Sang-Ho Yoo

Structural and functional characteristics of selected soft wheat starches

ABSTRACT Starches from eight soft wheat samples (two parent lines and six offspring) were isolated; relationships between their structures and properties were examined. Branch chain-length distributions of amylopectins were determined by using high-performance anion exchange chromatography equipped with an amyloglucosidase reactor and a pulsed amperometric detector (HPAEC-ENZ-PAD). Results showed that the average chain length of the eight samples varied at DP 25.6–26.9. Starch samples of lines 02, 60, 63, 95, and 114 consisted of amylopectins with more long chains (DP ≥ 37) and longer average chain length (DP 26.2–26.9) than that of other samples. These starch samples of longer branch chain length displayed higher gelatinization temperatures (55.3–56.5°C) than that of other samples (54.4–54.9°C) and higher peak viscosity (110–131 RVU) and lower pasting temperature (86.3–87.6°C) than others (83–100 RVU and 88.2–88.9°C, respectively). The Mw of amylopectins, determined by using high-performance size exclusi...

Utilization of pectin-enriched materials from apple pomace as a fat replacer in a model food system

Water soluble pectin-enriched materials (PEMs) from apple pomace, were evaluated as a fat replacer in a model food system. When PEM solutions were subjected to steady-shear measurements, shear-thinning behavior was observed. The flow behaviors could be described by the Cross model (R(2)=0.99), and temperature effects were investigated by the Arrhenius equation. The addition of PEMs significantly increased the pasting parameters of wheat flour as measured by a starch pasting rheometer. Gelatinization temperature and enthalpy increased with increasing PEM concentrations. When PEMs were incorporated into cookie formulations in place of shortening (semisolid fat generally used in baked foods) up to 30% by the weight of shortening, the cookie spread diameter was reduced while an increase in the moisture content was observed. Moreover, replacement of shortening with PEMs contributed to a more tender texture and lighter surface color.

Comparative structural analyses of purified glycogen particles from rat liver, human skeletal muscle and commercial preparations.

Glycogen is a cellular energy store that is crucial for whole body energy metabolism, metabolic regulation and exercise performance. To understand glycogen structure we have purified glycogen particles from rat liver and human skeletal muscle tissues and compared their biophysical properties with those found in commercial glycogen preparations. Ultrastructural analysis of commercial liver glycogens fails to reveal the classical alpha-rosette structure but small irregularly shaped particles. In contrast, commercial slipper limpet glycogen consists of beta-particles with similar branching and chain lengths to purified rat liver glycogen together with a tendency to form small alpha-particles, and suggest it should be used as a source of glycogen for all future studies requiring a substitute for mammalian liver glycogen.

Enzymatic synthesis of salicin glycosides through transglycosylation catalyzed by amylosucrases from Deinococcus geothermalis and Neisseria polysaccharea

Amylosucrase (ASase, EC 2.4.1.4) is a member of family 13 of the glycoside hydrolases that catalyze the synthesis of an alpha-(1-->4)-linked glucan polymer from sucrose instead of an expensive activated sugar, such as ADP- or UDP-glucose. Transglycosylation reactions mediated by the ASases of Deinococcus geothermalis (DGAS) and Neisseria polysaccharea (NPAS) were applied to the synthesis of salicin glycosides with sucrose serving as the glucopyranosyl donor and salicin as the acceptor molecule. Two salicin glycoside transfer products were detected by TLC and HPLC analyses. The synthesis of salicin glycosides was very efficient with NPAS with a yield of over 90%. In contrast, DGAS specifically synthesized only one salicin transglycosylation product. The transglycosylation products were identified as alpha-d-glucopyranosyl-(1-->4)-salicin (glucosyl salicin) and alpha-D-glucopyranosyl-(1-->4)-alpha-D-glucopyranosyl-(1-->4)-salicin (maltosyl salicin) by NMR analysis. The ratio between donor and acceptor had a significant effect on the type of product that resulted from the transglycosylation reaction. With more acceptors present in the reaction, more glucosyl salicin and less maltosyl salicin were synthesized.

Enzyme-Synthesized Highly Branched Maltodextrins Have Slow Glucose Generation at the Mucosal α-Glucosidase Level and Are Slowly Digestible In Vivo.

For digestion of starch in humans, α-amylase first hydrolyzes starch molecules to produce α-limit dextrins, followed by complete hydrolysis to glucose by the mucosal α-glucosidases in the small intestine. It is known that α-1,6 linkages in starch are hydrolyzed at a lower rate than are α-1,4 linkages. Here, to create designed slowly digestible carbohydrates, the structure of waxy corn starch (WCS) was modified using a known branching enzyme alone (BE) and an in combination with β-amylase (BA) to increase further the α-1,6 branching ratio. The digestibility of the enzymatically synthesized products was investigated using α-amylase and four recombinant mammalian mucosal α-glucosidases. Enzyme-modified products (BE-WCS and BEBA-WCS) had increased percentage of α-1,6 linkages (WCS: 5.3%, BE-WCS: 7.1%, and BEBA-WCS: 12.9%), decreased weight-average molecular weight (WCS: 1.73×108 Da, BE-WCS: 2.76×105 Da, and BEBA-WCS 1.62×105 Da), and changes in linear chain distributions (WCS: 21.6, BE-WCS: 16.9, BEBA-WCS: 12.2 DPw). Hydrolysis by human pancreatic α-amylase resulted in an increase in the amount of branched α-limit dextrin from 26.8% (WCS) to 56.8% (BEBA-WCS). The α-amylolyzed samples were hydrolyzed by the individual α-glucosidases (100 U) and glucogenesis decreased with all as the branching ratio increased. This is the first report showing that hydrolysis rate of the mammalian mucosal α-glucosidases is limited by the amount of branched α-limit dextrin. When enzyme-treated materials were gavaged to rats, the level of postprandial blood glucose at 60 min from BEBA-WCS was significantly higher than for WCS or BE-WCS. Thus, highly branched glucan structures modified by BE and BA had a comparably slow digesting property both in vitro and in vivo. Such highly branched α-glucans show promise as a food ingredient to control postprandial glucose levels and to attain extended glucose release.

Properties of a Novel Thermostable Glucoamylase from the Hyperthermophilic Archaeon Sulfolobus solfataricus in Relation to Starch Processing

ABSTRACT A gene (ssg) encoding a putative glucoamylase in a hyperthermophilic archaeon, Sulfolobus solfataricus, was cloned and expressed in Escherichia coli, and the properties of the recombinant protein were examined in relation to the glucose production process. The recombinant glucoamylase was extremely thermostable, with an optimal temperature at 90°C. The enzyme was most active in the pH range from 5.5 to 6.0. The enzyme liberated β-d-glucose from the substrate maltotriose, and the substrate preference for maltotriose distinguished this enzyme from fungal glucoamylases. Gel permeation chromatography and sedimentation equilibrium analytical ultracentrifugation analysis revealed that the enzyme exists as a tetramer. The reverse reaction of the glucoamylase from S. solfataricus produced significantly less isomaltose than did that of industrial fungal glucoamylase. The glucoamylase from S. solfataricus has excellent potential for improving industrial starch processing by eliminating the need to adjust both pH and temperature.

Quality improvement of rice noodle restructured with rice protein isolate and transglutaminase

In an effort to improve the properties of rice dough and quality of gluten-free rice noodle, transglutaminase (TGase) and rice protein isolate (RPI) were applied to rice noodle making process. The storage and loss moduli of rice dough increased by TGase treatment, whereas they decreased with RPI supplementation. The combined treatment of RPI+TGase on rice dough fully recovered the reduced moduli caused by RPI only supplementation to control level, and increased most of viscosity parameters of rice noodle in RVA analysis. This additional treatment of TGase also increased development time, and maximum and peak torques of RPI-supplemented rice dough in Mixolab® analysis. Cooking loss and water turbidity of rice noodle decreased by 54.8% and 66.6%, respectively, after TGase+RPI treatment. Scanning electron micrographs showed cracked noodle surface became smoothed with TGase treatment, which was more obvious with RPI+TGase treatment. These results suggest RPI+TGase treatment could improve the quality of rice noodle without the use of gluten-like ingredients.

Characterization of cyanobacterial glycogen isolated from the wild type and from a mutant lacking of branching enzyme.

Cyanobacteria produce glycogen as their primary form of carbohydrate storage. The genomic DNA sequence of Synechocystis sp. PCC6803 indicates that this strain encodes one glycogen-branching enzyme (GBE) and two isoforms of glycogen synthase (GS). To confirm the putative GBE and to demonstrate the presence of only one GBE gene, we generated a mutant lacking the putative GBE gene, sll0158, by replacing it with a kanamycin resistance gene through homologous recombination. GBE in sll0158(-) mutant was eliminated; the mutant strain produced less glucan, equivalent to 48% of that produced by the wild type. In contrast to the wild-type strain that had 74% of the glucan being water-soluble, the mutant had only 14% of the glucan water-soluble. Molecular structures of glucans produced by the mutant and the wild type were characterized by using high-performance size-exclusion and anion-exchange chromatography. The glycogen produced by the wild type displayed a molecular mass of 6.6 x 10(7) daltons (degree of polymerization (DP) 40700) and 10% branch linkages, and the alpha-D-glucan produced by the mutant displayed a molecular mass of 4.7-5.6 x 10(3) daltons (DP 29-35) with slight branch linkages. The results indicated that sll0158 was the major functional GBE gene in Synechocystis sp. PCC6803.

Effects of amylosucrase treatment on molecular structure and digestion resistance of pre-gelatinised rice and barley starches

Structural modification of rice and barley starches with Neisseria polysaccharea amylosucrase (NpAS) was conducted, and relationship between structural characteristics and resistant starch (RS) contents of NpAS-treated starches was investigated. Pre-gelatinised rice and barley starches were treated with NpAS. NpAS-treated starches were characterised with respect to morphology, X-ray diffraction pattern, amylopectin branch-chain distribution, and RS content, and their structural characteristics were correlated to RS contents. Regardless of amylose contents of native starches, NpAS-treated (relative to native) starches possessed lower and higher proportions of shorter (DP 6-12) and intermediate (DP 13-36) amylopectin (AP) branch-chains, respectively. RS contents were higher for NpAS-treated starches relative to native starches, and maximum RS contents were obtained for NpAS-treated starches of waxy rice and barley genotypes. Amylose contents were not associated with RS contents of NpAS-treated starches. However, shorter and intermediate AP branch-chain portions were negatively and positively correlated to RS contents of NpAS-treated starches, respectively.

Molecular Structure of Selected Tuber and Root Starches and Effect of Amylopectin Structure on Their Physical Properties

The objectives of this study were to characterize starches isolated from potato, canna, fern, and kudzu, grown in Hangzhou, China, for potential food and nonfood applications and to gain understandings of the structures and properties of tuber and root starches. Potato and canna starches with B-type X-ray patterns had larger proportions of amylopectin (AP) long branch chains (DP >or=37) than did fern (C-type) and kudzu (C(A)-type) starches. The analysis of Naegeli dextrins suggested that fern and kudzu starches had more branch points, alpha-(1,6)-D-glycosidic linkages, located within the double-helical crystalline lamella than did the B-type starches. Dispersed molecular densities of the C- and C(A)-type APs (11.6-13.5 g/mol/nm(3)) were significantly larger than those of the B-type APs (1.4-6.1 g/mol/nm(3)) in dilute solutions. The larger proportion of the long AP branch chains in the B-type starch granules resulted in greater gelatinization enthalpy changes (Delta H). Retrograded kudzu starch, which had the shortest average chain length (DP 25.1), melted at a lower temperature (37.9 degrees C) than the others. Higher peak viscosities (550-749 RVU at 8%, dsb) of potato starches were attributed to the greater concentrations of phosphate monoesters, longer branch chains, and larger granule sizes compared with other tuber and root starches.