various food and drink applications as well as special and clinical nutrition feeds and formula diet as a food ingredient and alternative sugar. Here we provide an overview on clinical trials with isomaltulose including an analysis of its effects on glycemia and fat oxidation as compared to high glycemic index sugars and carbohydrates. In addition, we discuss recent reports on beneficial effects in weight-loss maintenance and pregnancy.

The aim of this study was to compare the glycemic response of Caucasians and Asians to two disaccharides of different glycemic index (GI), and to examine if ethnic groups that showed the largest glycemic response to sucrose would benefit the most when it is replaced with isomaltulose. Forty healthy participants (10 Chinese; 10 Malays; 10 Caucasians; and 10 Indians) consumed beverages containing 50 g of sucrose or isomaltulose on two separate occasions using a randomized crossover design. Capillary blood glucose was measured in a fasted state and at 15, 30, 45, 60, 90, and 120 min after beverage ingestion. Glycemic response to sucrose was significantly higher in Malays compared to Caucasians ( p = 0.041), but did not differ between Caucasians vs. Chinese ( p = 0.145) or vs. Indians ( p = 0.661). When sucrose was replaced with isomaltulose, glycemic responses were significantly reduced in all ethnic groups, with the largest reduction in glycemic response being observed in Malays. Malays, who had the greatest glycemic response to sucrose, also showed the greatest improvement in glycemic response when sucrose was replaced with isomaltulose. This implies that Malays who are more susceptible to type 2 diabetes mellitus may benefit from strategies that replace high GI carbohydrate with lower GI alternatives to assist in glycemic control.

Among members of the glycoside hydrolase (GH) family, sucrose isomerase (SIase) and oligo-1,6-glucosidase (O16G) are evolutionarily closely related even though their activities show different specificities. A gene (Avin&#95;08330) encoding a putative SIase (AZOG: Azotobacterglucocosidase) from the nitrogen-fixing bacterium Azotobacter vinelandii is a type of pseudo-SIase harboring the "RLDRD" motif, a SIase-specific region in 329-333. However, neither sucrose isomerization nor hydrolysis activities were observed in recombinant AZOG (rAZOG). The rAZOG showed similar substrate specificity to Bacillus O16G as it catalyzes the hydrolysis of isomaltulose and isomaltose, which contain α-1,6-glycosidic linkages. Interestingly, rAZOG could generate isomaltose from the small substrate methyl-α-glucoside (MαG) via intermolecular transglycosylation. In addition, sucrose isomers isomaltulose and trehalulose were produced when 250 mM fructose was added to the MαG reaction mixture. The conserved regions I and II of AZOG are shared with many O16Gs, while regions III and IV are very similar to those of SIases. Strikingly, a shuffled AZOG, in which the N-terminal region of SIase containing conserved regions I and II was exchanged with the original enzyme, exhibited a production of sucrose isomers. This study demonstrates an evolutionary relationship between SIase and O16G and suggests some of the main regions that determine the specificity of SIase and O16G., (Copyright © 2016 Elsevier Inc. All rights reserved.)

This study assessed the effect of carbohydrate intake on self-selected soccer-specific running performance. Sixteen male soccer players (age 23 ± 4 years; body mass 76.9 ± 7.2 kg; predicted VO 2max = 54.2 ± 2.9 mL∙kg -1 ∙min -1 ; soccer experience 13 ± 4 years) completed a progressive multistage fitness test, familiarisation trial and two experimental trials, involving a modified version of the Loughborough Intermittent Shuttle Test (LIST) to simulate a soccer match in a fed state. Subjects completed six 15 min blocks (two halves of 45 min) of intermittent shuttle running, with a 15-min half-time. Blocks 3 and 6, allowed self-selection of running speeds and sprint times, were assessed throughout. Subjects consumed 250 mL of either a 12% carbohydrate solution (CHO) or a non-caloric taste matched placebo (PLA) before and at half-time of the LIST. Sprint times were not different between trials (CHO 2.71 ± 0.15 s, PLA 2.70 ± 0.14 s; p = 0.202). Total distance covered in self-selected blocks (block 3: CHO 2.07 ± 0.06 km; PLA 2.09 ± 0.08 km; block 6: CHO 2.04 ± 0.09 km; PLA 2.06 ± 0.08 km; p = 0.122) was not different between trials. There was no difference between trials for distance covered ( p ≥ 0.297) or mean speed ( p ≥ 0.172) for jogging or cruising. Blood glucose concentration was greater ( p < 0.001) at the end of half-time during the CHO trial. In conclusion, consumption of 250 mL of 12% CHO solution before and at half-time of a simulated soccer match does not affect self-selected running or sprint performance in a fed state., Competing Interests: Ian Rollo is an employee of the Gatorade Sports Science Institute, a division of PepsiCo Inc. The views expressed in this manuscript are those of the authors and do not necessarily reflect the position or policy of PepsiCo Inc.

Intake of sugar-sweetened beverages (SSB) may contribute to cardiovascular risk. The aim of this study was to investigate whether functional sugars with low compared to high glycemic index (GI) have beneficial effects on arterial stiffness during a period of low-physical activity. In a controlled cross-over dietary intervention (55% CHO, 30% fat, 15% protein), 13 healthy men (age: 23.7 ± 2.2 years, body mass index: 23.6 ± 1.9 kg/m²) completed 2 × 1 week of low physical activity following 1 week of normal physical activity (2363 ± 900 vs. 11,375 ± 3124 steps/day). During inactive phases participants consumed either low-GI (isomaltulose) or high-GI SSB (maltodextrin-sucrose), providing 20% of energy requirements. Postprandial vasodilatation (augmentation index, AIx), insulin sensitivity (IS) and Glucagon-like-peptide 1 (GLP-1) responses were measured during a meal test before and after SSB-intervention. Compared to maltodextrin-sucrose-SSB, postprandial vasodilatation was prolonged (AIx after 120 min: 9.9% ± 4.3% vs. 11.4% ± 3.7%, p < 0.05) and GLP-1 secretion was higher with isomaltulose-SSB (total area under the GLP-1 curve (tAUC GLP )-1: 8.0 ± 4.4 vs. 5.4 ± 3.4 pM × 3 h; p < 0.05). One week of low-physical activity led to impaired IS that was attenuated with low-GI SSB consumption, but did not affect arterial stiffness ( p > 0.05). Higher postprandial GLP-1 secretion after intake of low compared to high-GI beverages may contribute to improved postprandial vasodilatation. Although one week of low-physical activity led to marked impairment in IS, it had no effect on arterial stiffness in healthy men., Competing Interests: The authors declare no conflict of interest.

Reducing the glycemic load (GL) of the diet may benefit appetite control but its utility is complicated by psychological influences on eating. Disinhibited behaviour, a risk factor for overconsumption, is characterized by reduced prefrontal cortex activity, which in turn modulates vagal tone; a phenomenon associated with glucoregulation. This double blind randomised controlled trial explored for the first time the influence of disinhibited eating and vagal tone (heart rate variability (HRV)) on hunger and the postprandial response to GL. Blood glucose (BG) and hunger were measured 30 and 150 min after consumption of water, glucose or isomaltulose (low glycemic sugar). After consuming glucose, independently of BMI or habitual diet, those with the highest levels of disinhibition had higher BG levels after thirty minutes (B = 0.192, 95% CI LL. 086, UL 0.297), and lower BG after one hundred and fifty minutes (B = -0.240, 95% CI LL -0.348, UL -0.131). BG was related to hunger but only in low disinhibited eaters. Disinhibited eaters were characterised by a reduced HRV which was related to greater BG excursions (B = 0.407, 95% CI LL 0.044, UL 1.134). These findings highlight novel mechanisms by which disinhibited eating leads to obesity and insulin resistance. This trial was registered at clinicaltrials.gov NCT02827318.

(1) OBJECTIVE: To compare the effects of isomaltulose (Palatinose™, PSE) vs. maltodextrin (MDX) ingestion on substrate utilization during endurance exercise and subsequent time trial performance; (2) METHODS: 20 male athletes performed two experimental trials with ingestion of either 75 g PSE or MDX 45 min before the start of exercise. The exercise protocol consisted of 90 min cycling (60% VO₂max) followed by a time trial; (3) RESULTS: Time trial finishing time (-2.7%, 90% CI: ±3.0%, 89% likely beneficial; p = 0.147) and power output during the final 5 min (+4.6%, 90% CI: ±4.0%, 93% likely beneficial; p = 0.053) were improved with PSE compared with MDX. The blood glucose profile differed between trials (p = 0.013) with PSE resulting in lower glycemia during rest (95%-99% likelihood) and higher blood glucose concentrations during exercise (63%-86% likelihood). In comparison to MDX, fat oxidation was higher (88%-99% likelihood; p = 0.005) and carbohydrate oxidation was lower following PSE intake (85%-96% likelihood; p = 0.002). (4) CONCLUSION: PSE maintained a more stable blood glucose profile and higher fat oxidation during exercise which resulted in improved cycling performance compared with MDX. These results could be explained by the slower availability and the low-glycemic properties of Palatinose™ allowing a greater reliance on fat oxidation and sparing of glycogen during the initial endurance exercise.

Isomerization of disaccharides (maltose, isomaltose, cellobiose, lactose, melibiose, palatinose, sucrose, and trehalose) was investigated in subcritical aqueous ethanol. A marked increase in the isomerization of aldo-disaccharides to keto-disaccharides was noted and their hydrolytic reactions were suppressed with increasing ethanol concentration. Under any study condition, the maximum yield of keto-disaccharides produced from aldo-disaccharides linked by β-glycosidic bond was higher than that produced from aldo-disaccharides linked by α-glycosidic bond. Palatinose, a keto-disaccharide, mainly underwent decomposition rather than isomerization in subcritical water and subcritical aqueous ethanol. No isomerization was noted for the non-reducing disaccharides trehalose and sucrose. The rate constant of maltose to maltulose isomerization almost doubled by changing solvent from subcritical water to 80 wt% aqueous ethanol at 220 °C. Increased maltose monohydrate concentration in feed decreased the conversion of maltose and the maximum yield of maltulose, but increased the productivity of maltulose. The maximum productivity of maltulose was ca. 41 g/(h kg-solution).

Unlabelled: Effects of the dietary glycaemic load on postprandial blood glucose and insulin response might be of importance for fat deposition and risk of obesity. We aimed to investigate the metabolic effects, acceptance and tolerance of a follow-on formula containing the low glycaemic and low insulinaemic carbohydrate isomaltulose replacing high glycaemic maltodextrin. Healthy term infants aged 4 to 8 completed months (n = 50) were randomized to receive the intervention follow-on formula (IF, 2.1g isomaltulose (Palatinose™)/100mL) or an isocaloric conventional formula (CF) providing 2.1g maltodextrin/100mL for four weeks. Plasma insulinaemia 60 min after start of feeding (primary outcome) was not statistically different, while glycaemia adjusted for age and time for drinking/volume of meal 60 min after start of feeding was 122(105,140) mg/dL in IF (median, interquartile range) and 111(100,123) in CF (p = 0.01). Urinary c-peptide:creatinine ratio did not differ (IF:81.5(44.7, 96.0) vs. CF:56.8(37.5, 129),p = 0.43). Urinary c-peptide:creatinine ratio was correlated total intake of energy (R = 0.31,p = 0.045), protein (R = 0.42,p = 0.006) and fat (R = 0.40,p = 0.01) but not with carbohydrate intake (R = 0.22,p = 0.16). Both formulae were well accepted without differences in time of crying, flatulence, stool characteristics and the occurrence of adverse events. The expected lower postprandial plasma insulin and blood glucose level due to replacement of high glycaemic maltodextrin by low glycaemic isomaltulose were not observed in the single time-point blood analysis. In infants aged 4 to 8 completed months fed a liquid formula, peak blood glucose might be reached earlier than 60 min after start of feeding. Non-invasive urinary c-peptide measurements may be a suitable marker of nutritional intake during the previous four days in infants., Trial Registration: ClinicalTrials.gov NCT01627015.

Objective: To investigate the effect of isomaltulose (ISO) and sucrose (SUC) based breakfast on the postprandial substrate oxidation and glycemic/insulinemic changes in type 2 diabetes mellitus subjects (T2DM)., Material and Method: This was a randomized, controlled, double blind, and crossover study performed in two to five days. About 10 to 12 hours prior to the test, the subjects were not allowed to consume food or drink other than water. On the experimental day pre- and postprandial plasma glucose, serum insulin, and substrate utilization were measured after the subjects consumed a test breakfast with ISO or SUC, followed by a standard lunch three hours later., Results: The plasma glucose levels in subjects after consuming ISO breakfast tended to be lower than subjects consuming SUC breakfast at 30- and 60-minute, respectively. The second meal effect after standard lunch (240-minute) on insulin levels in subject consuming ISO breakfast tended to be lower than that of subjects consuming SUC breakfast. Substrate oxidation indicated that the incremental area under the fat oxidation of ISO breakfast was 20% higher comparing to SUC breakfast., Conclusion: Therefore, ISO based breakfast tends to provide less postprandial glucose and insulin levels than SUC based breakfast, thereby increasing postprandial fat oxidation.

The sucrose isomerase of Serratia plymuthica AS9 (AS9 PalI) was expressed in Escherichia coli BL21(DE3) and characterized. The half-life of AS9 PalI was 20 min at 45°C, indicating that it was unstable. In order to improve its thermostability, six amino acid residues with higher B-factors were selected as targets for site-directed mutagenesis, and six mutants (E175N, K576D, K174D, G176D, S575D and N577K) were designed using the RosettaDesign server. The E175N and K576D mutants exhibited improved thermostability in preliminary experiments, so the double mutant E175N/K576D was constructed. These three mutants (E175N, K576D, E175N/K576D) were characterized in detail. The results indicate that the three mutants exhibit a slightly increased optimal temperature (35°C), compared with that of the wild-type enzyme (30°C). The mutants also share an identical pH optimum of 6.0, which is similar to that of the wild-type enzyme. The half-lives of the E175N, K576D and E175N/K576D mutants were 2.30, 1.78 and 7.65 times greater than that of the wild-type enzyme at 45°C, respectively. Kinetic studies showed that the Km values for the E175N, K576D and E175N/K576D mutants decreased by 6.6%, 2.0% and 11.0%, respectively, and their kcat/Km values increased by 38.2%, 4.2% and 19.4%, respectively, compared with those of the wild-type enzyme. After optimizing the conditions for isomaltulose production at 45°C, we found that the E175N, K576D and E175N/K576D mutants displayed slightly improved isomaltulose yields, compared with the wild-type enzyme. Therefore, the mutants produced in this study would be more suitable for industrial biosynthesis of isomaltulose.

Certain commercial carbohydrate replacement products include slowly absorbed carbohydrates such as isomaltulose. Few studies have investigated the metabolic effects of ingesting isomaltulose during exercise and none have evaluated exercise performance and gastrointestinal comfort. Nine male cyclists participated postprandially during three trials of 2-h steady-state (S-S) exercise (60%Wmax) followed by a 16 km time trial (TT) while ingesting 63 g·h-1 of either, 0.8:1 fructose: maltodextrin (F:M) or isomaltulose (ISO) or placebo- flavored water (PL). Data were analyzed by magnitude-based inferences. During S-S exercise, ISO and PL similarly increased plasma nonesterified fatty acid (NEFA) concentration (mean change ISO versus F:M: 0.18, 90%CI ±0.21 mmol·L-1, 88% likelihood) and fat oxidation (10, 90%CI ±9 g, 89% likelihood) while decreasing carbohydrate oxidation (-36, 90%CI ±30.2 g, 91% likelihood) compared with F:M, despite equal elevations in blood glucose concentration with ISO and F:M. Rating of stomach cramps and bloating increased progressively with ISO (rating: 0-90 min S-S, weak; 120 min S-S, moderate; TT, strong) compared with F:M and PL (0-120 min S-S and TT, very weak). TT performance was substantially slower with ISO (mean change: 1.5, 90%CI ±1.4 min, 94% likely harmful) compared with F:M. The metabolic response of ISO ingestion during moderate exercise to increase NEFA availability and fat oxidation despite elevating blood glucose concentration is anomalous for a carbohydrate supplement. However, ingesting isomaltulose at a continuous high frequency to meet the recommended carbohydrate replacement dose, results in severe gastrointestinal symptoms during prolonged or high intensity exercise and negatively affects exercise performance compared with fructose-maltodextrin supplementation.

Purpose: Although previous research has associated the glycaemic load (GL) of a meal with cognitive functioning, typically the macro-nutrient composition of the meals has differed, raising a question as to whether the response was to GL or to the energy, nutrients or particular foods consumed. Therefore, the present study contrasted two breakfasts that offered identical levels of energy and macro-nutrients, although they differed in GL., Methods: Using a repeated-measures, double-blind design, 75 children aged 5-11 years, from socially deprived backgrounds, attended a school breakfast club and on two occasions, at least a week apart, they consumed a meal sweetened with either isomaltulose (Palatinose™) (GL 31.6) or glucose (GL 59.8). Immediate and delayed verbal memory, spatial memory, sustained attention, reaction times, speed of information processing and mood were assessed 1 and 3 h after eating., Results: The nature of the meals did not influence any measure of cognition or mood after an hour; however, after 3 h, children's memory and mood improved after the lower-GL breakfast. If children had eaten the lower-GL meal on the second day of testing, they were able to process information faster and had better spatial memory later in the morning., Conclusions: Towards the end of a morning in school, having consumed a lower-GL breakfast resulted in better mood and aspects of cognitive functioning.

Isomaltulose, also known as palatinose, is produced by sucrose isomerase and has been highlighted as a sugar substitute due to a number of advantageous properties. For the massive production of isomaltulose, high resistance to sucrose and stability of sucrose isomerase as well as sucrose conversion yields would be critical factors. We describe a series of screening procedures to isolate the mutant strain of Serratia sp. possessing enhanced isomaltulose production with improved stability. The new Serratia sp. isolated from a series of screening procedures allowed us to produce isomaltulose from 60% sucrose solution, with over 90% conversion yield. Moreover, when this strain was immobilized in calcium alginate beads and placed in a medium containing 60% sucrose, it showed over 70% sucrose conversion yields for 30 cycles of repeated-batch reactions. Thus, improved conversion activity and stability of the newly isolated Serratia sp. strain in the present study would be highly valuable for industries related to isomaltulose production.

Aims/hypothesis: High intake of carbohydrates, particularly sucrose, in western societies is associated with the development of non-alcoholic fatty liver (NAFL) and diabetes mellitus. It is unclear whether this is related primarily to the carbohydrate quantity or to the hormonal responses, particularly glucose-dependent insulinotropic polypeptide (GIP), which is released in the proximal intestine. Therefore, we investigated the role of GIP by comparing two glucose-fructose dimers, sucrose and Palatinose (isomaltulose), resorbed proximally or distally., Methods: The glycaemic and incretin responses to sucrose and Palatinose were studied by oral gavage and meal tests. We then analysed phenotypic and metabolic diet-induced changes in C57Bl/6J mice exposed to isoenergetic diets differing in carbohydrate type. Studies were repeated in GIP receptor knockout (Gipr(-/-)) mice and their wild-type littermates., Results: Compared with sucrose, Palatinose intake resulted in slower glucose absorption and reduced postprandial insulin and GIP levels. After 22 weeks, Palatinose feeding prevented hepatic steatosis (48.5%) compared with sucrose and improved glucose tolerance, without differences in body composition and food intake. Ablation of GIP signalling in Gipr(-/-) mice completely prevented the deleterious metabolic effects of sucrose feeding. Furthermore, our microarray analysis indicated that sucrose increased 2.3-fold the hepatic expression of Socs2, which is involved in the growth hormone signalling pathway and participates in the development of NAFL., Conclusions/interpretation: Our results suggest that the site of glucose absorption and the GIP response determine liver fat accumulation and insulin resistance. GIP may play a role in sucrose induced fatty liver by regulating the expression of Socs2.

Objective: This study evaluated whether a combination of whey protein (WP), calcium beta-hydroxy-beta-methylbutyrate (HMB), and carbohydrate exert additive effects on recovery from highly demanding resistance exercise., Methods: Thirteen resistance-trained men (age: 22.6 ± 3.9 years; height: 175.3 ± 12.2 cm; weight: 86.2 ± 9.8 kg) completed a double-blinded, counterbalanced, within-group study. Subjects ingested EAS Recovery Protein (RP; EAS Sports Nutrition/Abbott Laboratories, Columbus, OH) or WP twice daily for 2 weeks prior to, during, and for 2 days following 3 consecutive days of intense resistance exercise. The workout sequence included heavy resistance exercise (day 1) and metabolic resistance exercise (days 2 and 3). The subjects performed no physical activity during day 4 (+24 hours) and day 5 (+48 hours), where recovery testing was performed. Before, during, and following the 3 workouts, treatment outcomes were evaluated using blood-based muscle damage markers and hormones, perceptual measures of muscle soreness, and countermovement jump performance., Results: Creatine kinase was lower for the RP treatment on day 2 (RP: 166.9 ± 56.4 vs WP: 307.1 ± 125.2 IU · L(-1), p ≤ 0.05), day 4 (RP: 232.5 ± 67.4 vs WP: 432.6 ± 223.3 IU · L(-1), p ≤ 0.05), and day 5 (RP: 176.1 ± 38.7 vs 264.5 ± 120.9 IU · L(-1), p ≤ 0.05). Interleukin-6 was lower for the RP treatment on day 4 (RP: 1.2 ± 0.2 vs WP: 1.6 ± 0.6 pg · ml(-1), p ≤ 0.05) and day 5 (RP: 1.1 ± 0.2 vs WP: 1.6 ± 0.4 pg · ml(-1), p ≤ 0.05). Muscle soreness was lower for RP treatment on day 4 (RP: 2.0 ± 0.7 vs WP: 2.8 ± 1.1 cm, p ≤ 0.05). Vertical jump power was higher for the RP treatment on day 4 (RP: 5983.2 ± 624 vs WP 5303.9 ± 641.7 W, p ≤ 0.05) and day 5 (RP: 5792.5 ± 595.4 vs WP: 5200.4 ± 501 W, p ≤ 0.05)., Conclusions: Our findings suggest that during times of intense conditioning, the recovery benefits of WP are enhanced with the addition of HMB and a slow-release carbohydrate. We observed reductions in markers of muscle damage and improved athletic performance.

Background: Isomaltulose attenuates postprandial glucose and insulin concentrations compared with sucrose in patients with type 2 diabetes mellitus (T2DM). However, the mechanism by which isomaltulose limits postprandial hyperglycemia has not been clarified., Objective: The objective was therefore to assess the effects of bolus administration of isomaltulose on glucose metabolism compared with sucrose in T2DM., Design: In a randomized, double-blind, crossover design, 11 participants with T2DM initially underwent a 3-h euglycemic-hyperinsulinemic (0.8 mU · kg(-1) · min(-1)) clamp that was subsequently combined with 1 g/kg body wt of an oral (13)C-enriched isomaltulose or sucrose load. Hormonal responses and glucose kinetics were analyzed during a 4-h postprandial period., Results: Compared with sucrose, absorption of isomaltulose was prolonged by ∼50 min (P = 0.004). Mean plasma concentrations of insulin, C-peptide, glucagon, and glucose-dependent insulinotropic peptide were ∼10-23% lower (P < 0.05). In contrast, glucagon-like peptide 1 (GLP-1) was ∼64% higher (P < 0.001) after isomaltulose ingestion, which results in an increased insulin-to-glucagon ratio (P < 0.001) compared with sucrose. The cumulative amount of systemic glucose appearance was ∼35% lower after isomaltulose than after sucrose (P = 0.003) because of the reduction in orally derived and endogenously produced glucose and a higher first-pass splanchnic glucose uptake (SGU). Insulin action was enhanced after isomaltulose compared with sucrose (P = 0.013)., Conclusions: Ingestion of slowly absorbed isomaltulose attenuates postprandial hyperglycemia by reducing oral glucose appearance, inhibiting endogenous glucose production (EGP), and increasing SGU compared with ingestion of rapidly absorbed sucrose in patients with T2DM. In addition, GLP-1 secretion contributes to a beneficial shift in the insulin-to-glucagon ratio, suppression of EGP, and enhancement of SGU after isomaltulose consumption. This trial was registered at clinicaltrials.gov as NCT01070238., (© 2014 American Society for Nutrition.)

This paper reports the efficient enzymatic synthesis of a homologous series of isomaltulose-derived oligosaccharides with degrees of polymerization ranging from 3 to 9 through the transglucosylation reaction using a dextransucrase from Leuconostoc mesenteroides B-512F. The total oligosaccharide yield obtained under optimal conditions was 41-42% (in weight with respect to the initial amount of isomaltulose) after 24-48 h of reaction. Nuclear magnetic resonance (NMR) structural characterization indicated that dextransucrase specifically transferred glucose moieties of sucrose to the C-6 of the nonreducing glucose residue of isomaltulose. Likewise, monitoring the progression of the content of each individual oligosaccharide indicated that oligosaccharide acceptor products of low molecular weight acted in turn as acceptors for further transglucosylation to yield oligosaccharides of a higher degree of polymerization. The produced isomaltulose-derived oligosaccharides can be considered as isomalto-oligosaccharides (IMOs) because they are linked by only α-(1→6) bonds. In addition, having isomaltulose as the core structure, these IMO-like structures could possess appealing bioactive properties that could find potential applications as functional food ingredients.

Isomaltulose is a natural isomer of sucrose. It is widely used as a functional sweetener with promising properties, including slower digestion, lower glycemic index, prolonged energy release, lower insulin reaction, and less cariogenicity. It has been approved as a safe sucrose substitute by the Food and Drug Administration of the US; Ministry of Health, Labor and Welfare of Japan; and the Commission of the European Communities. This article presents a review of recent studies on the properties, physiological effects, and food application of isomaltulose. In addition, the biochemical properties of sucrose isomerases producing isomaltulose are compared; the heterologous expression, fermentation optimization, structural determination, and catalysis mechanism of sucrose isomerase are reviewed; and the biotechnological production of isomaltulose from sucrose is summarized.

We wished to clarify the inaccuracy of AOAC method 2009.01 for the measurement of non-digestible oligosaccharides and to propose an improved method using porcine intestinal enzymes. Amyloglucosidase used in AOAC method 2009.01 scarcely hydrolyses sucrose, palatinose and panose (which are readily digested by intestinal enzymes). Hence, oligosaccharides could not be measured accurately by AOAC method 2009.01. To confirm the inaccuracy of the method, we used porcine intestinal enzymes instead of amyloglucosidase. Using the improved method, fructooligosaccharide and galactooligosaccharide were measured accurately as non-digestible oligosaccharides, but sucrose, palatinose, panose and isomaltooligosaccharide were not. The improved method hydrolysed digestible oligosaccharides into monosaccharides. These results demonstrate that the inaccuracy of AOAC method 2009.01 for oligosaccharide measurement is due to incomplete hydrolysis by amyloglucosidase. We propose that amyloglucosidase should be replaced with porcine intestinal enzymes for such measurements., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Fifty-three plant-associated microorganisms were investigated for their ability to convert sucrose to its isomers. These microorganisms included one Dickeya zeae isolate and 7 Enterobacter, 3 Pantoea, and 43 Pectobacterium species. Eleven out of the 53 strains (21%) showed the ability to transform sucrose to isomaltulose and trehalulose. Among those, Pectobacterium carotovorum KKH 3-1 showed the highest bioconversion yield (97.4%) from sucrose to its isomers. In this strain, the addition of up to 14% sucrose in the medium enhanced sucrose isomerase (SIase) production. The SIase activity at 14% sucrose (47.6 U/mg dcw) was about 3.6-fold higher than that of the negative control (13.3 U/mg dcw at 0% sucrose). The gene encoding SIase, which is comprised a 1776 bp open reading frame (ORF) encoding 591 amino acids, was cloned from P. carotovorum KKH 3-1 and expressed in Escherichia coli. The recombinant SIase (PCSI) was shown to have optimum activity at pH 6.0 and 40 °C. The reaction temperature significantly affected the ratio of sucrose isomers produced by PCSI. The amount of trehalulose increased from 47.5% to 79.1% as temperature was lowered from 50 °C to 30 °C, implying that SIase activity can be controlled by reaction temperature., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Success in creating a synbiotic depends on compatibility between the chosen components--prebiotic and probiotic. In this work the interactions between Lactobacillus sp. strains isolated from yogurts and type strains of Lactobacillus sp. and Lactococcus sp., and the dependence of their growth and antibacterial activity on three oligosaccharides (OS)--palatinose, inulin and alpha-cyclodextrin were investigated. All isolated lactobacilli produce antibacterial compounds, which possibly are the bacteriocins of Lactobacillus casei ATCC334 strain. Results of growth analysis with different OS revealed that part of lactobacilli isolated from yogurts can effectively ferment inulin and may be used for the development of synbiotics. Palatinose and Lactobacillus acidophilus could be used as symbiotics with effective antibacterial activity. One of the types of Lactococcus sp. strains can assimilate palatinose and alpha-cyclodextrin, so they both can be used as components of synbiotics with the investigated lactococci. Results of this analysis suggest that the investigated isolated and type strains of Lactobacillus sp. and Lactoccocus sp. can be useful as probiotics in the development of synbiotics. Together with prebiotics--palatinose, inulin and alpha-cyclodextrin, the synbiotics, which could regulate not only the growth of beneficial bacteria in the gastrointestinal tract, but also their antibacterial activity, can be created.

The consumption of carbohydrate before, during, and after exercise is a central feature of the athlete's diet, particularly those competing in endurance sports. Sucrose is a carbohydrate present within the diets of athletes. Whether sucrose, by virtue of its component monosaccharides glucose and fructose, exerts a meaningful advantage for athletes over other carbohydrate types or blends is unclear. This narrative reviews the literature on the influence of sucrose, relative to other carbohydrate types, on exercise performance or the metabolic factors that may underpin exercise performance. Inference from the research to date suggests that sucrose appears to be as effective as other highly metabolizable carbohydrates (e.g., glucose, glucose polymers) in providing an exogenous fuel source during endurance exercise, stimulating the synthesis of liver and muscle glycogen during exercise recovery and improving endurance exercise performance. Nonetheless, gaps exist in our understanding of the metabolic and performance consequences of sucrose ingestion before, during, and after exercise relative to other carbohydrate types or blends, particularly when more aggressive carbohydrate intake strategies are adopted. While further research is recommended and discussed in this review, based on the currently available scientific literature it would seem that sucrose should continue to be regarded as one of a variety of options available to help athletes achieve their specific carbohydrate-intake goals.

Glycaemic response to dietary carbohydrates might have an impact on cognitive performance. The present study investigated the effects of growing-up milks (GUM) with isomaltulose and extra minerals and vitamins or lower protein content on cognitive parameters in children aged 5–6 years. In a blinded, partly randomised, controlled, cross-over study, four GUM were provided, each taken over 14 d (2 × 200 ml/d): standard (Std) GUM; Std GUM+5 g isomaltulose (Iso-5 GUM); Iso-5 GUM with 26 % less protein (Iso-5 LP GUM); Std GUM with 2·5 g isomaltulose and extra Mg, Zn, Se, D3, B1, B2, B12, folic acid and choline (Iso-2·5 GUM). At test days, when GUM replaced breakfast, repeated (0, 60, 120 and 180 min post-dose) cognitive tasks were performed (picture presentation, simple reaction time, digit vigilance, choice reaction time, spatial and numeric working memory and picture recognition). Task performance of all subjects (n 50) worsened over the morning. Best performance was seen on isomaltulose GUM, most notably at 180 min. Iso-2·5 GUM showed best performance on several parameters of attention and memory, Iso-5 GUM performed best on parameters of memory and Iso-5 LP GUM was positively associated with parameters of attention but less with memory. Std GUM showed only a benefit on one attention and one memory task. Thus, isomaltulose-enriched GUM positively affected parameters of attention and memory at 180 min post-dose when compared with Std GUM. Extra minerals and vitamins seem beneficial, whereas lowering protein content might improve attention in particular.

A new strawberry spread formulated with fructose and isomaltulose (replacing sucrose partially or totally) and a high percentage of fruit was developed in line with the new trend of healthier products. This work studies the influence of some process variables (percentage of sugar, pectin and citric acid, and time of thermal treatment) on the volatile profile of these spreads with different formulations. The ripeness of the raw strawberries influences the concentrations of some of the compounds in the spreads, such as isobutyl acetate, butyl butyrate, 3-hexen-1-yl acetate or propan-2-ol. The process conditions have an important effect on the volatile profiles. Most of the esters and alcohols decreased whereas 13 new compounds appear, mostly furans (furfural, 2-acetylfurane, 5-methyl furfural, mesifurane) and aldehydes (octanal, nonanal, decanal and benzaldeyhde). In general, the spreads formulated with sucrose-isomaltulose that contained higher levels of pectin and citric acid gave better results in the preservation of the original aromatic compounds in raw strawberries., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Isomaltulose (IM) is a natural isomer of sucrose. It is widely approved as a food with properties including slower digestion, lower glycaemic index and low cariogenicity, which can benefit consumers. Availability is currently limited by the cost of fermentative conversion from sucrose. Transgenic sugarcane plants with developmentally-controlled expression of a silencing-resistant gene encoding a vacuole-targeted IM synthase were tested under field conditions typical of commercial sugarcane cultivation. High yields of IM were obtained, up to 483 mm or 81% of total sugars in whole-cane juice from plants aged 13 months. Using promoters from sugarcane to drive expression preferentially in the sugarcane stem, IM levels were consistent between stalks and stools within a transgenic line and across consecutive vegetative field generations of tested high-isomer lines. Germination and early growth of plants from setts were unaffected by IM accumulation, up to the tested level around 500 mm in flanking stem internodes. These are the highest yields ever achieved of value-added materials through plant metabolic engineering. The sugarcane stem promoters are promising for strategies to achieve even higher IM levels and for other applications in sugarcane molecular improvement. Silencing-resistant transgenes are critical to deliver the potential of these promoters in practical sugarcane improvement. At the IM levels now achieved in field-grown sugarcane, direct production of IM in plants is feasible at a cost approaching that of sucrose, which should make the benefits of IM affordable on a much wider scale., (© 2013 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.)

Sucrose isomerase is an enzyme that catalyzes the production of sucrose isomers of high biotechnological and pharmaceutical interest. Owing to the complexity of the chemical synthesis of these isomers, isomaltulose and trehalulose, enzymatic conversion remains the preferred method for obtaining these products. Depending on the microbial source, the ratio of the sucrose-isomer products varies significantly. In studies aimed at understanding and explaining the underlying molecular mechanisms of these reactions, mutations obtained using a random-mutagenesis approach displayed a major hydrolytic activity. Two of these variants, R284C and F164L, of sucrose isomerase from Rhizobium sp. were therefore crystallized and their crystal structures were determined. The three-dimensional structures of these mutants allowed the identification of the molecular determinants that favour hydrolytic activity compared with transferase activity. Substantial conformational changes resulting in an active-site opening were observed, as were changes in the pattern of water molecules bordering the active-site region.

Objective: A combined load of carbohydrate and protein stimulates insulin secretion. However, results on postprandial glucose responses in type 2 diabetic (T2D) subjects have been inconclusive. Therefore, we investigated the effects of co-ingestion of carbohydrate and protein on glucose and insulin responses in these subjects., Methods: After an overnight fast, 30 subjects consumed a drink containing 50 g of slowly-digested isomaltulose (ISO), combined either with a mixture of 21 g whey/soy (ISO+WS) or with 21 g casein (ISO+C) in a randomized order on separate days. In another experiment, the subjects consumed a control drink containing only 50 g ISO., Results: No significant differences in glucose responses were observed after ingestion of the drinks. Compared to ingestion of ISO alone, insulin response was ~190%-270% higher (P<.001), whereas insulin action was lower (P<.01) after ingestion of ISO+WS and ISO+C. Plasma insulin levels increased more significantly (P<.001) after ingestion of ISO+WS compared to ISO+C and were positively correlated with total amino acid levels (P<.001). Insulin action, however, showed a greater decrease following ingestion of ISO+WS than ISO+C (P<.01)., Conclusions: Combining carbohydrate with protein can elevate postprandial insulin levels, but decreases insulin action, and therefore does not improve glucose response in T2D subjects. Our results further suggest that different types of proteins (i.e., fast-absorbing whey/soy vs. slow-absorbing casein) differently modulate insulin response and insulin action. A fast-absorbing protein mixture reduces insulin action to a greater extent than a slow-absorbing protein, and therefore may not be recommended for glycemic control in T2D patients., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The impact of slowly digestible sugars in reducing the risk of developing obesity and related metabolic disorders remains unclear. We hypothesised that such carbohydrates (CHO), resulting in a lower glycaemic and insulinaemic response, may lead to greater postprandial fat oxidation rates in subjects with impaired glucose tolerance (IGT). The present study intends to compare the postprandial metabolic responses to the ingestion of glucose (GLUC) v. trehalose (TRE) and sucrose (SUC) v. isomaltulose (IMU). In a randomised, single-blind, cross-over design, ten overweight IGT subjects were studied four times, following ingestion of different CHO drinks either at breakfast or in combination with a mixed meal at lunch. Before and 3 h after CHO ingestion, energy expenditure, substrate utilisation and circulating metabolite concentrations were determined. Ingestion of CHO drinks with a meal resulted in an attenuated rise in GLUC (-33 %) and insulin (-14 %) concentrations following TRE when compared with GLUC and following IMU, an attenuation of 43 and 34 % when compared with SUC ingestion, respectively. Additionally, there was less inhibition of the rise in NEFA concentrations and less decline in postprandial fat oxidation (22 %) after IMU when compared with SUC, whereas TRE did not differ from GLUC. The attenuated rise in GLUC and insulin concentrations following IMU ingestion attenuated the postprandial inhibition of fat oxidation compared with SUC when co-ingested with a meal. This suggests that exchange of SUC in the diet for IMU may result in a more favourable metabolic response and may help to reduce the risks associated with obesity and type 2 diabetes.

Bud burst is a decisive process in plant architecture that requires light in Rosa sp. This light effect was correlated with stimulation of sugar transport and metabolism in favor of bud outgrowth. We investigated whether sugars could act as signaling entities in the light-mediated regulation of vacuolar invertases and bud burst. Full-length cDNAs encoding two vacuolar invertases (RhVI1 and RhVI2) were isolated from buds. Unlike RhVI2, RhVI1 was preferentially expressed in bursting buds, and was up-regulated in buds of beheaded plants exposed to light. To assess the importance of sugars in this process, the expression of RhVI1 and RhVI2 and the total vacuolar invertase activity were further characterized in buds cultured in vitro on 100 mM sucrose or mannitol under light or in darkness for 48 h. Unlike mannitol, sucrose promoted the stimulatory effect of light on both RhVI1 expression and vacuolar invertase activity. This up-regulation of RhVI1 was rapid (after 6 h incubation) and was induced by as little as 10 mM sucrose or fructose. No effect of glucose was found. Interestingly, both 30 mM palatinose (a non-metabolizable sucrose analog) and 5 mM psicose (a non-metabolizable fructose analog) promoted the light-induced expression of RhVI1 and total vacuolar invertase activity. Sucrose, fructose, palatinose and psicose all promoted bursting of in vitro cultured buds under light. These findings indicate that soluble sugars contribute to the light effect on bud burst and vacuolar invertases, and can function as signaling entities.

Objective: To test the hypothesis that replacement of sucrose with isomaltulose in sweet foods and beverages improves metabolic control in patients with type 2 diabetes., Research Design and Methods: One hundred ten patients with type 2 diabetes were randomized to receive sweet foods containing either 50 g/day isomaltulose or sucrose for 12 weeks as part of their habitual diet under free-living conditions. HbA(1c) at 12 weeks was the primary outcome parameter., Results: In the final analysis comprising 101 patients, isomaltulose did not significantly affect HbA(1c) at 12 weeks (sucrose: 7.39 ± 0.78%; isomaltulose: 7.24 ± 0.76%; regression coefficient [b]: 0.02 [95% CI: -0.21 to 0.25], P = 0.844). Triglycerides at 12 weeks were significantly lower in the isomaltulose versus the sucrose group (b: 34.01 [6.59-61.44], P = 0.016). Other secondary parameters did not significantly differ between groups., Conclusions: Isomaltulose did not influence glycemic control assessed as HbA(1c) in type 2 diabetes under free-living conditions but was associated with lower triglyceride levels.

Objective: Dietary interventions with a low glycemic index have shown to be successful for the prevention and therapy of the metabolic syndrome. In the present study, we investigated the postprandial metabolic response at rest and during physical activity the low glycemic carbohydrate isomaltulose (Palatinose™) intake compared with a conventional carbohydrate (glucose syrup/sucrose [glc/suc]) with a higher glycemic index., Methods: Twenty overweight or obese men (32-64 y old) with the metabolic syndrome and insulin resistance were enrolled in this double-blinded, randomized, cross-over study. In the morning, a breakfast consisting of a 250-mL drink and 140 g of cookies containing in a total of 50 g of Palatinose™ or glc/suc was consumed. Two hours after breakfast, subjects exercised at moderate intensity on a treadmill for 30 min. Thereafter, subjects ingested a standardized lunch consisting of a 250-mL drink with 10% Palatinose™ or glc/suc, mini pizzas, and an apple., Results: Blood levels of glucose and insulin were measured and the postprandial substrate metabolism was determined. The glycemic and insulinemic responses were considerably lower after the ingestion of Palatinose™ (incremental area under the curve, P < 0.05). The total fat oxidation was significantly higher with Palatinose™ from breakfast to the beginning of lunch including the exercise and postexercise periods (P < 0.05). Fat oxidation with Palatinose™ was numerically higher throughout the entire examination period (P = 0.09)., Conclusion: In obese subjects with insulin resistance and the metabolic syndrome, the partial substitution of carbohydrates with a higher glycemic index in foods and drinks by Palatinose™ resulted in greater postprandial fat oxidation at rest and during physical activity. It is hypothesized that this increased fat oxidation may confer further benefits for long-term weight management and for an improvement in metabolic risk factors., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Purpose: Exercise improves insulin resistance and is a first line for the prevention and treatment of type 2 diabetes. The extent, however, to which these responses are dose dependent is not known. The purpose of this study was to examine whether exercise dose was associated with improvements in insulin sensitivity after 4 months of exercise training in previously sedentary adults., Methods: Fifty-five healthy volunteers participated in a 16-wk supervised endurance exercise intervention with a pre/postintervention design. Insulin sensitivity was assessed by euglycemic hyperinsulinemic clamp, peak oxygen uptake by a graded exercise test, and body composition by dual-energy x-ray absorptiometry. The exercise intervention consisted of three to five sessions per week with a minimum of three sessions supervised. A ramped exercise prescription protocol was used to achieve 75% of peak HR for 45 min per session. Exercise dose, expressed as average kilocalories expended per week, was computed as the product of exercise intensity, duration and frequency., Results: Improved insulin sensitivity was significantly related to exercise dose in a graded dose-response relationship. No evidence of threshold or maximal dose-response effect was observed. Age and gender did not influence this dose-response relationship. Exercise intensity was also significantly related to improvements in insulin sensitivity, whereas frequency was not., Conclusions: This study identifies a graded dose-response relationship between exercise dose and improvements in insulin sensitivity. The implication of this observation is of importance for the adaptation of exercise prescription in clinical situations.

Purpose: Individuals with type 1 diabetes mellitus (T1DM) are encouraged to reduce rapid-acting insulin and consume CHO to prevent hypoglycemia during or after exercise. However, research comparing the metabolic and performance effects of different CHO is limited. This study compared the alterations in metabolism and fuel oxidation in response to performance running after preexercise ingestion of isomaltulose or dextrose in T1DM., Methods: After preliminary testing, on two occasions, seven T1DM individuals consumed 0.6 g·kg⁻¹ body mass of either dextrose (DEX; glycemic index = 96) or isomaltulose (ISO; glycemic index = 32), 2 h before a discontinuous incremental run to 80% V˙O2peak on a motorized treadmill followed by a 10-min all-out performance test on a nonmotorized treadmill. Blood glucose (BG), acid-base, and cardiorespiratory parameters were measured 2 h before, during, and after both run tests. Data (mean ± SEM) were analyzed using repeated-measures ANOVA., Results: Preexercise BG area under the curve was lower under ISO in comparison with DEX (ISO = +4.0 ± 0.3 mmol·L⁻¹·h⁻¹ vs DEX = +7.0 ± 0.6 mmol·L⁻¹·h⁻¹, P < 0.01). Resting blood lactate concentrations and rate of CHO oxidation under ISO were greater than those elicited under DEX (P < 0.05). There were no metabolic or cardiorespiratory differences between conditions in response to submaximal exercise despite lower BG concentrations under ISO (P < 0.05). T1DM individuals completed the same distance at the same speed during the 10-min run test under both conditions (not significant)., Conclusions: Consumption of isomaltulose alongside rapid-acting insulin reduction improves BG responses to exercise and produces a similar high-intensity run performance compared with dextrose in T1DM individuals.

Two different techniques of glucosyltransferase immobilization were studied for the conversion of sucrose into isomaltulose. The optimum conditions for immobilization of Erwinia sp. glucosyltransferase onto Celite 545, determined using response surface methodology, was pH 4.0 and 170 U of glucosyltransferase/g of Celite 545. Using this conditions more than 60% conversion of sucrose into isomaltulose can be obtained. The immobilization of glucosyltransferase was also studied by its entrapment in microcapsules of low-methoxyl pectin and fat (butter and oleic acid). The non-lyophilized microcapsules of pectin, containing the enzyme and fat, showed higher glucosyltransferase activity, compared with lyophilized microcapsules containing enzyme plus fat, and also lyophilized microcapsules containing enzyme without fat addition. The non-lyophilized microcapsules of pectin containing the glucosyltransferase and fat, converted 30% of sucrose into isomaltulose in the first batch. However the conversion decreased to 5% at the 10th batch, indicating inactivation of the enzyme., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Transgenic sugarcane plants expressing a vacuole-targeted isomaltulose (IM) synthase in seven recipient genotypes (elite cultivars) were evaluated over 3 years at a field site typical of commercial cane growing conditions in the Burdekin district of Australia. IM concentration typically increased with internode maturity and comprised up to 217 mm (33% of total sugars) in whole-cane juice. There was generally a comparable decrease in sucrose concentration, with no overall decrease in total sugars. Sugarcane is vegetatively propagated from stem cuttings known as setts. Culture-derived plants were slower to establish and generally gave shorter and thinner stalks at harvest than those grown from field-sourced setts in the initial field generations. However, after several cycles of field propagation, selections were obtained with cane yields similar to the recipient genotypes. There was no apparent adverse effect of IM accumulation on vigour assessed by stalk height and diameter or other visual indicators including germination of setts and establishment of stools. There was some inconsistency in IM levels in juice, between samplings of the vegetatively propagated transgenic lines. Until the causes are resolved, it is prudent to selectively propagate from stalks with higher IM levels in the initial vegetative field generations. Pol/Brix ratio allowed rapid identification of lines with high IM levels, using common sugar industry instruments. Sucrose isomerase activity was low in these transgenic lines, and the results indicate strong potential to develop sugarcane for commercial-scale production of IM if higher activity can be engineered in appropriate developmental patterns., (© 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.)

Changes in blood glucose are hypothesized to influence cognitive performance and these changes can be affected by certain nutrients. This double-blind 4-period cross-over study evaluated the effects of a slow-release modified sucrose (isomaltulose) in combination with a high concentration of lactose on cognitive performance of 5-6 year old children. Thirty children received a standard growing upmilk (Std GUM), reformulated growing up milk (Reform GUM), standard growing up milk with lactose-isomaltulose (Iso GUM), and a standard glucose drink (Glucose). The CDR System, a computerised cognitive assessment system, was used to assess various measures of attention and memory of the children at baseline (T=0), 60 (T=1), 120 (T=2), and 180 (T=3) minutes following the intake of test products. Overall, there was a decline in performance over the morning on almost every cognitive task. Children showed better attention following consumption of Iso GUM compared to Std GUM but attention was not significantly different than Reform GUM and glucose. Also, Iso GUM conferred a beneficial effect over both Reform GUM and glucose on sensitivity index of numeric working memory with no difference observed between Iso GUM and Std GUM. Surprisingly, glucose group showed lowest decline in the sensitivity index of spatial working memory and highest speed in picture recognition, although the latter was significantly better than Reform GUM only. For speed of spatial working memory, Reform GUM had the lowest decline but was significantly different only with Std GUM. There was, however, no significant difference among conditions for continuity of attention, speed of numeric working memory and picture recognition sensitivity. Despite the small sample size, the findings are intriguing as carbohydrate composition seems to influence some aspects of cognitive performance such as attention and memory. However, further studies are needed to confirm these findings., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The human gastrointestinal tract can be positively modulated by dietary supplementation of probiotic bacteria in combination with prebiotic carbohydrates. Here differential transcriptomics and functional genomics were used to identify genes in Lactobacillus acidophilus NCFM involved in the uptake and catabolism of 11 potential prebiotic compounds consisting of α- and β-linked galactosides and glucosides. These oligosaccharides induced genes encoding phosphoenolpyruvate-dependent sugar phosphotransferase systems (PTS), galactoside pentose hexuronide (GPH) permease, and ATP-binding cassette (ABC) transporters. PTS systems were upregulated primarily by di- and tri-saccharides such as cellobiose, isomaltose, isomaltulose, panose and gentiobiose, while ABC transporters were upregulated by raffinose, Polydextrose, and stachyose. A single GPH transporter was induced by lactitol and galactooligosaccharides (GOS). The various transporters were associated with a number of glycoside hydrolases from families 1, 2, 4, 13, 32, 36, 42, and 65, involved in the catabolism of various α- and β-linked glucosides and galactosides. Further subfamily specialization was also observed for different PTS-associated GH1 6-phospho-β-glucosidases implicated in the catabolism of gentiobiose and cellobiose. These findings highlight the broad oligosaccharide metabolic repertoire of L. acidophilus NCFM and establish a platform for selection and screening of both probiotic bacteria and prebiotic compounds that may positively influence the gastrointestinal microbiota.

Isomaltulose is a structural isomer of sucrose (Suc). It has been widely used as a nonmetabolized sugar in physiological studies aimed at better understanding the regulatory roles and transport of sugars in plants. It is increasingly used as a nutritional human food, with some beneficial properties including low glycemic index and acariogenicity. Cloning of genes for Suc isomerases opened the way for direct commercial production in plants. The understanding that plants lack catabolic capabilities for isomaltulose indicated a possibility of enhanced yields relative to Suc. However, this understanding was based primarily on the treatment of intact cells with exogenous isomaltulose. Here, we show that sugarcane (Saccharum interspecific hybrids), like other tested plants, does not readily import or catabolize extracellular isomaltulose. However, among intracellular enzymes, cytosolic Suc synthase (in the breakage direction) and vacuolar soluble acid invertase (SAI) substantially catabolize isomaltulose. From kinetic studies, the specificity constant of SAI for isomaltulose is about 10% of that for Suc. Activity varied against other Suc isomers, with V(max) for leucrose about 6-fold that for Suc. SAI activities from other plant species varied substantially in substrate specificity against Suc and its isomers. Therefore, in physiological studies, the blanket notion of Suc isomers including isomaltulose as nonmetabolized sugars must be discarded. For example, lysis of a few cells may result in the substantial hydrolysis of exogenous isomaltulose, with profound downstream signal effects. In plant biotechnology, different V(max) and V(max)/K(m) ratios for Suc isomers may yet be exploited, in combination with appropriate developmental expression and compartmentation, for enhanced sugar yields.

The gene encoding sucrose isomerase from Enterobacter sp. FMB-1 species (ESI) was displayed on the cell surface of Saccharomyces cerevisiae EBY100 using a glycosylphosphatidylinositol (GPI) anchor attachment signal sequence. Fluorescence activated cell sorting (FACS) analysis and immunofluorescence microscopy confirmed the localization of ESI on the yeast cell surface. The displayed ESI (dESI) was stable at a broad range of temperatures (35-55 °C) and pHs (pH 5-7) with optimal temperature and pH at 45 °C and pH 7.0, respectively. In addition, the thermostability of the dESI was significantly enhanced compared with the recombinant ESI expressed in Escherichia coli. Biotransformation of sucrose to isomaltulose was observed in various ranges of substrate concentrations (50-250 mM) with a 6.4-7.4% conversion yield. It suggested that the bioconversion of sucrose to isomaltulose can be successfully performed by the dESI on the surface of host S. cerevisiae., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Background & Aims: Gut ischemia-reperfusion (I/R) is considered an important mechanism underlying multiple organ failure after severe surgical insults. We previously demonstrated an enteral diet enriched with whey-hydrolyzed peptide, fermented milk, and isomaltulose to have anti-inflammatory effects in a concanavalin A-induced hepatitis model. Here, we investigated whether the immune-modulating diet (IMD), could prevent systemic inflammation, thereby improving survival in a gut I/R model., Methods: Mice were randomized into control enteral diet (n = 58) or IMD (n = 56) for 2 weeks' feeding. In experiment 1, 39 mice underwent 45 min of gut ischemia, and were sacrificed at 3 h after reperfusion to collect blood samples. Plasma IL-6 and glucose levels were measured. In experiment 2, 75 mice underwent 60 min of ischemia, and their survival was observed until 48 h., Results: Plasma IL-6 and glucose levels of the IMD group were significantly lower than those of control mice. In association with these changes, the IMD improved survival rate at early time points (12 and 30 h) after gut I/R (p < 0.05, χ(2) test)., Conclusions: Nutritional management with the IMD may be useful for preventing systemic inflammatory response after gut I/R., (Copyright © 2011 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.)

Purpose: individuals with type 1 diabetes mellitus (T1DM) are encouraged to consume CHO to prevent hypoglycemia during or after exercise. However, the research comparing specific types of CHO is limited. This study compared the alterations in metabolism and fuel oxidation in response to running after preexercise ingestion of isomaltulose or dextrose in T1DM., Methods: after preliminary testing, on two occasions, eight T1DM individuals consumed 75 g of either dextrose (DEX; GI = 96) or isomaltulose (ISO; GI = 32), 2 h before performing 45 min of treadmill running at 80% ± 1% VO(2peak). Blood glucose (BG) was measured for 2 h before and 3 h after exercise. Cardiorespiratory parameters were collected at rest and during exercise. Data (mean ± SEM) were analyzed using repeated-measures ANOVA., Results: there was a smaller increase in BG in the preexercise period under ISO with peak BG occurring at 120 min after ingestion compared with 90 min under DEX (Δ+4.5 ± 0.4 vs Δ+9.1 ± 0.6 mmol·L, P < 0.01). During the final 10 min of exercise, there were lower CHO (ISO 2.85 ± 0.07 vs DEX 3.18 ± 0.08 g·min, P < 0.05) and greater lipid oxidation rates (ISO 0.33 ± 0.03 vs DEX 0.20 ± 0.03 g·min, P < 0.05) under ISO. After exercise, ISO BG was lower than DEX for the entire 180-min period, with BG area under the curve and mean BG concentrations being 21% ± 3% and 3.0 ± 0.4 mmol·L lower, respectively (P < 0.05)., Conclusions: consumption of ISO improves BG responses during and after exercise through reduced CHO and improved lipid oxidation during the later stages of exercise.