Difructose anhydride III: a 50-year perspective on its production and physiological functions.

Production and applications of difructose anhydride III (DFA-III) have attracted considerable attention because of its versatile physiological functions. Recently, large-scale production of DFA-III has been continuously explored, which opens a horizon for applications in the food and pharmaceutical industries. This review updates recent advances involving DFA-III, including: biosynthetic strategies, purification, and large-scale production of DFA-III; physiological functions of DFA-III and related mechanisms; DFA-III safety evaluations; present applications in food systems, existing problems, and further research prospects. Currently, enzymatic synthesis of DFA-III has been conducted both industrially and in academic research. Two biosynthetic strategies for DFA-III production are summarized: single- and double enzyme-mediated. DFA-III purification is achieved via yeast fermentation. Enzyme membrane bioreactors have been applied to meet the large-scale production demands for DFA-III. In addition, the primary physiological functions of DFA-III and their underlying mechanisms have been proposed. However, current applications of DFA-III are limited. Further research regarding DFA-III should focus on commercial production and purification, comprehensive study of physiological properties, extensive investigation of large-scale human experiments, and expansion of industrial applications. It is worthy to dig deep into potential application and commercial value of DFA-III.

Ingestion of difructose anhydride III partially suppresses the deconjugation and 7α-dehydroxylation of bile acids in rats fed with a cholic acid-supplemented diet.

Difructose anhydride III (DFAIII) is a prebiotic involved in the reduction of secondary bile acids (BAs). We investigated whether DFAIII modulates BA metabolism, including enterohepatic circulation, in the rats fed with a diet supplemented with cholic acid (CA), one of the 12α-hydroxylated BAs. After acclimation, the rats were fed with a control diet or a diet supplemented with DFAIII. After 2 weeks, each group was further divided into two groups and was fed diet with or without CA supplementation at 0.5 g/kg diet. BA levels were analyzed in aortic and portal plasma, liver, intestinal content, and feces. As a result, DFAIII ingestion reduced the fecal deoxycholic acid level via the partial suppression of deconjugation and 7α-dehydroxylation of BAs following CA supplementation. These results suggest that DFAIII suppresses production of deoxycholic acid in conditions of high concentrations of 12α-hydroxylated BAs in enterohepatic circulation, such as obesity or excess energy intake. Abbreviation: BA: bile acid; BSH: bile salt hydrolase; CA: cholic acid; DCA: deoxycholic acid; DFAIII: difructose anhydride III; MCA: muricholic acid; MS: mass spectrometry; NCDs: non-communicable diseases; LC: liquid chromatography; SCFA: short-chain fatty acid; TCA: taurocholic acid; TCDCA: taurochenodeoxycholic acid; TDCA: taurodeoxycholic acid; TUDCA: tauroursodeoxychlic acid; TαMCA: tauro-α-muricholic acid; TßMCA: tauro-ß-muricholic acid; TωMCA: tauro-ω-muricholic acid.

Effects of difructose anhydride III on serum immunoglobulin G concentration and health status of newborn Holstein calves during the preweaning period.

This experiment was performed to investigate the effects of increases in passively acquired immunoglobulin G (IgG) by difructose anhydride (DFA) III supplementation on subsequent serum IgG concentration and health status in calves during the preweaning period. Thirty newborn female Holstein calves were paired by birth order, and 2 calves in each pair were fed 2 L of the same batch of colostrum within 2 h and at 10 h after birth, and followed by 2 L of the same batch of pooled colostrum at 20 h after birth. One calf from each pair was assigned to the control (n = 15) or treatment (n = 15) group. All calves in the treatment group received 18 g of DFA III at each feeding from birth to 7 d of age, whereas calves in the control group did not receive DFA III. Blood samples were collected before feeding at 0, 10, 20, and 36 h, and 4 and 7 d of age, and sampling was repeated at 7-d intervals thereafter until 49 d of age for serum IgG analysis. Calves were monitored daily for diarrhea and respiratory diseases. Serum IgG concentrations peaked at 36 h of age in both groups. Apparent efficiency of IgG absorption and peak serum IgG concentration were higher in the treatment group than in the control group. Using multiple regression analysis, we showed that peak serum IgG concentration in the newborn calves was positively correlated with colostral IgG concentration and DFA III supplementation. Moreover, peak serum IgG concentration (36 h of age) positively influenced subsequent serum IgG concentration until 35 d of age for all calves in both groups. The treatment group had higher serum IgG concentration from 20 h to 21 d of age than the control group. However, we detected no differences between the groups in number of calves with diarrhea or respiratory disease.

Impact of difructose anhydride III, raffinose, and fructooligosaccharides on energy intake, gut hormones, and cecal fermentation in rats fed a high-fat and high-sucrose diet.

We investigated the effects of dietary supplementation of difructose anhydride III (DFA III), raffinose (Raf), and fructooligosaccharides (FOS) on diet-induced obesity development. Male rats were fed normal or high-fat and high-sucrose (HFS) diet, with or without supplementing (3%) DFA III, Raf, or FOS, for 8 or 5 weeks. Supplementing DFA III to the HFS diet decreased energy intake compared to the non-supplemented HFS diet. Accordingly, body weight gain and fat accumulation reduced in DFA III-fed rats. Cecal acetate production and plasma glucagon-like peptide-1 (GLP-1) and peptide-YY (PYY) were elevated in DFA III-fed rats, while Raf and FOS partially affected these parameters. These results demonstrate that DFA III has suppressive effect on excessive energy intake driven by the palatable obesogenic diet, possibly due to combined effects of increased anorexigenic factors such as cecal acetate production and GLP-1/PYY secretion.

Difructose anhydride III decreases body fat as a low-energy substitute or by decreasing energy intake in non-ovariectomized and ovariectomized female rats.

We evaluated the effects of difructose anhydride III (DFAIII) on body weights of ovariectomized rats, which are a good model for obesity by estrogen deficiency-induced overeating. Female rats (10 weeks old) were subjected to ovariectomy or sham operation and then fed with or without a diet containing 3% or 6% DFAIII for 33 days or pair-fed control diet during the same period. Rats fed DFAIII showed significantly decreased food intake, energy intake, body weight gain, body energy accumulation, and fat tissue weight than control group, regardless of ovariectomy. DFAIII may decrease body fat dependent of reduced food/energy intake. Compared with the respective pair feeding groups, rats fed DFAIII showed significantly decreased body energy and fat tissue weight, regardless of ovariectomy, suggesting its potential as a low-energy substitute for high-energy sweeteners. The low energy of DFAIII may contribute to decreased body fat, which may not be dependent on obesity.

Effect of difructose anhydride III supplementation on passive immunoglobulin G transfer and serum immunoglobulin G concentration in newborn Holstein calves fed pooled colostrum.

Difructose anhydride (DFA) III acts on intestinal epithelial tight junctions and promotes calcium absorption through the paracellular pathway in dairy cows. This trial was done to investigate the effects of DFA III supplementation on passive IgG transfer and serum IgG concentration in calves. Twenty-four newborn Holstein male calves were grouped as 12 pairs, with each pair receiving a separate batch of pooled colostrum (PC) from the first 2 milkings. One calf from each pair was allocated to the treatment group (n=12) and received 18g of DFA III in each feeding; the other calf was assigned to the control group (n=12) and did not receive DFA III. All calves were fed 2 L of PC per feeding at 1, 10, and 24 h after birth. Prefeeding blood samples taken at 0 h, 10 h, 24 h, 36 h, 4 d, and 7 d were analyzed for serum IgG concentration. Apparent efficiency of IgG absorption in the treatment group was higher compared to that in the control group. Serum IgG concentration of calves in the treatment group was higher for all sampling time points except for 0 h of age. Mean serum IgG concentrations in the treatment and control groups at 36 h of age were 22.3 and 17.7 g/L, respectively. Serum IgG concentration at 36 h of age was positively related with the IgG concentration in the PC in both groups, and it was higher in the treatment group than in the control group at the same IgG concentration in the PC. These results indicate that absorption of ingested IgG by endocytosis as well as a nonselected concentration gradient process of the paracellular pathway of the intestinal epithelium could be enhanced by DFA III supplementation. Supplementation of DFA III in PC containing various IgG concentrations could improve IgG absorption as well as calf serum IgG concentration.

Changes of Serum Calcium Concentration, Frequency of Ruminal Contraction and Feed Intake Soon after Parturition of Dairy Cows Fed Difructose Anhydride III.

Requirements to control the large decrease in serum calcium (Ca) due to parturition and to increase the feed intake soon after parturition have been well accepted in dairy cows. This study was aimed to investigate the feed intake affected by serum Ca concentration with difructose anhydride (DFA) III supplement in dairy cows soon after parturition. Fourteen transition Holstein cows were divided into DFA and control (CONT) groups within 1 to 5 parity variations in each group. Measurement schedule for an individual cow was from 14 d before parturition to 7 d following parturition. The cows in DFA group were supplied 0.2 kg/head/d of DFA III feed containing 40 g of pure DFA III while the cows in CONT group received no DFA III. Other feeding procedures were the same for all cows in both groups. At parturition (d 0), serum Ca concentration sharply declined in both groups (p<0.05). Time interval for recovery from decreased serum Ca to its normal range (>9.0 mg/dL) tended to be faster in DFA group (12 h) than in the CONT group (48 h), but the differences were not significant. Active ruminal contraction was observed in DFA group at following parturition of d 1 (p<0.05), d 3 (p<0.05), and d 5 (p<0.01). Dry matter (DM) intake did not differ between the groups. However, positive correlations were observed between serum Ca concentration and ruminal contraction (p<0.001), and between ruminal contraction and DM intake (p<0.001) during following parturition. According to multiple regression analysis (R(2) = 0.824, p<0.001), the DM intake was positively affected by serum Ca concentration and ruminal contraction. These results suggest that feed intake soon after parturition in dairy cows can be increased by improvement of serum Ca concentration and active ruminal contraction, but DFA III supplementation in this study did not improve the lower serum Ca concentration due to parturition.

Measurement of serum short-chain fatty acid concentrations in cattle after oral administration of difructose anhydride III.

Background and Aim: We previously reported the mitigation effects of difructose anhydride III (DFA III) on mycotoxins, such as zearalenon and sterigmatocystin, based on the urinary concentrations of these molecules in calves. This study was aimed at evaluating the effects of dietary supplementation of DFA III and the fermented status of DFA III in the intestine by comparing serum levels of short-chain fatty acid (SCFAs) in DFA III-supplemented cattle with those in non-supplemented control cattle. Materials and Methods: Serum SCFA concentrations were measured in 30 Japanese Black heifers, aged 9-10 months, from two herds, using gas chromatography on days 0 (before DFA III supplementation), 9, and 14 after DFA III supplementation. Results: A notably different trend was observed for isobutyric acid and enanthic acid, which may reflect the different fermentation status of supplementary DFA III in the intestine. Our results indicate the possibility that this trend observed in the intestinal tract following DFA III administration is associated with changes in the environment of intestinal bacterial flora, which may partially reflect the effects of DFA III supplementation on cattle. Conclusion: Difructose anhydride III supplementation for at least 2 weeks affects the trend of blood SCFA concentrations in cattle. Our results provide evidence supporting the effects of DFA III on the intestinal environment and intestinal barrier function.

Effect of dietary difructose anhydride III supplementation on bone mineral density and calcium metabolism in late-lactation dairy cows.

The aim of the present study was to examine the effect of 28 days of dietary difructose anhydride (DFA) III supplementation on calcium (Ca) metabolism in late-lactation dairy cows. Twenty-four multiparous pregnant Holstein cows were divided into two groups. The DFA group was fed total mixed ration (TMR) supplemented with 40 g of DFA III, and the control group was fed TMR only. The replenishment of bone Ca reserves was evaluated by measuring bone mineral density (BMD) and blood biochemical bone markers. Serum Ca concentrations, urinary Ca-to-creatinine (Cre) (Ca/Cre) ratios, and milk Ca concentrations were also analyzed. The BMD of the 4th caudal vertebra in the DFA group was higher than in the control group on day 28. With respect to bone markers, the ratios of undercarboxylated osteocalcin (ucOC) to osteocalcin (OC) in the DFA group were significantly lower than those in the control group on days 21 and 28. Milk Ca concentrations in the DFA group were also higher than those in the control group on days 14, 21, and 28, whereas serum Ca concentrations and urinary Ca/Cre ratios were unchanged in both groups. These results suggest that dietary supplementation with DFA III increased BMD and decreased serum ucOC/OC ratios in late-lactation dairy cows; this indicates that the replenishment of bone Ca reserves may be enhanced by dietary DFA III supplementation.