Functional properties of pregelatinized and cross-linked cassava starch obtained by extrusion with sodium trimetaphosphate

Cassava starch was cross-linked with sodium trimetaphosphate (STMP) on a Cerealtec single-screw extruder at different extrusion temperatures and concentrations of STMP and NaOH. The effect of variables on functional properties of the products was analysed by the response–surface methodology. The degree of substitution (DS) was influenced by NaOH and phosphorus level, and increased when their concentration increased. Extrusion temperature affected water absorption, cold viscosity and gel characteristics. The introduction of phosphate groups by cross-linking, with maximum DS of 1.5×10⁻⁴, increased the gel strength, water absorption index, resistance to high temperature and shear, and decreased gel cohesiveness, starch clarity and water solubility index. The products had different DS and degree of gelatinization and thus can be applied in several kinds of foods.     

Influence of Low Acyl and High Acyl Gellan Gums on Pasting and Rheological Properties of Rice Starch Gel

The pasting, viscoelastic, morphological and retrogradation properties of rice starch as affected by low acyl (LA) and high acyl (HA) gellan gums were studied. The additions of both LA and HA gums increased the peak and trough viscosities, while decreased the final and setback viscosities of rice starch paste. The starch-HA mixed pastes exhibited superior viscoelastic properties to the starch-LA pastes as evidenced by their higher storage modulus and lower loss tangent values. The starch-HA system exhibited higher resistance to the stress and more pronounced recovery rate in in-shear structural recovery test. The creep recovery data were well fitted by a 4-element Burger’s model. The shrinkage measurements showed that the addition of both hydrocolloids, especially the HA gellan gum retarded the retrogradation of rice starch gel during cold storage. It was concluded that the addition of LA and HA gellan gums modified the rheology and textural properties of rice starch gel in different ways and interacted under different mechanisms based on their molecular structures.     

Production and characteristics of inhibitory factor of raw starch digestion from Aspergillus niger

Summary The glucoamylase preparation of Aspergillus niger 19 inhibited the raw starch digestion by it at high enzyme concentration. The inhibitory factor (IF) was isolated from the glucoamylase preparation by heat treatment and purified by DEAE-Sephadex A-25 column chromatography, an initial Sephadex G-50 gel filtration followed by SP-Sephadex C-25 column chromatography (twice) and then second Sephadex G-50 gel filtration. The IF thus purified was homogenous in polyacrylamide gel electrophories. The inhibitory activity of IF increased with the increasing IF concentration but decreased with an increasing quantity of raw starch or enzyme concentration. The IF had no effect on the hydrolysis of boiled soluble starch. It was completely adsorbed onto raw starch. The IF had a molecular weight of about 10,500. It was abundant in hydroxy amino acids such as threonine and serine. Xylose, mannose, glucose, galactose, and galacturonic acid were present in it.     

Pullulanase from rice endosperm

Pullulanase (EC 3.2.1.41) in non-germinating seeds was compared with that in germinating seeds. Moreover, pullulanase from the endosperm of rice (Oryza sativa L., cv. Hinohikari) seeds was isolated and its properties investigated. The pI value of pullulanase from seeds after 8 days of germination was almost equal to that from non-germinating seeds, which shows that these two enzymes are the same protein. Therefore, the same pullulanase may play roles in both starch synthesis during ripening and starch degradation during germination in rice seeds. The enzyme was isolated by a procedure that included ammonium sulfate fractionation, DEAE-cellulofine column chromatography, preparative isoelectric focusing, and preparative disc gel electrophoresis. The enzyme was homogeneous by SDS/PAGE. The molecular weight of the enzyme was estimated to be 100 000 based on its mobility on SDS/PAGE and 105 000 based on gel filtration with TSKgel super SW 3000, which showed that it was composed of a single unit. The isoelectric point of the enzyme was 4.7. The enzyme was strongly inhibited by beta-cyclodextrin. The enzyme was not activated by thiol reagents such as dithiothreitol, 2-mercaptoethanol or glutathione. The enzyme most preferably hydrolyzed pullulan and liberated only maltotriose. The pullulan hydrolysis was strongly inhibited by the substrate at a concentration higher than 0.1%. The degree of inhibition increased with an increase in the concentration of pullulan. However, the enzyme hydrolyzed amylopectin, soluble starch and beta-limit dextrin more rapidly as their concentrations increased. The enzyme exhibited alpha-glucosyltransfer activity and produced an alpha-1,6-linked compound of two maltotriose molecules from pullulan.     

Effect of Multiple Freezing/Thawing Cycles on the Structural and Functional Properties of Waxy Rice Starch

The structural and functional properties of non-gelatinized waxy rice starch were investigated after 1, 3, 7, and 10 freezing/thawing cycles. Freezing caused an increasing damaged starch from 1.36% in native waxy rice starch to 5.77% in 10 freezing/thawing-treated starch (FTS), as evidenced by the cracking surface on starch granules. More dry matter concentration was leached, which was characterized by high amylopectin concentration (4.34 mg/mL). The leaching was accompanied by a decrease in relative crystallinity from 35.19% in native starch to 31.34% in 10 FTS. Freezing treatment also led to significant deviations in the functional characteristics, for instance decreased gelatinization temperature range, enthalpy, and pasting viscosities. The resistant starch content of 10FTS significantly decreased from 58.9% to 19%, whereas the slowly digested starch content greatly increased from 23.8% in native starch to 50.3%. The increase in susceptibility to enzyme hydrolysis may be attributed to porous granular surface, amylopectin leaching, and the decrease in the relative crystallinity caused by freezing water.     

Debranching enzyme concentration effected on physicochemical properties and α-amylase hydrolysis rate of resistant starch type III from amylose rice starch

The effect of debranching enzyme concentration on physicochemical properties and α-amylase hydrolysis rate of resistant starch type III from high amylose rice starch were studied. The pullulanase enzyme (8, 10, 12, 14 and 16 U/g starch) was introduced to modify amylopectin molecules of 15% (w/w) gelatinized rice starches at 55 °C for 16 h. The debranched starches with different degrees of hydrolysis (0.14–5.27%), and having 66.60–98.82% β-amylolysis limit were then induced at 4 °C for 16 h, afterward a one cycle of freeze–thaw process (?10/30 °C) was applied. The results showed that a pullulanase hydrolysis improved the degree of syneresis (51.64–54.85% from 8 to 16 U/g starch). Resistant starch content increased sharply as the amount of the enzyme increased, reaching the highest (19.81%) for a 12 U/g starch and decreased to 13.16% by 16 U/g starch. α-Amylase hydrolysis rate showed that incompletely-debranched had a lower estimated glycemic index than completely debranched rice starches. Microstructure of the selected RS III samples using X-ray diffraction and scanning electron microscopy revealed a crystal pattern change from A- to V-type pattern and formed a coarse honeycomb-like and a filamentous network structure.     

稻米品质性状对开放式空气二氧化碳浓度增高的响应

利用开放式空气CO2浓度增高(FACE)系统平台。研究大田栽培条件下粳稻武香粳14号稻米品质性状对CO2浓度增高200μmol·mol^-1的响应。结果表明．FACE处理稻谷的出糙率平均比CK高1．4个百分点,整精米率平均比CK低12．3个百分点,较低的供N水平有利于提高FACE条件下的出糙率．较高的供N水平有利于提高FACE条件下的整精米率;FACE处理的稻米垩白略有增加。垩白粒率平均比CK高11．9个百分点,垩白度平均比CK平均高2．8个百分点,较高的供N和供P水平有利于降低FACE条件下垩白大小、垩白粒率和垩白度;FACE处理稻米糊化温度平均比CK平均高0．52℃,胶稠度有提高的趋势,但对稻米直链淀粉含量影响较小,较高的供N和供P水平有利于降低FACE条件下稻米的直链淀粉含量,较低的供N和较高的供P水平有利于降低FACE条件下稻米胶稠度,较低的供N水平有利于降低FACE条件下稻米糊化温度;FACE处理使稻米蛋白质含量比CK平均低0．6个百分点,较低的供N和供P水平有利于降低FACE条件下稻米蛋白质含量。     

Impact of down-regulation of starch branching enzyme IIb in rice by artificial microRNA- and hairpin RNA-mediated RNA silencing

The inactivation of starch branching IIb (SBEIIb) in rice is traditionally associated with elevated apparent amylose content, increased peak gelatinization temperature, and a decreased proportion of short amylopectin branches. To elucidate further the structural and functional role of this enzyme, the phenotypic effects of down-regulating SBEIIb expression in rice endosperm were characterized by artificial microRNA (amiRNA) and hairpin RNA (hp-RNA) gene silencing. The results showed that RNA silencing of SBEIIb expression in rice grains did not affect the expression of other major isoforms of starch branching enzymes or starch synthases. Structural analyses of debranched starch showed that the doubling of apparent amylose content was not due to an increase in the relative proportion of amylose chains but instead was due to significantly elevated levels of long amylopectin and intermediate chains. Rices altered by the amiRNA technique produced a more extreme starch phenotype than those modified using the hp-RNA technique, with a greater increase in the proportion of long amylopectin and intermediate chains. The more pronounced starch structural modifications produced in the amiRNA lines led to more severe alterations in starch granule morphology and crystallinity as well as digestibility of freshly cooked grains. The potential role of attenuating SBEIIb expression in generating starch with elevated levels of resistant starch and lower glycaemic index is discussed.     

Development of reduced-fat mayonnaise using 4αGTase-modified rice starch and xanthan gum

In this study a disproportionating enzyme, 4-α-glucanotransferase (4αGTase), was used to modify the structural properties of rice starch to produce a suitable fat substitute in reduced-fat (RF) mayonnaise. The mayonnaise fat was partially substituted with the 4αGTase-treated starch paste at levels up to 50% in combination with xanthan gum and the physical and rheological properties of the modified RF mayonnaise samples were investigated. All mayonnaises prepared in this study exhibited shear thinning behavior and yield stress. Viscoelastic properties of mayonnaise were characterized using dynamic oscillatory shear test and it was observed that mayonnaises exhibited weak gel-like properties. The magnitude of elastic and loss moduli was also affected by 4αGTase-treated starch concentration and presence of xanthan gum. In relation to microstructure, RF mayonnaise prepared with 3.8 or 5.6 wt% of 4αGTase-treated starch and xanthan gum showed smaller droplets. The use of 5.6 wt% of 4αGTase-treated starch and 0.1 wt% of xanthan gum produced a RF mayonnaise with similar rheological properties and appearances as FF mayonnaise with gum. This study demonstrated a high feasibility for using 4αGTase-treated rice starch as a viable fat replacer in mayonnaise.     

复合调控Wx与SBE3基因对水稻籽粒直链淀粉含量的影响

颗粒淀粉合成酶（GBSS）和淀粉分支酶3（SBE3）是淀粉合成过程中的两个关键酶,这两个酶主要由耽和SBE3两个基因分别控制,它们的表达量直接影响直链淀粉和支链淀粉的含量比例。为了探讨水稻淀粉关键酶基因耽过量与SBE3干涉复合表达对直链淀粉含量的影响,构建了Wx过量表达与SBE3干涉结合的多基因表达载体,并通过农杆菌介导的方法将其导入日本晴水稻中。经过PCR检测分析获得了65株转基因阳性植株,半定量RT—PCR检测表明转基因株系中Wx基因表达量明显增加,而SBE3基因表达量显著减少。转基因株系籽粒透明度明显降低,直链淀粉含量比野生型的平均高45％,但是千粒重变化不大,与野生型相当。遗传分析表明这些转基因株系多数可稳定遗传。     

Effect of gibberellic acid on starch degrading enzymes in leaves of Digitaria decumbens

Tropical ‘Pangola’ digitgrass was treated with gibberellic acid (GA) and subjected to 30° or 10° nights. The starch degrading enzymes of the leaves were separated by polyacrylamide gel electrophoresis. Densitometer tracings of gels negatively stained by starch-iodine showed the presence of seven bands regardless of treatment of plants. GA treatment increased starch degrading enzyme activity in plants subjected to 10° to the level of activity found in 30° untreated (control) plants and, additionally, enhanced enzyme activity in plants at 30°. GA treatment of 10° plants decreased sucrose and starch levels when compared to levels found in untreated 10° plants. The action of GA in reversing the effects of 10° nights on ‘Pangola’ leaves was found to be the result of a quantitative increase in activity of existing enzyme forms rather than production of isozymes.     

Enzymic Mechanism of Starch Breakdown in Germinating Rice Seeds: III. alpha-Amylase Isozymes

The formation of amylase isozymes in germinating rice (Oryza sativa) seeds was studied by isoelectric focusing on polyacrylamide gel disc electrophoresis. Time sequence comparisons of the amylase zymogram were made between extracts from gibberellic acid-treated embryoless and embryo-attached half-endosperm of rice seeds. In both cases, 4 major and 9 to 10 minor isozyme bands were detectable at the maximal stage of the enzyme induction. However, in the embryo-attached half-seeds, bands started to diminish after the 5th day of incubation, in agreement with the results of time sequence analyses of enzyme activities. Nearly identical patterns of amylase isozyme bands on a polyacrylamide gel disc electrophoresis in combination with isoelectric focusing indicate the intrinsic role of gibberellic acid in the starch breakdown in germinating rice seeds. We tentatively assign the newly synthesized enzymes to be α-amylases based on experimental results concerning the lability of the preparation on a prolonged treatment at pH 3.3 and the stability on heat treatment for 15 minutes at 70 C.     

淀粉分支酶3存在于水稻胚乳的淀粉粒之中

对水稻胚乳淀粉颗粒结合的淀粉分支酶进行了研究.酶活性分析表明水稻胚乳中存在着与淀粉颗粒结合的淀粉分支酶.氨基酸测序分析结果表明结合于水稻胚乳淀粉粒的淀粉分支酶是分子量为84 kD的淀粉分支酶3(rice starch branching enzyme 3; RBE3).从开花后5 d到种子成熟,淀粉颗粒结合的RBE3蛋白都保持较为稳定的含量.Northern 分析表明水稻胚乳发育过程中RBE4最先表达而RBE3和RBE1的表达滞后.综合以上研究结果说明RBE3存在于水稻胚乳的淀粉之中是由于RBE3与淀粉葡聚糖链具有较高亲和性而难以和葡聚糖链解离,进而随着淀粉粒的增长而被其包裹.     

Excess nickel modulates activities of carbohydrate metabolizing enzymes and induces accumulation of sugars by upregulating acid invertase and sucrose synthase in rice seedlings

The effects of increasing concentrations of nickel sulfate, NiSO₄ (200 and 400 μM) in the growth medium on the content of starch and sugars and activity levels of enzymes involved in starch and sugar metabolism were examined in seedlings of the two Indica rice cvs. Malviya-36 and Pant-12. During a 5–20 day growth period of seedlings in sand cultures, with Ni treatment, no definite pattern of alteration in starch level could be observed in the seedlings. In both roots and shoots of the seedlings Ni treatment led to a significant decrease in activities of starch degrading enzymes α-amylase, β-amylase, whereas starch phosphorylase activity increased. The contents of reducing, non-reducing, and total sugars increased in Ni-treated rice seedlings with a concomitant increase in the activities of sucrose degrading enzymes acid invertase and sucrose synthase. However, the activity of sucrose synthesizing enzyme sucrose phosphate synthase declined. These results suggest that Ni toxicity in rice seedlings causes marked perturbation in metabolism of carbohydrates leading to increased accumulation of soluble sugars. Such perturbation could serve as a limiting factor for growth of rice seedlings in Ni polluted environments and accumulating soluble sugars could serve as compatible solutes in the cells under Ni toxicity conditions.     

Structural and physicochemical properties of starch gels prepared from partially modified starches using Thermus aquaticus 4-α-glucanotransferase

The modified starch gels prepared from partial enzyme treatments (1, 3, and 6 U/g starch; 2-h incubation) of the corn and rice starch pastes using Thermus aquaticus 4-α-glucanotransferase (TAαGT) were investigated for their molecular characteristics, microstructures, and physicochemical properties. Unlike the native and partially modified normal starches, the native and partially modified waxy starches could not form gels strong enough for textural analysis after 24 h for gel setting. Features of the partially modified normal starches were the specific apparent amylose contents and maximum iodine absorption wavelength (λ_max, ∼567 nm), as well as the tri-modal molecular weight profiles and flatter side-chain distributions. Also, the partially modified normal starch gels possessed fractured surfaces with discontinuous crystalline fibrous assembly that differed from the native starch gels’ porous continuous network, which resulted in more brittle, rigid, and resilient gels compared with the native gels.     

自絮凝酵母SPSC01悬浮液的流变行为

用旋转黏度计测定了自絮凝颗粒酵母悬浮液的流变特性,并考察了其流变特性的影响因素,如菌体生物量、葡萄糖质量浓度、温度等。结果表明：自絮凝颗粒酵母悬浮液呈假塑性非牛顿流体,其流变特性服从幂律指数模型,随着菌体浓度的增大,稠度系数增大,流动行为指数减小;絮凝悬浮液的表观黏度随着糖浓度的增加有所增加,同一生物量下稠度系数k随着糖浓度的增加而增加,流动行为指数n随着糖浓度的增加变化很小,悬浮液的表观黏度随着温度的升高而降低;相同生物量下的流变指数随温度的升高而升高,而稠度系数随温度升高有所下降。     

Influence of Cross-linked Waxy Maize Starch on the Aggregation Behavior of Casein Micelles During Acid-induced Gelation

The influence of cross-linked waxy maize starch on the aggregation behavior of casein micelles was investigated using a combination of physico-chemical techniques. Milk was homogenized at two different temperatures (55 and 65 °C) and then heated at 95 °C for 5 min in a pilot scale system. The possible interactions between modified starch and milk proteins during lactic acid fermentation were evaluated. While 1% starch did not show differences in the whey protein complexes formed during heating compared to milk with no starch (as measured by size exclusion chromatography), a higher (2.5%) concentration of starch clearly showed an increased amount of heat-induced whey protein aggregates. The gelation pH also increased significantly with 2.5% starch compared to that of the control samples. The storage modulus (G′) increased with increasing levels of starch, and confocal microscopy confirmed that the microstructure of the casein gels was altered by the presence of modified starch. Milk-starch mixtures preheated and homogenized at 55 or 65 °C exhibited similar physico-chemical behavior during acidification. The results suggested a lack of interaction between starch granules and casein micelles during acidification, and scanning electron microscopy images collected with a self-assembled monolayer technique also confirmed that starch granules were not attached to milk caseins but only embedded in the protein gel matrix.     

Some properties of starch branching enzyme from indica rice endosperm (Oryza sativa L.)

Starch branching enzyme (α-1,4-glucan: α-1,4-glucan-6-glycosyl transferase; EC 2.4.1.18) catalyzes the formation of the α-1,6-bond in branched starch molecules such as amylopectin. Some characteristics of starch branching enzyme in rice endosperm (Oryza sativa L.) were determined because of the importance of starch structure for rice quality. Two or three peaks of starch branching enzyme activity were resolved by anion-exchange chromatography of extracts from high amylose rice. The properties of rice starch branching enzyme were similar to those found for the enzyme from other plant sources except for a much lower molecular weight. Rice branching enzyme had an apparent molecular weight of 40 000 as estimated by gel permeation chromatography. Multiple forms of starch branching enzyme could also be resolved in milled rice, suggesting that relationships between starch quality and characteristics of starch branching enzyme could be examined in the mature grain after harvest.     

Seed yield and plant biomass increases in rice are conferred by deregulation of endosperm ADP-glucose pyrophosphorylase

In this work we test the hypothesis that yield of rice ( Oryza sativa L.) can be enhanced by increasing endosperm activity of ADP-glucose pyrophosphorylase (AGP), a key enzyme in starch biosynthesis. The potential for increases in yield exist because rice initiates more seeds than are taken to maturity and possesses excess photosynthetic capacity that could be utilized if there were more demand for assimilate. Following an approach already shown to be successful in wheat, experiments were designed to increase demand for assimilate by increasing the capacity for starch synthesis in endosperm. This was accomplished by transforming rice with a modified maize AGP large subunit sequence ( Sh2r6hs) under control of an endosperm-specific promoter. This altered subunit confers upon AGP decreased sensitivity to allosteric inhibition by inorganic phosphate (Pi) and enhanced heat stability, potentially leading to higher AGP activity in vivo. The Sh2r6hs transgene increased AGP activity in developing endosperm by 2.7-fold in the presence of Pi. Increases in AGP activity in transgenic seeds compared with controls were maximal between 10-15 days after anthesis. Starch content of individual seeds at harvest was not increased, but seed weight per plant and total plant biomass were each increased by more than 20%. Increased endosperm AGP activity thus stimulates setting of additional seeds and overall plant growth rather than increasing yield of seeds already set. Results demonstrate that deregulation of endosperm AGP increases overall plant sink strength, leading to larger, more productive plants in a manner similar to that in wheat having similar genetic modification.