谷物籽粒淀粉研究进展

谷物籽粒是人类最基本的粮食,其胚乳淀粉与品质有密切的关系.本文就谷物胚乳淀粉的合成积累过程、谷物淀粉合成酶的作用与特性、淀粉特性与品质的关系以及基因工程在改良作物淀粉品质方面的研究进展进行了综述,并对谷物籽粒淀粉进一步的研究提出了展望,为谷物淀粉品质育种提供参考.     

Starch granule initiation in Arabidopsis thaliana chloroplasts

The initiation of starch granule formation and the mechanism controlling the number of granules per plastid have been some of the most elusive aspects of starch metabolism. This review covers the advances made in the study of these processes. The analyses presented herein depict a scenario in which starch synthase isoform 4 (SS4) provides the elongating activity necessary for the initiation of starch granule formation. However, this protein does not act alone; other polypeptides are required for the initiation of an appropriate number of starch granules per chloroplast. The functions of this group of polypeptides include providing suitable substrates (maltooligosaccharides) to SS4, the localization of the starch initiation machinery to the thylakoid membranes, and facilitating the correct folding of SS4. The number of starch granules per chloroplast is tightly regulated and depends on the developmental stage of the leaves and their metabolic status. Plastidial phosphorylase (PHS1) and other enzymes play an essential role in this process since they are necessary for the synthesis of the substrates used by the initiation machinery. The mechanism of starch granule formation initiation in Arabidopsis seems to be generalizable to other plants and also to the synthesis of long-term storage starch. The latter, however, shows specific features due to the presence of more isoforms, the absence of constantly recurring starch synthesis and degradation, and the metabolic characteristics of the storage sink organs.     

禾谷类作物淀粉合酶

淀粉合酶是禾谷类作物淀粉合成所必需的一类酶.根据淀粉合酶家族成员的氨基酸序列的相似性,分别介绍了一个颗粒性淀粉合酶亚家族和四个可溶淀粉合酶亚家族的组成、基因结构和表达特点,并从转录、转录后和翻译后水平上对这些基因的表达调控做了概述.     

Physicochemical properties of endosperm and pericarp starches during maize development

Endosperm starch and pericarp starch were isolated from maize (B73) kernels at different developmental stages. Starch granules, with small size (2–4 μm diameter), were first observed in the endosperm on 5 days after pollination (DAP). The size of endosperm-starch granules remained similar until 12DAP, but the number increased extensively. A substantial increase in granule size was observed from 14DAP (diameter 4–7 μm) to 30DAP (diameter10–23 μm). The size of starch granules on 30DAP is similar to that of the mature and dried endosperm-starch granules harvested on 45DAP. The starch content of the endosperm was little before 12DAP (less than 2%) and increased rapidly from 10.7% on 14DAP to 88.9% on 30DAP. The amylose content of the endosperm starch increased from 9.2% on 14DAP to 24.2% on 30DAP and 24.4% on 45DAP (mature and dried). The average amylopectin branch chain-length of the endosperm amylopectin increased from DP23.6 on 10DAP to DP26.9 on14DAP and then decreased to DP25.4 on 30DAP and DP24.9 on 45DAP. The onset gelatinization temperature of the endosperm starch increased from 61.3 °C on 8DAP to 69.0 °C on 14DAP and then decreased to 62.8 °C on 45DAP. The results indicated that the structure of endosperm starch was not synthesized consistently through the maturation of kernel. The pericarp starch, however, showed similar granule size, starch content, amylose content, amylopectin structure and thermal properties at different developmental stages of the kernel.     

Improved cold starch hydrolysis with urea addition and heat treatment at subgelatinization temperature

We explore how the presence of urea can influence the kinetics of amylolysis, with a long-term objective of developing practical and energy efficient bioconversion protocols. In this study, triticale and corn starches were hydrolyzed by a granular starch hydrolyzing enzyme with or without addition of urea and a pre-heating treatment at subgelatinization temperature. Differential scanning calorimetry showed that the gelatinization parameters of triticale and corn starches were negatively correlated with the urea concentration in the starch suspension. Addition of urea did not significantly affect starch amylolysis by the granular starch hydrolyzing enzyme at 30 °C. However when pre-heating at a higher yet sub-gelatinization temperature (50 °C for triticale and 61 °C for corn, 5 °C below the onset of starch gelatinization) for 30 min, the presence of urea greatly facilitated the amylolysis of both tricticale and corn starches. Scanning electron microcopy revealed starch granule mophological changes to a porous structure in residual starch granules/fragments rich in resistant starch. This means that the amylolysis pattern in the presence of urea was fundamentally changed, and urea disrupts starch hydrogen bonds effectively with heating treatment at a sub-gelatinization temperature. This treatment combination increased both starch hydrolysis rate and extent. Since extra energy was not necessary to gelatinize starch, this method may benefit starch and bio-enthanol industries to reduce the costs of starch hydrolysis.     

Effects of protein on crosslinking of normal maize, waxy maize, and potato starches

Channels of maize starch granules are lined with proteins and phospholipids. Therefore, when they are treated with reagents that react at or near the surfaces of channels, three types of crosslinks could be produced: protein–protein, protein–starch, starch–starch. To determine which of these may be occurring and the effect(s) of channel proteins (and their removal) on crosslinking, normal and waxy maize starches were treated with a proteinase (thermolysin, which is known to remove protein from channels) before and after crosslinking, and the properties of the products were compared to those of a control (crosslinking without proteinase treatment). After establishing that treatment of starch with thermolysin alone had no effect on the RVA trace, three reaction sequences were used: crosslinking alone (CL), proteinase treatment before crosslinking (Enz-CL), proteinase treatment after crosslinking (CL-Enz). Two crosslinking reagents were used: phosphoryl chloride (POCl₃), which is known to react at or near channel surfaces; STMP, which is believed to react throughout the granule matrix. Three concentrations of POCl₃ (based on the weight of starch) were used. For both normal maize starch (NMS) and waxy maize starch (WMS) reacted with POCl₃, the trends were generally the same, with apparent relative degrees of crosslinking indicated to be CL-Enz = CL > Enz-CL, but the effects were greater with NMS and there were differences when different concentrations of reagent were used. The basic trends were the same when potato starch was used in the same experiments. Crosslinking with STMP was done both in the presence and the absence of sodium sulfate (SS). Both with and without SS and with both NMS and WMS, the order of indicated crosslinking was generally the same as found after reaction with POCl₃, with the indicated swelling inhibition being greater when SS was present in the reaction mixture. Examination of the maize starches with a protein stain indicated that channel protein was removed by treatment with thermolysin when the proteinase treatment occurred before crosslinking with either POCl₃ or STMP, but only incompletely or not at all if the treatment with the proteinase occurred after crosslinking. Because the crosslinking reactions were less effective when the protein was removed, the results are tentatively interpreted as indicating that they involved protein molecules, although there may not be a direct relationship.     

Potato powders prepared by successive cooking-process depending on resistant starch content affect the intestinal fermentation in rats

The effects of resistant starch (RS) in dry potato powders prepared by various processes on intestinal fermentation in rats were assessed. Rats were fed raw potato powder (RP), blanched potato powder (BP), steamed potato powder (SP), or drum-dried potato powder (DP) for 4 weeks. The cecal RS content was significantly higher in the RP group than in the control diet (CN) group and other dry potato powder groups. Cecum pH was significantly lower in the RP group compared to the CN group, and was also significantly lower than that in the SP, BP, and DP groups. Lactic acid bacteria levels in the RP group were significantly higher than those in the CN group, and levels in the SP group also increased relative to the control group. Lactobacillus levels in the RP group were higher than in the CN and other dry potato powder groups. Cecal short-chain fatty acid (SCFA) concentrations in the RP group followed by the SP group exhibited significantly higher levels relative to the control levels. Dry potato powders containing RS produced during the cooking process may represent a useful food material that increases intestinal concentrations of SCFA and enhances the growth of certain lactic acid bacteria.     

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.     

A genetic strategy generating wheat with very high amylose content

Resistant starch (RS), a type of dietary fibre, plays an important role in human health; however, the content of RS in most modern processed starchy foods is low. Cereal starch, when structurally manipulated through a modified starch biosynthetic pathway to greatly increase the amylose content, could be an important food source of RS. Transgenic studies have previously revealed the requirement of simultaneous down‐regulation of two starch branching enzyme (SBE) II isoforms both located on the long arm of chromosome 2, namely SBEIIa and SBEIIb, to elevate the amylose content in wheat from ~25% to ~75%. The current study revealed close proximity of genes encoding SBEIIa and SBEIIb isoforms in wheat with a genetic distance of 0.5 cM on chromosome 2B. A series of deletion and single nucleotide polymorphism (SNP) loss of function alleles in SBEIIa, SBEIIb or both was isolated from two different wheat populations. A breeding strategy to combine deletions and SNPs generated wheat genotypes with altered expression levels of SBEIIa and SBEIIb, elevating the amylose content to an unprecedented ~85%, with a marked concomitant increase in RS content. Biochemical assays were used to confirm the complete absence in the grain of expression of SBEIIa from all three genomes in combination with the absence of SBEIIb from one of the genomes.     

RVA analysis of mixtures of wheat flour and potato, sweet potato, yam, and cassava starches

Rapid visco analysis (RVA) was performed to study the pasting properties of mixtures of wheat flour and tuber starches, i.e., potato starch (PS), sweet potato starch (SPS), yam starch (YS), and cassava starch (CS), at 10–50% starch in the mixtures. Lower phosphorus and higher amylose contents were observed in CS, followed by YS, SPS, and PS. The peak, breakdown, final, and setback viscosities of the control wheat flour were lower than those of the control PS, SPS, YS, and CS. The peak viscosity of wheat–PS mixtures was higher than those of the wheat–SPS, wheat–YS, and wheat–CS because of the higher phosphorus and lower amylose content of PS, which resulted in higher swelling of PS than that of SPS, YS, and CS. The breakdown viscosities increased as the starch content of the PS, SPS, and CS in the mixtures increased to up to the 50%, and the values tended to decrease in the wheat–YS mixture. The setback viscosities of wheat–SPS, wheat–YS, and wheat–CS increased significantly as the starch content increased from 10% to 50%, and that of wheat–PS dropped dramatically at 50%. The findings in this work provide evidence that tuber starches could be used as a partial substitute for wheat flour in some wheat-based products.     

羧甲基淀粉崩解性能研究

本研究考察了崩解剂羧甲基淀粉（ＣＭＳ）的取代度（ＤＳ）和ＣＭＳ中Ｈ／Ｎａ（游离羧基和成盐羧基比值）改变对其吸水溶胀性能和崩解性能的影响。并应用红外光谱,Ｘ－射线衍射和扫描电镜等方法探求ＣＭＳ的结构与性能的关系。     

Detection of the plasma volume expander hydroxyethyl starch in human urine

The plasma volume expander hydroxyethyl starch (HES) is usually administered in cases of hypovolaemic shocks but in 1998 the press reported its misuse in endurance sports. Since January 2000, it has been put on the list of prohibited substances of the International Olympic Committee (IOC) and its misuse is to ban by doping controls. Therefore, a rapid method enabling the screening for HES in human urine was developed which can be easily adopted by IOC laboratories to analyse routine urine samples for this remedy. Excretion study urine samples obtained from patients treated with HES, blank urine specimen and reference standards, were hydrolysed with hydrochloric acid and without any further purification of the resulting monosaccharides their per-timethylsilylated derivatives were performed. By means of gas chromatography–mass spectrometry the products were separated and the α- and β-isomers of glucose, 2-, 3- and 6-hydroxyethyl glucose derivatives were identified. Typical ion traces of 2- and 3-substituted glucose (m/z 248, m/z 261 and m/z 235, m/z 248, respectively) support the fast determination of the substances whose electron impact mass spectra are presented and discussed.     

The phenotype of soluble starch synthase IV defective mutants of Arabidopsis thaliana suggests a novel function of elongation enzymes in the control of starch granule formation

All plants and green algae synthesize starch through the action of the same five classes of elongation enzymes: the starch synthases. Arabidopsis mutants defective for the synthesis of the soluble starch synthase IV (SSIV) type of elongation enzyme have now been characterized. The mutant plants displayed a severe growth defect but nonetheless accumulated near to normal levels of polysaccharide storage. Detailed structural analysis has failed to yield any change in starch granule structure. However, the number of granules per plastid has dramatically decreased leading to a large increase in their size. These results, which distinguish the SSIV mutants from all other mutants reported to date, suggest a specific function of this enzyme class in the control of granule numbers. We speculate therefore that SSIV could be selectively involved in the priming of starch granule formation.     

Influence of incubation temperature and time on resistant starch type III formation from autoclaved and acid-hydrolysed cassava starch

Raw cassava starch, having 74.94 and 0.44 g/100 g resistant starch type II and III (RS II and RS III), respectively, was autoclaved at 121 °C in water, 1, 10 or 100 mmol/L lactic acid. The formation of RS III was evaluated in relation to variable incubation temperature (−20 to 100 °C), incubation time (6–48 h) and autoclaving time (15–90 min). Negligible to low quantities of RS III (0.59–2.42 g/100 g) were formed from autoclaved starch suspended in 100 mmol/L lactic acid, whereas intermediate to high quantities (2.68–9.97 g/100 g) were formed from autoclaved starch suspended in water, 1 or 10 mmol/L lactic acid, except for treatments with water or 10 mmol/L lactic acid incubated at 100 °C for 6 h (1.74 g/100 g). Autoclaving times corresponding to maximum RS III contents were 15 and 45 min for water and 10 mmol/L lactic acid, respectively. Whereas, the RS III fractions from cassava starch suspended in water had melt transitions between 158 and 175 °C with low endothermic enthalpies (0.2–1.6 J/g), the thermal transitions of the acid treated samples were indistinct.     

Effects of the activities of key enzymes involved in starch biosynthesis on the fine structure of amylopectin in developing rice (Oryza sativa L.) endosperms

The dynamic changes of the activities of enzymes involving in starch biosynthesis, including ADP-glucose pyrophosphorylase (AGPase), soluble starch synthases (SSS), starch branching enzyme (SBE) and starch debranching enzymes (DBE) were studied, and changes of fine structure of amy- lopectin were characterized by isoamylase treatment during rice grain development, using trans anti-waxy gene rice plants. The relationships between the activities of those key enzymes were also analyzed. The amylose synthesis was significantly inhibited in transgenic Wanjing 9522, but the total starch content and final grain weight were less affected as compared with those of non-transgenic Wanjing 9522 rice cultivar. Analyses on the changes of activities of enzymes involving in starch bio- synthesis showed that different enzyme activities were expressed differently during rice endosperm development. Soluble starch synthase is relatively highly expressed in earlier stage of endosperm de- velopment, whilst maximal expression of granule-bound starch synthase (GBSS) occurred in mid-stage of endosperm development. No obvious differences in changes of the activities of AGPase and SBE between two rice cultivars investigated, except the DBEs. Distribution patterns of branches of amy- lopectin changed continually during the development of rice grains and varied between two rice culti- vars. It was suggested that amylopectin synthesis be prior to the synthesis of amylose and different enzymes have different roles in controlling syntheses of branches of amylopectin.     

Resistant glutarate starch from adlay: Preparation and properties

Reaction conditions were optimized to increase the content of resistant starch in adlay starch using esterification with glutaric acid, and the physicochemical properties of the prepared glutarate starches were investigated. Different amounts of glutaric acid (0.1–0.5 g/g starch, dry weight basis) were reacted with adlay starch at various temperatures (70–130 °C) and reaction times (3–9 h). The resistant starch levels increased with increased glutaric acid content, reaction temperature, and reaction time. The color difference was mainly affected by reaction time. The highest resistant starch content (RS 66%) was obtained using conditions of 0.4 g glutaric acid/g starch, 115 °C, and 7.5 h, with a color difference of 10.24. After digestion with α-amylase and amyloglucosidase, the water-soluble fraction of glutarate starch had more oligosaccharides than high-amylose maize starch (RS 43%). FT-IR and solid-state NMR detected carbonyl groups in the glutarate starch, indicating the formation of cross-linkages through esterification. The granular structure of the glutarate starches was not destroyed and they retained birefringence. After heating with an excess of water, the granules kept their shape but lost their birefringence. The glutarate starches had low solubility in both cold and hot water, and the resistant starch contents were unchanged after heating due to the restriction of swelling by cross-linking. The glutarate starches had a similar chain-length distribution to raw starch, indicating that acid hydrolysis took place at branching points in the amorphous region. Furthermore, the glutarate starches possessed a weaker crystalline region, more diverse double helical chains, and lower enthalpy than raw starch.     

瓦楞纸板用淀粉粘合剂制造方法的探讨

介绍了瓦楞纸板用淀粉粘合剂的制备方法,包括过氧化氢、次氯酸钠和高锰酸钾等氧化剂的氧化法;并对淀粉粘合剂质量的影响因素进行了讨论。这将有助于淀粉粘合剂的生产和应用。     

马铃薯表观淀粉含量与直链淀粉含量相关性研究

测定了48个不同马铃薯品种表观淀粉含量以及块茎中和淀粉中直链淀粉含量,对表观淀粉含量和块茎中直链淀粉含量间,表观淀粉含量和淀粉粒直链淀粉含量间进行了相关分析。结果表明：表观淀粉含量和块茎中直链淀粉含量间相关显著,表观淀粉含量和淀粉粒直链淀粉含量间相关不显著,且中熟和晚熟基因型表观淀粉含量和淀粉粒直链淀粉含量间相关也不显著,这些结论将为淀粉生物合成的理论研究和淀粉品质改良提供基本的表型数据。     

An improved model for analyzing the small angle x-ray scattering of starch granules

The structure of starch was studied using small-angle x-ray scattering (SAXS). The scattering data was modeled by considering a finite stack of alternating lamellae that are allowed to fluctuate both along the layer repeat direction and along the transverse layer direction. Analysis in this way of the SAXS data from starch allowed fresh insights into the native structure of several starch species, particularly potato starch. The novel model presented in this work was able to capture the experimentally observed SAXS patterns much better than previous models, which did not incorporate transverse layer fluctuations.