-
Journal of Chromatography. A Jun 2024In recent years, many nucleic acid-based pharmaceuticals have been approved and entered the market, and even a larger number are in late stage clinical trials....
In recent years, many nucleic acid-based pharmaceuticals have been approved and entered the market, and even a larger number are in late stage clinical trials. Conventional oligonucleotides are facing issues in vivo like fast renal clearance and nuclease degradation. Therefore, to increase their stability, phosphorothioation is a frequent modification of therapeutic oligonucleotides (ONs) which also leads to improved binding affinity facilitating cell internalization and intracellular distribution. At the same time, by replacing a phosphodiester linkage with a phosphorothioate group, a phosphorous stereogenic center is generated which causes the formation of R- and S-diastereomers. It increases the structural diversity. For example, with 15 of those phosphorothioate (PS) linkages, 32,768 different diastereomers are expected. Since the phosphorothioate is introduced non-stereoselectively, the molecular complexity of the resultant phosphorothioate ON products is tremendously increased impeding the chromatographic separation in the course of quality control. Since distinct phosphorothioate diastereomers have different bioactivities and pharmacological properties, there is increasing interest in implications of stereoisomerism of phosphorothiate oligonucleotides. From a quality and regulatory viewpoint, batch-to-batch reproducibility of the diastereomer profile may be of significant concern. In order to address this issue, this study investigates the stereoselectivity of LC methods for two phosphorothioate oligonucleotide (PSO) compounds differing in their molecular size and numbers of PS linkages. Diastereoselectivity of ion-pairing reversed-phase liquid chromatography (IP-RPLC), RPLC without ion-pairing agents and LC with chiral polysaccharide-based column were evaluated for model PSOs and an active pharmaceutical ingredient (API) of PSO with trivalent N-acetylgalactosamine (GalNAc) conjugate. Due to the structural complexity of PSOs, the separation power for the diastereomer mixture was increased by using sequential selective comprehensive two-dimensional chromatography with an amylose tris(α-methylbenzylcarbamate)-immobilized chiral stationary phase (CSP) in the first dimension and ion-pair RPLC with ethylammonium acetate in the second dimension. Improved diastereomer selectivity was obtained and a larger number of peaks could be separated.
PubMed: 38879975
DOI: 10.1016/j.chroma.2024.465076 -
Food Research International (Ottawa,... Aug 2024The digestibility of ungelatinized, short-term retrograded and long-term retrograded starch from foxtail millet was investigated and correlated with starch chain length...
The digestibility of ungelatinized, short-term retrograded and long-term retrograded starch from foxtail millet was investigated and correlated with starch chain length distributions (CLDs). Some variations in starch CLDs of different varieties were obtained. Huangjingu and Zhonggu 9 had higher average chain lengths of debranched starch and lower average chain length ratios of amylopectin and amylose than Dajinmiao and Jigu 168. Compared to ungelatinized starch, retrogradation significantly increased the estimated glycemic index (eGI), whereas significantly decreased the resistant starch (RS). In contrast, long-term retrograded starches have lower eGI (93.33-97.37) and higher RS (8.04-14.55%) than short-term retrograded starch. PCA and correlation analysis showed that amylopectin with higher amounts of long chains and longer long chains contributed to reduced digestibility in ungelatinized starch. Both amylose and amylopectin CLDs were important for the digestibility of retrograded starch. This study helps a better understanding of the interaction of starch CLDs and digestibility during retrogradation.
Topics: Setaria Plant; Digestion; Starch; Amylopectin; Amylose; Glycemic Index
PubMed: 38876595
DOI: 10.1016/j.foodres.2024.114563 -
Food Chemistry Jun 2024Multi-scale structures were investigated to understand starch digestibility of instant rice. A wide range of maximum starch digested ratio, up to about 20%, was observed...
Multi-scale structures were investigated to understand starch digestibility of instant rice. A wide range of maximum starch digested ratio, up to about 20%, was observed among instant rice prepared from different rice varieties. Instant rice with a smooth and densely packed cross-section showed slower starch digestibility than those with a porous and loosely packed structure. All samples displayed B + V type crystallinity, with V-type crystallinity negatively correlating with maximum starch digested percentage. After digestion, starch chain-length distributions were significantly altered: rapidly digested starch comprised long amylose and short amylopectin chains, while slowly digested starch comprised chains with a peak degree of polymerization (DP) around 130. These results indicate that instant rice with a compact microstructure, high V-type crystallinity, and DP 130 fractions during digestion can reduce starch digestibility. This study provides insights for food industry to develop instant rice products with slow starch digestibility, potentially improving human health.
PubMed: 38876074
DOI: 10.1016/j.foodchem.2024.140074 -
Rice (New York, N.Y.) Jun 2024Improving rice quality remains a crucial breeding objective, second only to enhancing yield, yet progress in quality improvement lags behind yield. The high temperature...
Improving rice quality remains a crucial breeding objective, second only to enhancing yield, yet progress in quality improvement lags behind yield. The high temperature and ripening conditions in Southern China often result in poor rice quality, impacting hybrid rice production and utilization. Therefore, to address this challenge, analyzing the molecular basis of high-quality traits is essential for molecular design breeding of high-quality hybrid rice varieties. In this study, we investigated the molecular basis of grain shape, amylose content, gel consistency, gelatinization temperature, and aroma, which influence rice quality. We discovered that quality related alleles gs3, GW7, gw8, chalk5, Wx, ALK, and fgr can enhance rice quality when applied in breeding programs. Polymerization of gs3, GW7, gw8, and chalk5 genes improves rice appearance quality. The gs3 and GW7 allele polymerization increasing the grain's length-width ratio, adding the aggregation of gw8 allele can further reducing grain width. The chalk5 gene regulates low chalkiness, but low correlation to chalkiness was exhibited with grain widths below 2.0 mm, with minimal differences between Chalk5 and chalk5 alleles. Enhancing rice cooking and eating quality is achieved through Wx and ALK gene polymerization, while introducing the fgr(E7) gene significantly improved rice aroma. Using molecular marker-assisted technology, we aggregated these genes to develop a batch of indica hybrid rice parents with improved rice quality are obtained. Cross-combining these enhanced parents can generate new, high-quality hybrid rice varieties suitable for cultivation in Southern China. Therefore, our findings contribute to a molecular breeding model for grain quality improvement in high-quality indica hybrid rice. This study, along with others, highlights the potential of molecular design breeding for enhancing complex traits, particularly rice grain quality.
PubMed: 38874692
DOI: 10.1186/s12284-024-00719-7 -
TAG. Theoretical and Applied Genetics.... Jun 2024Integrated linkage and association analysis revealed genetic basis across multiple environments. The genes Zm00001d003102 and Zm00001d015905 were further verified to...
Integrated linkage and association analysis revealed genetic basis across multiple environments. The genes Zm00001d003102 and Zm00001d015905 were further verified to influence amylose content using gene-based association study. Maize kernel amylose is an important source of human food and industrial raw material. However, the genetic basis underlying maize amylose content is still obscure. Herein, we used an intermated B73 × Mo17 (IBM) Syn10 doubled haploid population composed of 222 lines and a germplasm set including 305 inbred lines to uncover the genetic control for amylose content under four environments. Linkage mapping detected 16 unique QTL, among which four were individually repeatedly identified across multiple environments. Genome-wide association study revealed 17 significant (P = 2.24E-06) single-nucleotide polymorphisms, of which two (SYN19568 and PZE-105090500) were located in the intervals of the mapped QTL (qAC2 and qAC5-3), respectively. According to the two population co-localized loci, 20 genes were confirmed as the candidate genes for amylose content. Gene-based association analysis indicated that the variants in Zm00001d003102 (Beta-16-galactosyltransferase GALT29A) and Zm00001d015905 (Sugar transporter 4a) affected amylose content across multi-environment. Tissue expression analysis showed that the two genes were specifically highly expressed in the ear and stem, respectively, suggesting that they might participate in sugar transport from source to sink organs. Our study provides valuable genetic information for breeding maize varieties with high amylose.
Topics: Zea mays; Amylose; Chromosome Mapping; Quantitative Trait Loci; Polymorphism, Single Nucleotide; Genome-Wide Association Study; Phenotype; Genetic Linkage; Genes, Plant; Genotype; Genetic Association Studies
PubMed: 38872054
DOI: 10.1007/s00122-024-04666-1 -
PloS One 2024BRRI31R is one of the Bangladesh's most promising restorer lines due to its abundant pollen producing capacity, strong restoring ability, good combining ability, high...
Pyramiding of bacterial blight resistance genes into promising restorer BRRI31R line through marker-assisted backcross breeding and evaluation of agro-morphological and physiochemical characteristics of developed resistant restorer lines.
BRRI31R is one of the Bangladesh's most promising restorer lines due to its abundant pollen producing capacity, strong restoring ability, good combining ability, high outcrossing rate and genetically diverse from cytoplasmic male sterile (CMS) line. But the drawback of this line is that it is highly susceptible to bacterial blight (BB) disease of rice caused by Xanthomonas oryzae pv. oryzae. The present study highlighted the pyramiding of effective BB resistance genes (xa5, xa13 and Xa21) into the background of BRRI31R, through marker-assisted backcrossing (MABC). Backcross progenies were confirmed and advanced based on the foreground selection of target genes. Pyramided lines were used for pathogenicity test against five Bangladeshi Xanthomonas oryzae (BXo) races (BXo93, BXo220, BXo822, BXo826, BXo887) and confirmed the dominant fertility restore genes, Rf3 and Rf4 and further validated against SNP markers for more confirmation of target resistance genes. All pyramided restorer lines consisted of Xa4 (in built), xa5, xa13, Xa21, and Chalk5 with two fertility restorer genes, Rf3, Rf4. and these restorer lines showed intermediate amylose content (<25%). Restorer lines BRRI31R-MASP3 and BRRI31R-MASP4 showed high levels of resistance against five virulent BXo races and SNP genotyping revealed that these lines also contained a blast resistance gene Pita races. Gene pyramided restorer lines, BRRI31R-MASP3 and BRRI31R-MASP4 can directly be used as a male parent for the development of new BB resistant hybrid rice variety or could be used as a replacement of restorer line of BRRI hybrid dhan5 and 7 to enhance the quality of hybrid seeds as well as rice production in Bangladesh.
Topics: Disease Resistance; Plant Diseases; Xanthomonas; Oryza; Plant Breeding; Genes, Plant; Genetic Markers; Crosses, Genetic
PubMed: 38865348
DOI: 10.1371/journal.pone.0301342 -
Current Research in Food Science 2024The objective of this work was to completely replace margarine with peanut diacylglycerol oil/ethyl cellulose-glycerol monostearate oleogel (DEC/GMS) oleogel, and...
The objective of this work was to completely replace margarine with peanut diacylglycerol oil/ethyl cellulose-glycerol monostearate oleogel (DEC/GMS) oleogel, and evaluate its effect on starch digestibility of cakes. The in vitro digestibility analysis demonstrated that the DEC/GMS-6 cake exhibited a 26.36% increase in slowly digestible starch (SDS) and resistant starch (RS) contents, compared to cakes formulated with margarine. The increased SDS and RS contents might mainly be due to the hydrophobic nature of OSA-wheat flour, which could promote the formation of lipid-amylose complexes with GMS and peanut diacylglycerol oil. XRD pattern suggested that the presence of GMS in DEC-based oleogels facilitated the formation of lipid-amylose complexes. The DSC analysis revealed that the addition of GMS resulted in a significant increase in gelatinization enthalpy, rising from 249.7 to 551.9 J/g, which indicates an improved resistance to gelatinization. The FTIR spectra indicated that the combination of GMS could enhance the hydrogen bonding forces and short-range ordered structure in DEC-based cakes. The rheological analysis revealed that an increase in GMS concentration resulted in enhanced viscoelasticity of DEC-based cake compared to TEC-based cakes. The DEC-based cakes exhibited a more satisfactory texture profile and higher overall acceptability than those of TEC-based cakes. Overall, these findings demonstrated that the utilization of DEC-based oleogel presented a viable alternative to commercial margarine in the development of cakes with reduced starch digestibility.
PubMed: 38860263
DOI: 10.1016/j.crfs.2024.100770 -
Frontiers in Nutrition 2024Research on the physicochemical properties of rice-derived endo-sperm high resistant starch (RS) with low amylose content (AC) is limited. In this study, we evaluated...
Research on the physicochemical properties of rice-derived endo-sperm high resistant starch (RS) with low amylose content (AC) is limited. In this study, we evaluated the physicochemical characteristics of such a starch variety and revealed that the starch granules exhibit a smoother, more refined surface with distinct edges, increased compactness, higher order of surface, and fewer cavities compared to those of a low RS rice variety. The starch crystal was classified as an A-type, which may be connected to the high amylose-lipid complex content. The branched internal long chains (B2 + B3) were abundant, allowing for easy entanglement with other molecular chains and a compact structure. Differential scanning calorimetry revealed the need for high temperature and energy to disrupt the double helix structure within the crystallization region of starch. Furthermore, starch viscosity analysis revealed a high cold paste viscosity, consistency, and setback value, with recrystallization yielding a stable structure, increased viscosity, and enhanced hydrolysis resistance to enzymes.
PubMed: 38860156
DOI: 10.3389/fnut.2024.1413923 -
Carbohydrate Polymers Sep 2024The objective of this work was to study the effects of heat-moisture treatment (HMT) of freshly harvested mature high-amylose maize (HAM) kernels on its starch...
The objective of this work was to study the effects of heat-moisture treatment (HMT) of freshly harvested mature high-amylose maize (HAM) kernels on its starch structure, properties, and digestibility. Freshly harvested HAM kernels were sealed in Pyrex glass bottles and treated at 80 °C, 100 °C, or 120 °C. HMT of HAM kernels had no impact on its starch X-ray diffraction pattern but increased the relative crystallinity. This result together with the increased starch gelatinization temperatures and enthalpy change indicated starch molecules reorganization forming long-chain double-helical crystalline structure during HMT of HAM kernels. The aggregation of starch granules were observed after HMT, indicating interaction of starch granules and other components. This interaction and the high-temperature crystalline structure led to reductions in the starch digestibility, swelling power, solubility, and pasting viscosity of the HAM flours. Some starch granules remained intact and showed strong birefringence after the HAM flours were precooked at 100 °C for 20 min and followed by enzymatic hydrolysis, and the amount of undigested starch granules increased with increasing HMT temperatures. This result further supported that HMT of HAM kernels with high moisture level could increase the starch thermal stability and enzymatic resistance.
Topics: Zea mays; Amylose; Hot Temperature; Starch; Hydrolysis; Viscosity; Solubility; Water; X-Ray Diffraction; Flour
PubMed: 38858024
DOI: 10.1016/j.carbpol.2024.122303 -
Carbohydrate Polymers Sep 2024The recently characterized Limosilactobacillus reuteri N1 GtfB (LrN1 GtfB) from glycoside hydrolase family 70 is a novel 4,6-α-glucanotransferase acting on...
The recently characterized Limosilactobacillus reuteri N1 GtfB (LrN1 GtfB) from glycoside hydrolase family 70 is a novel 4,6-α-glucanotransferase acting on starch/maltooligosaccharides with high enzyme activity and soluble protein yield (in heterogenous system). In this study, the influence of the treatment by LrN1 GtfB on the fine structure and functional characteristics of three maize starches were furtherly investigated and elucidated. Due to the treatment of LrN1 GtfB, the starch molecules were transformed into reuterans containing linear and branched (α1 → 6) linkages with notably smaller molecular weight and shorter chain length. Moreover, the (α1 → 6) linkage ratios in the GtfB-modified high-amylose maize starch (GHMS)/normal maize starch (GNMS)/waxy maize starch (GWMS) increased by 18.3 %/12.6 %/9.0 % as compared to their corresponding controls. In vitro digestibility experiment revealed that the resistant starch content of GHMS, GNMS and GWMS increased by 16 %, 18 % and 25 % as compared to the starch substrates. Furthermore, the butyric acid yielded from GHMS, GNMS and GWMS in the in vitro fermentation experiments were 1.4, 1.5 and 1.4 times higher than those of commercial galactose oligosaccharides. These results indicated that the highly-branched short-clustered reuteran synthesized by LrN1 GtfB might serve as novel potential prebiotics, and provide insights for the synthesis of promising prebiotic dietary fiber from starch.
Topics: Zea mays; Limosilactobacillus reuteri; Starch; Prebiotics; Oligosaccharides
PubMed: 38858018
DOI: 10.1016/j.carbpol.2024.122249