Effects of vitreousness and particle size of maize grain on ruminal and intestinal in sacco degradation of dry matter,starch and nitrogen

We assessed the effects of vitreousness and particle size of maize grain on ruminal and intestinal in sacco degradation of dry matter, starch and nitrogen. Six maize grain (Zea mays) genotypes characterized by differing vitreousness (proportion of vitreous in total endosperm) were ground (3-mm screen; Gr, ground particles, mean particle size (MPS): 526 μm) and cracked with a roller mill using two gap width settings (CS, cracked small particles, MPS: 1360 μm; CL, cracked large particles, MPS: 2380 μm). The ruminal and intestinal in sacco degradation of dry matter, starch and nitrogen was measured on three dry Holstein cows, fitted with rumen, proximal duodenum and terminal ileum cannulas, fed maize silage ad libitum twice daily. The ruminal starch degradability and intestinal digestibility differed among genotypes (P<0.001) and decreased as particle size increased (P<0.001). For the same particle size, starch ruminal degradability decreased (P<0.05) and intestinal digestibility decreased (P<0.002) with vitreousness. Particle size and vitreousness of maize grain are efficient factors for manipulating the amount of starch escaping rumen degradation, but may be limiting for the amount of starch digested in the small intestine.     

Particle morphology characterization and manipulation in biomass slurries and the effect on rheological properties and enzymatic conversion

An improved understanding of how particle size distribution relates to enzymatic hydrolysis performance and rheological properties could enable enhanced biochemical conversion of lignocellulosic feedstocks. Particle size distribution can change as a result of either physical or chemical manipulation of a biomass sample. In this study, we employed image processing techniques to measure slurry particle size distribution and validated the results by showing that they are comparable to those from laser diffraction and sieving. Particle size and chemical changes of biomass slurries were manipulated independently and the resulting yield stress and enzymatic digestibility of slurries with different size distributions were measured. Interestingly, reducing particle size by mechanical means from about 1 mm to 100 μm did not reduce the yield stress of the slurries over a broad range of concentrations or increase the digestibility of the biomass over the range of size reduction studied here. This is in stark contrast to the increase in digestibility and decrease in yield stress when particle size is reduced by dilute-acid pretreatment over similar size ranges.     

Expression of a fungal glucoamylase in transgenic rice seeds

Glucoamylase, which catalyses the hydrolysis of the α-1,4 glycosidic bonds of starch, is an important industrial enzyme used in starch enzymatic saccharification. In this study, a glucoamylase gene from Aspergillus awamori, under the control of the promoter of seed storage protein Gt1, was introduced into rice by Agrobacterium-mediated transformation. Significant glucoamylase activity was detected specifically in the seeds but not other tissues of the transgenic rice lines. The highest enzymatic activity was found in the transgenic line Bg17-2, which was estimated to have about 500 units per gram of seeds (one unit is defined as the amount of enzyme that produces 1 μmol of reducing sugar in 1 min at 60 °C using soluble starch as substrate). The optimum pH for the activity of the rice produced enzyme is 5.0–5.5, and the optimum temperature is around 60 °C. One part of this transgenic glucoamylase rice seed flour fully converted 25 parts of corn starch pre-liquefied by an α-amylase also produced by a transgenic rice into glucose in 16 h incubation. This study suggests that this hydrolysis enzyme may substitute commercial fermentation enzymes for industrial starch conversion.     

Expression of a bi-functional and thermostable amylopullulanase in transgenic rice seeds leads to autohydrolysis and altered composition of starch

Overexpression of bacterial-derived starch metabolic enzymes in plant starch storage organs represents a valuable strategy for improving starch quality, bioprocessing and nutritional value. Transgenic rice seeds producing a thermostable and bifunctional starch hydrolase, amylopullulanase (APU) from Thermoanaerobacter ethanolicus 39E, were generated. Starch in these seeds could be hydrolyzed with optimal temperatures between 85 and 95 °C, which resulted in complete conversion of starch into soluble sugars and production of protein-enriched flour within a few hours. By expressing various levels of APU, rice seeds containing reduced amounts of amylose, which is an important factor affecting starch quality, were obtained without a significant impact on grain yield. Elevation in granule-bound pullulanase activity correlates with the reduction of amylose in developing APU-containing rice seeds. APU was found to be localized within amyloplasts and in cell walls, which could be the result of overexpression of APU with a signal peptide. This study establishes novel approaches to alter starch properties, accelerate bioprocessing of starch and production of protein-enriched flour from rice seeds, and could significantly impact the industrial and food uses of cereals.     

The role of cotyledon cell structure during in vitro digestion of starch in navy beans

Studies on the physico-chemical, microstructural characteristics and in vitro (under simulated gastric and small intestine conditions) starch digestibility of navy beans were carried out. The microstructure of raw and cooked beans observed through scanning electron microscopy (SEM) showed the presence of hexagonal or angular shaped cotyledon cells (50-100 μm size) containing starch granules with a size ranging between 10 and 50 μm. The extent of starch hydrolysis (%) after 120 min of in vitro gastro-intestinal digestion differed between whole navy beans (∼60%) and milled bean flour and bean starch (85-90%) after they were cooked under similar conditions. Starch hydrolysis (%) increased significantly when the cotyledon cells in the cooked whole navy beans were disrupted using high pressure treatment (French press). The storage of freshly cooked whole beans resulted in a lower (40-45%) starch hydrolysis whereas re-heating increased the same to 70-80% during in vitro small intestinal digestion. The SEM pictures of cooked navy bean digesta after different intervals of in vitro gastric and small intestinal digestion showed that the cotyledon cell structure is maintained well throughout the digestion period. However cotyledon cells appear shrunken and developed wrinkles during in vitro digestion. Particle size analysis of cooked bean paste taken before and after the in vitro gastro-intestinal digestion showed similar particle size distributions.     

Fermentation of wheat grain by Schwanniomyces castellii

Fermentation of flour and wheat grain, without enzymatic hydrolysis has been demonstrated using a strain of Schwanniomyces castellii. Ethanol production yields were fairly good although starch was not totally degraded. The composition of the fermentation residues was compared to that of distillers dried grains.     

Enzymatic production of glucose from different qualities of grain sorghum and application of ultrasound to enhance the yield

The objective of the present work was to add value to three different qualities of grain sorghum namely normal healthy, germinated, and blackened through production of glucose, and to intensify glucose production (yield) by means of ultrasound treatment. Liquefaction (using Bacillus licheniformis α-amylase) and saccharification (using amyloglucosidase) processes were optimized with use of normal sorghum flour as a starting material for the production of glucose. The effect of ultrasound treatment on the sorghum slurry prior to liquefaction was studied on the processes of liquefaction and saccharification under optimized conditions. Due to ultrasound treatment, liquefact DE increased by 10-25% depending upon sonication time and the intensity. Ultrasound treatment of 1 min at 100% amplitude was found to decrease the average particle size of the slurry from 302 μm to 115 μm, which resulted in an increased percentage of saccharification by about 8%. The reason for the increase in the percentage of saccharification was attributed to the availability of additional starch for hydrolysis due to ultrasound-assisted disruption of the protein matrix (surrounding starch granules) and the amylose-lipid complex. Integration of ultrasound treatment in the state of art of the production of glucose from dry-milled sorghum and its possible subsequent use in the bioethanol production may improve the overall economics of the process.     

Physicochemical properties and in vitro digestibility of flour and starch from pea (Pisum sativum L.) cultivars

Flours and isolated starches from three different cultivars (1544-8, 1658-11 and 1760-8) of pea grown under identical environmental conditions were evaluated for their physicochemical properties and in vitro digestibility. The protein content, total starch content and apparent amylose content of pea flour ranged from 24.4 to 26.3%, 48.8 to 50.2%, and 13.9 to 16.7%, respectively. In pea starches, the 1760-8 showed higher apparent amylose content and total starch content than the other cultivars. Pea starch granules were irregularly shaped, ranging from oval to round with a smooth surface. All pea starches showed C-type X-ray diffraction pattern with relative crystallinity ranging between 23.7 and 24.7%. Pea starch had only a single endothermic transition (12.1-14.2 J/g) in the DSC thermogram, whereas pea flour showed two separate endothermic transitions corresponding to starch gelatinization (4.54-4.71 J/g) and disruption of the amylose-lipid complex (0.36-0.78 J/g). In pea cultivars, the 1760-8 had significantly higher setback and final viscosity than the other cultivars in both pea flour (672 and 1170 cP, respectively) and isolated starch (2901 and 4811 cP). The average branch chain length of pea starches ranged from 20.1 to 20.3. The 1760-8 displayed a larger proportion of short branch chains, DP (degree of polymerization) 6-12 (21.1%), and a smaller proportion of long branch chains, DP ≥ 37 (8.4%). The RDS, SDS and RS contents of pea flour ranged from 23.7 to 24.1%, 11.3 to 12.8%, and 13.2 to 14.8%, respectively. In pea starches, the 1760-8 showed a lower RDS content but higher SDS and RS contents. The expected glycemic index (eGI), based on the hydrolysis index, ranged from 36.9 to 37.7 and 69.8 to 70.7 for pea flour and isolated pea starch, respectively.     

Comparison of the rates of enzymatic hydrolysis of pretreated rice straw and bagasse with celluloses

The rates of enzymatic hydrolysis of pretreated rice straw and bagasse have been studied and compared with the hydrolysis rates of microcrystalline cellulose powder (MCCP) and Solka Floc. The effects of particle size reduction and enzyme loading on the rates of hydrolysis of rice straw and bagasse were also studied. It was found that the rates of hydrolysis of pretreated rice straw and bagasse are much higher than that of MCCP and Solka Floc. For both rice straw and bagasse, particle size reduction had very little effect in enhancing the rate of hydrolysis. Lignin present at <10% did not seem to hinder the accessibility of the enzyme to the cellulose surface. An enzyme loading > 40 Ug^?1 had no effect on the hydrolysis rate of rice straw or bagasse.     

Cost reduction and feedstock diversity for sulfuric acid-free ethanol cooking of lignocellulosic biomass as a pretreatment to enzymatic saccharification

We have previously demonstrated that a sulfuric acid-free ethanol (EtOH) cooking treatment enhances the enzymatic digestibility of eucalyptus wood and bagasse flour. In the present study, a reconfigured process that achieves similar performance was developed by identifying possible cost-competitive pretreatments that provide high cellulose-to-glucose conversion during subsequent enzymatic hydrolysis. The series of reconfigurations reduced EtOH usage in the pretreatment by more than 80% in comparison with our previous research. Higher initial pressures and intensive size reduction of the starting material are not required. The reconfigured process was applied to rice straw and Douglas fir, in order to confirm the feasibility of feedstock diversity.     

Ozone gas affects physical and chemical properties of wheat (Triticum aestivum L.) starch

In this experiment, bread wheat flour and isolated wheat starch were treated with ozone gas (1,500 mg/kg at 2.5 L/min) for 45 min and 30 min, respectively. Starch was isolated from treated flour. Ozone treated starch and starch isolated from ozone treated flour had similar chemical and physical properties. Chemical analysis of starch isolates indicated depolymerization of high molecular weight amylopectins; with a subsequent increase in low molecular weight starch polymers as a result of starch hydrolysis. Ozone treatment resulted in elevated levels of carboxylic groups and decreased total carbohydrate content in amylopectin fractions. ¹H NMR results indicated formation of a keto group [(1→4)-3 keto] at the H-2 terminal (proton at C-2 position) and β-glucuronic acid at the H-1 terminal (proton at C-1 position). DSC transition temperatures and change in enthalpy were not affected by ozone treatment. Increased swelling power and RVA breakdown were observed in starch from ozone treated samples.     

Studies on the Cereal Starches

Significant correlations among amylograph characteristics or other relating properties of rice starch and flour were recognized by orderly rearrangement of previously obtained data. Linear regression of peak viscosity on breakdown was available for detecting a deteriorative damage of rice starch and grain. Alpha-amylase in white rice did not have close relation with peak viscosity of the flour. The ratio of breakdown to consistency and the corrected peak viscosity calculating the coincidence of the slurry concentration were significantly correlated with the starch iodine blue value and rigidity of the cold paste body respectively.     

Physicochemical characterisation of enzymatically hydrolysed derivatives of acetylated starch

Potato starch modified to different degrees by substitution with acetyl groups was the subject of this study undertaken to determine the influence of conditions of enzymatic hydrolysis on the surface-active properties of hydrolysates of acetylated starch. The effect of acetylation of starch preparation on its susceptibility to enzymatic hydrolysis in the membrane reactor was also considered. All hydrolysates of acetylated starch samples investigated were found to bring a decrease in the surface/interfacial tension, both at the air/water and the toluene/water interfaces. For binary hydrolysate-surfactant systems, the surface mole fractions in the mixed adsorbed monolayer at the air/water interface were estimated. For mixed systems, the synergism in reducing the surface tension at the air/water interface was observed. The experimentally obtained dynamic surface tension data for the aqueous solution of acetylated starch hydrolysates were used to estimate the diffusion coefficients. Particle size distributions of the hydrolysates formed in the aqueous solutions were compared to those of commercial maltodextrin.     

Starches from different botanical sources I: Contribution of amylopectin fine structure to thermal properties and enzyme digestibility

Fifteen starches from different botanical sources were selected to study the influence of structural features on thermal properties and enzyme digestibility. Morphological appearance, X-ray diffraction pattern, apparent amylose content, unit-chain length distribution of amylopectin, thermal properties and enzyme digestibility of starch varied with botanical source. It was demonstrated that the distribution of unit-chains of amylopectin significantly correlated with functional properties of the starches. Gelatinization temperature of native and retrograded starches decreased and increased with a relative abundance of unit-chains with an approximate degree of polymerization (DP) of 8–12 and 16–26, respectively (P<0.01). Similar unit-chain lengths also affected the enzyme digestibility of starch granules (P<0.01).     

Different isoforms of starch-synthesizing enzymes controlling amylose and amylopectin content in rice (Oryza sativa L.)

Starch, composed of amylose and amylopectin, greatly influences rice cooking and textural quality, which in turn is controlled by various isoforms of several enzymes. Activity of one or more isoforms of starch‐synthesizing enzymes results in various forms of starch structure based on the amylopectin chain length and average external, internal and core chain length distribution and hence results in varying physicochemical and cooking quality. Since the synthesis of starch is highly complex, it is crucial but essential to understand its biosynthetic pathway, starch structure and effects on the physicochemical properties that control eating and cooking quality, and alongside conduct research on gene/QTL mapping for use in marker-assisted selection (MAS) with a view to improve and select cultivars with most desirable range and class of rice starch properties. This article presents the updates on current understanding of the coordination among various enzymes/isoforms towards rice starch synthesis in endosperm and their effect on rice grain physicochemical, cooking and eating qualities. The efforts in identifying regions responsible for these enzymes by mapping the gene/QTLs have provided a glimpse on their association with physicochemical and cooking properties of rice and, hence, improvement is possible by modifying the allelic pattern, resulting in down or nil regulation of a particular enzyme. The clear understanding of the tissue specific coordination between enzyme isoforms and their subsequent effect in controlling eating and cooking properties will enhance the chances to manipulate them for getting desired range of amylose content (AC) and gelatinization temperature (GT) in improved cultivars through combining desired alleles through MAS.     

Characterization of puroindolines in the control of endosperm texture in common wheat lines with substitutions of homeologous group-5 chromosomes

The genetic control of grain hardness and its association with the specific friabilin content on starch granules of common wheat cultivars and lines with intervarietal substitutions of homeologous group-5 chromosomes were studied. A significant correlation was revealed between the technological parameters of grain hardness (mean size of flour particles) and the specific content of puroindolines on the starch surface estimated in terms of starch doses. The results obtained allowed the method of starch doses to be used to identify soft and hard wheat cultivars and lines based on an analysis of a single grain. The biochemical analysis confirmed the previously obtained estimates of flour-grinding properties of wheat cultivars and substitution lines and allowed specific genotypes to be characterized according to the composition of puroindolines. The influence of chromosomes 5D and 5A of donor wheat cultivars on the activity of the Ha loci of recipient cultivars was revealed and found to be associated with the composition of PIN products and with the expression of the Pina-D1 and Pinb-D1 genes.     

Expression of an amylosucrase gene in potato results in larger starch granules with novel properties

Main conclusion

Expression of amylosucrase in potato resulted in larger starch granules with rough surfaces and novel physico-chemical properties, including improved freeze–thaw stability, higher end viscosity, and better enzymatic digestibility. Starch is a very important carbohydrate in many food and non-food applications. In planta modification of starch by genetic engineering has significant economic and environmental benefits as it makes the chemical or physical post-harvest modification obsolete. An amylosucrase from Neisseria polysaccharea fused to a starch-binding domain (SBD) was introduced in two potato genetic backgrounds to synthesize starch granules with altered composition, and thereby to broaden starch applications. Expression of SBD–amylosucrase fusion protein in the amylose-containing potato resulted in starch granules with a rough surface, a twofold increase in median granule size, and altered physico-chemical properties including improved freeze–thaw stability, higher end viscosity, and better enzymatic digestibility. These effects are possibly a result of the physical interaction between amylosucrase and starch granules. The modified larger starches not only have great benefit to the potato starch industry by reducing losses during starch isolation, but also have an advantage in many food applications such as frozen food due to its extremely high freeze–thaw stability.     

Legume-Fortified Pasta. Impact of Drying and Precooking Treatments on Pasta Structure and Inherent In Vitro Starch Digestibility

The low glycaemic index of pasta can be attributed to its specific structure. A change in pasta structure can therefore lead to a change in its starch digestibility. The use of drastic drying conditions or the addition of non-traditional ingredients to durum wheat pasta was already demonstrated to affect its structure, leading to a modification of its starch digestibility. However, the combining effect of using different technological treatments and different raw materials on pasta structure and consequences on its starch digestibility pasta are still unknown. The objective of this work was therefore to determine the impact of different technological treatments on the structure and the in vitro starch digestibility (i.e. rapidly available glucose value) of legume-fortified pasta. Legume-fortified pasta was prepared from 65% of durum wheat semolina and 35% of split pea or faba bean flour. Four different technological treatments were applied: drying at low temperature of 55 °C (LT), drying with the application of a very high temperature of 90 °C at low moisture content (VHT.LM), lyophilisation, and precooking followed by LT drying. Legume-fortified pasta dried at LT served as a reference. Lyophilisation induced a higher starch digestibility that could be attributed to the high porosity of pasta and the weakness of its protein network. In contrast, VHT.LM drying and precooking treatment led to a lower in vitro starch digestibility, probably as a result of the strengthening of the protein network at a macromolecular level, protecting starch from enzymatic attack.     

Analysis of particle size reduction on overall surface area and enzymatic hydrolysis yield of corn stover

Particle size of lignocellulose materials is an important factor for enzymatic hydrolysis efficiency. In this study, corn stover was milled and sieved into different size fractions from 1.42, 0.69, 0.34, to 0.21 mm, and the corresponding enzymatic hydrolysis yields were 24.69, 23.96, 25.34, and 26.97 %, respectively. The results indicate that the hydrolysis yield is approximately constant with changing corn stover particle sizes in the experimental range. The overall surface area and the inner pore size measurement show that the overall specific surface area was less than 2 % with the half reduction of particle size due to the greater inner pore surface area. The scanning electron microscope photographs gave direct evidence of the much greater inner pore surface area of corn stover particles. This result provided a reference when a proper size reduction of lignocellulose materials is considered in biorefining operations.     

Endosperm enrichment and physicochemical properties of superior and inferior grain starch in super hybrid rice

• A significant asynchronous phenomenon exists in super hybrid rice because of the differences in spike and spikelet positions, which affect the accumulation and properties of starch. However, little is known about the endosperm enrichment and physicochemical properties of starch in superior and inferior grains in super hybrid rice.
• Rice YY2640 was selected as study material to investigate the enrichment and physicochemical properties of starch in superior and inferior grains in super rice using semi‐thin sections, X‐ray diffraction and related technologies.
• Superior grain filling was a continuous process, whereas inferior grain only started 8–10 days after anthesis. The order of starch accumulation starts in the central endosperm, then in the endosperm of the proximal vascular bundle and finally in the aleurone layer. Compared with the inferior grains, the superior grains have a higher 1000‐grain weight, apparent amylose content, total starch content, average starch granule size, relative crystallinity, solubility and a resonance peak ratio at 1022/995 cm^?1, whereas the swelling power and ratio of the resonance peak at 1045/1022 cm^?1 were lower. The final degree of hydrolysis of HCl, AAG and PPA of the superior grains were significantly lower than those of the inferior grains.
• The findings indicate that the different physicochemical properties of starch were mainly related to the development order of superior and inferior grains and the spatial enrichment of starch.