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Eleven oligosaccharides were isolated from sugar beet molasses using carbon-Celite column chromatography and HPLC. The constituent sugars and linkage positions were determined using methylation analysis, MALDI-TOF-MS, and NMR measurements. The configurations of isolated oligosaccharides were confirmed based on detailed NMR analysis. Based on our results, three of the 11 oligosaccharides were novel., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Four oligosaccharides containing a fructopyranosyl residue have been found from fermented beverage of plant extract and isolated from the beverage using carbon-Celite column chromatography and preparative high performance liquid chromatography. Structure confirmation of the saccharides was provided by methylation analysis, MALDI-TOF-MS and NMR measurements. These saccharides were identified as oligosaccharides of fructopyranoside series; beta-D-fructopyranosyl-(2-->6)-D-fructofuranose (1), beta-D-fructopyranosyl-(2-->1)-D-fructopyranose (2), beta-D-fructopyranosyl-(2-->1)-beta-D-fructofuranosyl-(2<-->1)-alpha-D-glucopyranoside (3), and beta-D-fructopyranosyl-(2-->6)-alpha-D-glucopyranosyl-(1<-->2)-beta-D-fructofuranoside (4). Saccharides 3 and 4 among novel saccharides 1, 3, and 4 were named 'pyrano-1-kestose (pyrano-isokestose)' and 'pyrano-neokestose', respectively., ((c) 2009 Elsevier Ltd. All rights reserved.)

An extract from 50 kinds of fruits and vegetables was fermented to produce a new beverage. Natural fermentation of the extract was carried out mainly by lactic acid bacteria (Leuconostoc spp.) and yeast (Zygosaccharomyces spp. and Pichia spp.). Two new saccharides were found in this fermented beverage. The saccharides were isolated using carbon-Celite column chromatography and preparative high performance liquid chromatography. Gas liquid chromatography analysis of methylated derivatives as well as MALDI-TOF MS and NMR measurements were used for structural confirmation. The (1)H and (13)C NMR signals of each saccharide were assigned using 2D-NMR including COSY, HSQC, HSQC-TOCSY, CH(2)-HSQC-TOCSY, and CT-HMBC experiments. The saccharides were identified as beta-D-fructopyranosyl-(2-->6)-beta-D-glucopyranosyl-(1-->3)-D-glucopyranose and beta-D-fructopyranosyl-(2-->6)-[beta-D-glucopyranosyl-(1-->3)]-D-glucopyranose.

Fermented beverage of plant extract was prepared from about 50 kinds of vegetables and fruits. Natural fermentation was carried out mainly by lactic acid bacteria (Leuconostoc spp.) and yeast (Zygosaccharomyces spp. and Pichia spp.). Three kinds of saccharides have been found in this beverage and produced by fermentation. The saccharides isolated from the beverage using carbon-Celite column chromatography and preparative HPLC, were identified as a new saccharide, beta-d-fructopyranosyl-(2-->6)-d-glucopyranose, laminaribiose and maltose by examination of constituted sugars, GLC and GC-MS analyses of methyl derivatives and MALDI-TOF-MS and NMR measurements of the saccharides.

The objective of this study was to report on the variation of DP 6 isomers (6b, 6c + 6d1 + 6d2), 7a, 8, and 9-12 fructooligosaccharides (FOS) and their hydrolysis parameters [percent hydrolysis, consumption rate, hydrolysis rate constant (k(obsd)), and half-life time (t(1/2))] in onion bulb tissues stored for 6 months at 10, 15, or 20 degrees C. The hydrolysis of DP 6 isomers, 7a, and 8 ranged from 74 to 85%, whereas that of DP 9-12 averaged 86%. The consumption rate of 6b, 6c + 6d1 + 6d2, 7a, 8, and -12 averaged 25, 58, 38, 26, and 48 microg/g of fresh weight per week, respectively. The k(obsd) showed large variation from 53 x 10(-3) week(-1) (lowest value) to 92 x 10(-3) week(-1) (highest value), whereas the half-life t(1/2) of the different FOS ranged between 7.5 and 13.1 weeks. DP 6b isomers increased during the first month, and then the content decreased sharply during the second month at 20 degrees C but remained stable during the last 4 months, whereas at 10 and 15 degrees C, 6b decreased progressively from the first to the six month. In contrast, DP 6c + 6d1 + 6d2 decreased abruptly within the first 3 months at 10, 15, and 20 degrees C; however, during the last 3 months they remained stable, ranging between 0.32 and 0.39. Variation of DP 7a, 8, and 9-12 FOS was close to that of DP 6 FOS isomers. DP 7a increased slightly during the first month, and afterward 7a started decreasing progressively during the last 5 months. DP 8 FOS showed a similar pattern independent of temperature regime: they increased slightly within the first month, but from the second month, DP 8 began decreasing progressively and continued decreasing to the sixth month. DP 9-12 FOS also varied similarly to DP 6c + 6d1 + 6d2. They decreased sharply within the first 2 months, and during the last 4 months, they continued to decrease slowly. Surprisingly, these variations occurred independently of temperature regimes and were affected only by storage duration. It was concluded that highly polymerized FOS are preferably hydrolyzed rather than low DP FOS isomers because they have a relatively high content of fructosyl end chains.

A fructosyltransferase that transfers the terminal (2 --> 1)-beta-linked D-fructosyl group of fructo-oligosaccharides (1(F)(1-beta-D-fructofuranosyl)(n) sucrose, n >/= 1) to HO-6 of the glucosyl residue and HO-1 of the fructosyl residue of similar saccharides (1(F)(1-beta-D-fructofuranosyl)(m) sucrose, m >/= 0) has been purified from an extract of the bulbs of onion (Allium cepa). Successive column chromatography using DEAE-Sepharose CL-6B, Toyopearl HW65, Toyopearl HW55, DEAE-Sepharose CL-6B (2nd time), Sephadex G-100, Concanavalin A Sepharose, and Toyopearl HW-65 (2nd time) were applied for protein purification. The general properties of the enzyme, were as follows: molecular masses of 66 kDa (gel filtration chromatography), and of 52 kDa and 25 kDa (SDS-PAGE); optimum pH of c. 5.68, stable at 20-40 degrees C for 15 min; stable in a range of pH 5.30-6.31 at 30 degrees C for 30 min, inhibited by Hg(2+), Ag(+), p-chloromercuribenzoic acid (p-CMB) and sodium dodecyl sulfate (SDS), activated by sodium deoxycholate, Triton X-100 and Tween-80. The amino acid sequence of the N-terminus moiety of the 52-kDa polypeptide was ADNEFPWTNDMLAWQRCGFHFRTVRNYMNDPSGPMYYKGWYHLFYQHNKDFAYXG and the amino acid sequence from the N-terminus of the 25-kDa polypeptide was ADVGYXCSTSGGAATRGTLGPFGLL VLANQDLTENTATYFYVSKGTDGALRTHFCQDET. The enzyme tentatively classified as fructan: fructan 6(G)-fructosyltransferase (6G-FFT). The enzyme is proposed to play an important role in the synthesis of inulin and inulinneo-series fructo-oligosaccharides in onion bulbs.

Five novel oligosaccharides (tetra-, penta- and hexa-saccharides) were synthesized by glucosyltransfer from beta-D-glucose 1-phosphate to isokestose (O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)-alpha-D-glucopyranoside) or nystose (O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)-O-beta-D-fructofuranosyl-(2-->1)-alpha-D-glucopyranoside) using Thermoanaerobacter brockii kojibiose phosphorylase. The oligosaccharides were identified as 2(2-alpha-D-glucopyranosyl)(m)isokestose; [O-alpha-D-glucopyranosyl-(1-->2)](m)-O-[beta-D-fructofuranosyl-(2-->1)](2)-alpha-D-glucopyranoside: m=1, 2, and 3, and 2(2-alpha-D-glucopyranosyl)(n)nystose; [O-alpha-D-glucopyranosyl-(1-->2)](n)-O-[beta-D-fructofuranosyl-(2-->1)](3)-alpha-D-glucopyranoside: n=1 and 2 using gas liquid chromatography analysis of the methyl derivatives, and MALDI-TOF-MS and NMR measurements of the newly formed oligosaccharides. 1H, 13C NMR signals of each saccharide were assigned using 2D-NMR techniques, including COSY, HSQC, HSQC-TOCSY, HMBC, CH(2)-selected E-HSQC, and CH(2)-selected E-HSQC-TOCSY.

Two novel oligosaccharides, tetra-and penta-saccharides were synthesized by fructosyl transfer from 1-kestose to 4G-beta-D-galactopyranosylsucrose with a purified 1F-fructosyltransferase of asparagus roots and identified as 1F-beta-D-fructofuranosyl-4G-beta-D-galactopyranosylsucrose, O-beta-D-fructofuranosyl-(2-->1)-beta-D-fructofuranosyl-O-[beta-D-galactopyranosyl-(1-->4)]-alpha-D-glucopyranoside and 1F(1-beta-D-fructofuranosyl)2-4G-beta-D-galactopyranosylsucrose, [O-beta-D-fructofuranosyl-(2-->1)]2-beta-D-fructofuranosyl-O-[beta-D-galactopyranosyl-(1-->4)]-alpha-D-glucopyranoside, respectively. Both oligosaccharides were scarcely hydrolyzed by carbohydrase from rat small intestine. Human intestinal bacterial growth by 1F-beta-D-fructofuranosyl-4G-beta-D-galactopyranosylsucrose was compared with that by the tetrasaccharides, stachyose and nystose. Bifidobacteria utilized 1F-beta-D-fructofuranosyl-4G-beta-D-galactopyranosylsucrose to the same extent as stachyose or nystose. On the other hand, the unfavorable bacteria, Clostridium perfringens, Escherichia coli and Enterococcusfaecalis, that produce mutagenic substances did not use the synthetic oligosaccharide.

Abstract: Burdock roots were stored for six weeks at three temperatures, 0, 15 and 20°C, and activity of sucrose hydrolase (SH), 1-kestose hydrolase (1-KH), inulin hydrolase (InH), sucrose:sucrose 1-fructosyltransferase (1-SST) and fructan:fructan 1-fructoslytransferase (1-FFT) were assessed. Simultaneously, total fructooligosaccharides (FOS), total inulooligosaccharides (IOS) and inulin were also assessed during storage. During storage, SH showed an irregular variation at 15 and 20°C, and was significantly higher at 0°C showing a continuous increase during the storage period. 1-KH decreased progressively at 15 and 20°C, but showed a sharp increase at 0°C after two weeks and decreased afterwards. InH showed a different although more regular pattern by decreasing progressively at 0, 15 and 20°C. However, the decrease was more significant at 15 and 20°C during the first two weeks, while at 0°C the decrease was significant after four weeks storage. 1-SST and 1-FFT activities decreased progressively in a pseudo-linear regression during storage, and showed similar patterns. At 15 and 20°C, total FOS increased during the first three weeks then decreased, while at 0°C FOS increased progressively during storage. Total IOS increased during storage, however, this increase was much higher at 0°C than that observed at 15 and 20°C. Inulin content decreased during storage and final content was lower at 20°C. The inulin to total FOS plus total IOS ratio decreased sharply during the first two weeks of storage, then progressively during the last four weeks, while the ratio of hydrolyzing to synthesizing activities was quite stable at 15 and 20°C, but showed a peak at 0°C after two weeks. The results suggest that carbohydrate metabolism in stored burdock depends partly on temperature and other physiological factors. [Copyright &y& Elsevier]