不同提取方法对面包果淀粉性质的影响

本文采用湿磨法、碱法、中性蛋白酶法提取面包果淀粉,研究三种不同提取方法对面包果淀粉性质的影响。结果表明:湿磨法、碱法、中性蛋白酶法的提取率分别为23.90%、66.25%、66.05%,而碱法、中性蛋白酶法无显著性差异(P>0.05);面包果淀粉提取破损情况:碱法(4.20%)>中性蛋白酶法(2.28%)>湿磨法(1.92%);湿磨法提取面包果淀粉中蛋白质和脂肪残留量最多,碱法和中性蛋白酶法的残留量相差较小;不同方法提取的面包果淀粉溶解度和膨胀度均随温度的升高而升高,湿磨法提取的面包果淀粉溶解度和膨胀度均最大,碱法和中性蛋白酶法相差较小。经综合评价得出,中性蛋白酶法为面包果淀粉的最佳提取方法。本研究为面包果淀粉提供了一种新型绿色提取工艺,以期为面包果淀粉获得更加经济环保的工业化生产提供理论依据。     

Polarization Colors of Lightly Iodine‐stained Maize Starches for Amylose‐Extender and Related Genotypes in the W64A Inbred Line

Previous qualitative research showed that for some maize endosperm genotype starches the color of variably iodine‐stained starch granules observed by bright field microscopy (BFM) was different from the color of identically stained granules observed by polarized light microscopy (PLM). One objective of the present study was to determine the polarization color for a variety of high‐amylose and other starch genotypes in an identical genetic background. A secondary objective was to determine the iodine concentration dependence of polarization colors for the samples. Starches from the W64A inbred line were obtained from the following genotypes: normal, wx, ae, du, su2, ae wx, ae du, ae su2, and du su2. Starches were stained with iodine solutions ranging from 0.02 to 0.075% and viewed with BFM and PLM, using an auto‐exposure digital camera function. Most starches showed the first appreciable color at about 0.04percnt; I₂. Unlike normal and non‐ae‐containing starches, ae starch showed a pink polarization color, despite its blue color in bright field. Heterogeneity in polarization color was observed both within and among granules. Double mutant starches containing ae showed variable effects, depending on the combination. It is suggested that the pink polarization color of ae starch may be due to a lack of symmetrical orientation of iodinecomplexed amylose in these granules     

Evaluation of starch analysis methods for feed samples

The objectives of this work were to evaluate the efficacies of commercial starch analyses and of starch analysis extraction and gelatinisation procedures. In Study 1, accuracy and specificity of commercially available starch analyses were evaluated with six co‐operating laboratories (five commercial, one university). Results from 11 test samples showed three laboratories with recoveries of purified starch of 92 g kg⁻¹ or less. Three and four laboratories had inflated values when samples contained glucose or sucrose, respectively. Analyses appeared to have good specificity for glucose. Incompleteness of starch detection and interference by non‐starch carbohydrates can affect commercially available analyses. In Study 2, extraction with 80:20 ethanol/water (v/v; 80EtOH) or 90:10 ethanol/water (v/v; 90EtOH) to remove low‐molecular‐weight carbohydrates, and gelatinisation with heat, alkali (KOH), 6 M urea or 8 M urea were evaluated. Extraction with 80EtOH or 90EtOH reduced interference from non‐starch carbohydrates. Gelatinisation with heat was adequate for good recoveries of starch glucose for both control (non‐extracted) and 80EtOH‐extracted samples; gelatinisation with alkali was required for 90EtOH‐extracted samples. Recoveries of pure starch samples were greatest with no extraction and heat gelatinisation. 80EtOH extraction with heat gelatinisation appears to be an adequate preparation method when removal of low‐molecular‐weight carbohydrates is desired. © 2000 Society of Chemical Industry     

Role of the Recessive Amylose-extender Allele in Starch Biosynthesis of Maize

To investigate the role of the recessive amylose extender (ae) allele in starch biosynthesis, four endosperm groups with different doses of ae were harvested at 10, 14, 18, 22, 28 and 36 days after pollination, analyzed by disc-gel electrophoresis. After incubation with ADP-glucose and amylose primer multiple forms of starch synthetases were detected. Of the seven major bands, the four slow moving bands were observed only in the presence of ae allele and at limited stages of endosperm development. Appearance of these four bands coincided with the rapid increase of amylose content in the starch. The presence of modified amylopectin synthesized by the enzymes, which were affected by the ae allele, is suggested.     

In situ Raman microscopy of starch granule structures in wild type and ae mutant maize kernels

A method developed for in situ imaging of starch granule structure in dry seeds has been applied to compare the starch granule structures found in wild type and ae mutant maize kernels. In the isogenic ae mutant the activity of the starch branching enzyme IIb is inhibited, which gives rise to a high amylose starch. The granule structures in the wild type samples have been found to be homogeneous, whereas those in the ae mutant are grossly heterogeneous within individual granules, between granules within individual cells, and between cells across the endosperm. The level of heterogeneity observed in situ appears to be more marked than that previously reported for studies on isolated ae mutant starches. Iodine/potassium iodide staining and polarised light microscopy have been used together with Raman microscopy, which has allowed high‐resolution mapping of the composition and physical state of the structures within the granules, to probe the origins of the heterogeneity of the starch structures. Although the mutation inhibits the activity of the branching enzyme within the granules, and both the composition and level of crystallinity within and between granules is variable, the major origin of the heterogeneity of the granule architecture appears to result from significant changes in the assembly and packaging of the crystalline structures within the granule. It is suggested that this arises due to the mutation of the starch branching enzyme introducing defects into the self‐assembly of the crystalline structure, resulting in an accumulation of defects and increased randomisation of the granule structure.     

The Influences of Chain Length of Amylopectin on Resistant Starch in Rice (Oryza sativa L.)

Amylopectin is the principle component of starch. To elucidate the relationships between amylopectin and resistant starch content, six rice mutants with altered fine structure of amylopectin were selected for comparative studies with the primary wild type and two types of amylose‐extender (ae) mutants. Significant differences in resistant starch content were observed among mutants with similarity or differences in amylose levels. Mutants high in resistant starch had significantly increased proportions of short amylopectin chains with DP≤12, decreased levels of intermediate chains with size of 13≤DP≤36, and decreased fractions of long chains with DP≥37. Additionally, there was a mutant different to ae, which was characterized by an increased level of short chains with 8≤DP≤12 and 13≤DP≤24, and a decreased proportion of long chains with DP≥37. The increased contents of short chains with 8≤DP≤12 and decreased of intermediate and long chains with 24≤DP were clearly associated with the increase of resistant starch in rice.     

Structural and Physicochemical Properties of Endosperm Starches Possessing Different Alleles at the Amylose-extender and Waxy Locus in Maize (Zea mays L.)

The structure and some physicochemical properties of endosperm starches from seven amylose-extender (ae) and two waxy (wx) alleles of maize (Zea mays L.) were studied. Starches prepared from mature kernels of six ae mutants, Oh43 inbred line ae (standard ae), ae-RWB-2 and ae-RWB-3, and W23 × L317 hybrid line ae-PP, ae-Bol 561 and ae-emll, were uniquely ae type, as was concluded from B type X-ray diffractograms; high gelatinization temperatures determined by differential scanning calorimetry (DSC); with poor starch-granule digestibility to amylase; high amylose (37—45 %) and high intermediate fraction (13—18 %) contents; and low ratios (1.0—1.2) of long α-1,4-chains to short α-1,4-chains of amylopectin determined by gel permeation chromatography (GPC) of isoamylase-debranched starches. The results also indicated that different ae alleles had different effects on the amylose content of endosperm starches. Mature kernels of Oh43 ae-RWB-1 mutant showed tarnished and translucent phenotype characteristics of the ae genotype but contained endosperm starch with 21—22 % of amylose which was lower than that of the Oh43 normal counterpart. Both wx-B and wx-C (standard wx) genes have similar effect on structure and physicochemical properties of waxy starches of A632, B37, C105, Oh43, and W64A inbred lines.     

Comparison of Methods for Colorimetric Amylose Determination in Cereal Grains

Iodine‐based colorimetry remains the most widespread method for the determination of amylose content of starch. There are, however, many variants of this method, differing in solvent, temperature and method for converting absorbance to amylose content. Therefore, in thisstudy some of these methods were compared in order to identify which, if any, was most reliable. Australian commercial samples of wheat, rice, rye and oat were used. Flour was ground and starch was extracted. Half of the samples of flour and starch were defatted with n‐propanol. Defatted and full‐fat flours and starches were dissolved by six procedures: DMSO, urea‐DMSO, CaCl₂‐DMSO, hot NaOH, cold NaOH and HCl. The solutions were diluted, coloured with iodine and the absorbances determined at 535 and 620 nm. Amylose content was determined by reference to standards using 620 nm as the single wavelength or by two calculations based on the ratio between the two absorbances. Reliability was tested by repeating all tests on three non‐consecutive days with three replicates on each day. Defatted starch provided more consistent results than flour or non‐defatted starch. The most reliable dissolution procedure, with both the lowest standard error and least day‐to‐day variation, was cold NaOH. One of the ratio‐based calculations provided adequate correction for the non‐starch content of flour of wheat and rice, but not of the other two cereals, and the other ratio‐based calculation seldom provided realistic results. It is concluded that a cold NaOH dissolution procedure is suitable for reliable amylose determination on defatted starch of a range of cereals from a single wavelength and that it is also suitable for use on defatted flour of some species.     

Waxy Gene Factor and Residual Protein of Rice Starch Granules

Residual protein of rice starch granules prepared from milled rice by protein extraction with sodium dodecylbenzene sulfonate or alkaline protease treatment was related to amylose content. SDS-polyacrylamide disc gel electrophoresis confirmed that the major protein in nonwaxy starch granules was the Wx gene product with subunit molecular weight of about 60,000 daltons. The Wx gene product was absent in waxy starch granules.     

Cell Wall Degrading Enzymes and Proteases Improve Starch Yields of Sorghum and Maize

The effects of a commercial cell‐wall‐degrading enzyme (CWDE) complex and/or commercial protease on yield and quality of starch extracted from white regular sorghum (RSOR), white waxy sorghum (WSOR) and yellow maize (YMZ) were determined. A bifactorial experiment with a level of confidence of P<0.05 was performed to study the effect of CWDE and/or protease addition to coarsely ground grains for 4 h after 20 h of regular SO₂ steeping of whole kernels. Calcium oxide (CaO) or lactic acid were used to adjust the pH solution to 5.5 prior to grinding. YMZ yielded the highest amount of starch, whereas RSOR gave higher starch yield than WSOR. All enzyme treatments significantly (P<0.05) improved starch yields. Protease alone and CWDE plus protease treatments were the most effective treatments in terms of starch yields. These enzyme treatments allowed the recovery of more than 90% of the starch. The two types of sorghum starches contained higher amounts of residual protein and ash when compared with maize starch. The various enzymes treatments did not affect starch composition or Visco Analyser properties of each type of kernel. As expected, the WSOR starch had lower pasting temperature and retrogradation when compared with the regular starches.     

Properties of protease-treated maize starches

Commercially produced maize starches were treated with protease (Promod 25P) and their composition and properties were compared with untreated controls. It was found that, although protease treatment reduced the starch protein contents by 41%, 21% and 37% for the waxy, normal and amylomaize starches, respectively, it also caused some pits on the granule surfaces, which were evident by scanning electron microscopy (SEM), but no obvious decrease in granule dimensions (Coulter Counter Multisizer). The protein extraction was associated with decreases in starch lipid content by 42%, 40% and 45% (waxy, normal and amylomaize starches, respectively) and a decrease in total amylose content (30.7–26.0%) for the normal maize starch. The gelatinisation parameters of the starches by differential scanning calorimetry (DSC) in water, 0.001 M HCl or NaOH were less obviously affected by protease treatment in common with the swelling factors at 80 °C. The amount of α-glucan leached by the swollen (80 °C) granules was, however, increased by the protease treatment by factors of 3.8, 1.4, and 1.1, for the waxy, normal and amylomaize starches, respectively. Although proteases provide a useful tool for the purification of native starches, commercial protease preparations need to be controlled in terms of amylase content to prevent modifications to starch structure and properties during industrial processing.     

A Novel Type of Corn Starch from a Strain of Maize. Part 3. The Properties of the Starch from Amylo-waxy Maize and Its Hybrids with Waxy Maize

A pair of hybrids were produced by alternative crossbreeding of waxy maize with amylo-waxy maize which had double recessive genes, amylose extender (ae) and waxy (wx). Investigation of the starches proved that they did not coincide with waxy starches. Hidden amylose extender genes also participated in the starch produced in the endosperm of hybrid maize. Amylo-waxy starch looks promising as a new material of food industry.     

Comparison Between Granular Starch Hydrolyzing Enzyme and Conventional Enzymes for Ethanol Production from Maize Starch with Different Amylose: Amylopectin Ratios

The effect of enzyme treatments (granular starch hydrolyzing and conventional enzymes) and different amylose: amylopectin ratios of maize starch on ethanol production was evaluated. For starch treatments, amylose: amylopectin ratios were prepared by mixing commercially available Hylon VII (70% amylose and 30% amylopectin) and Amioca (˜100% amylopectin) starches. For maize treatments, waxy, high amylose and regular dent hybrids were used to represent varying amylose: amylopectin ratios. All hydrolyses followed by fermentations were conducted at 15% solids content. Differences were observed in ethanol yields among granular starch hydrolyzing and conventional enzymes. Differences also were observed in ethanol yields between different amylose: amylopectin ratios for pure starch and maize samples. For starch samples, final ethanol concentrations varied from 2.2 to 9.1% (v/v) for fermentation with granular starch hydrolyzing enzyme and from 6.7 to 9.3% (v/v) for conventional enzymes. Higher ethanol concentrations were observed for Amioca starch for both enzymes. For maize samples, final ethanol concentrations were highest for waxy maize for both granular starch hydrolyzing (8.2%, v/v) and conventional (8.2%, v/v) enzymes. Lowest ethanol concentrations were observed for high‐amylose maize samples for granular starch hydrolyzing (6.3%, v/v) and conventional (5.2%, v/v) enzymes.     

酶辅助碱法提取龙眼核淀粉的结构与理化性质研究

目的:明确酶辅助碱法提取的龙眼核淀粉的结构及理化性质。方法:以龙眼核为原料,碱性蛋白酶为酶解剂提取龙眼核淀粉,研究了其化学组成、粒径、微观形貌、晶体结构、透明度、溶解度及膨润力、热力学特性及糊化特性等。结果:100 g龙眼核淀粉中含有82.64 g淀粉、0.68 g粗脂肪、0.46 g粗蛋白和1.06 g灰分,其直链淀粉含量为36.37%;其颗粒多呈椭圆形或不规则多边形,平均粒径为11.21 μm,结晶结构为A型,近程有序程度较高;1%（w/v）龙眼核淀粉糊的透光率为4%;溶解度和膨润力均随温度上升而增大,90 ℃时分别为11.26%和15.93%;龙眼核淀粉具有较高的糊化起始温度（70.4 ℃）、峰值温度（75.9 ℃）、结束温度（82.1 ℃）和糊化焓（13.7 J/g）,其热糊稳定性较差,冷糊稳定性较好。结论:酶辅助碱法提取的龙眼核淀粉直链淀粉含量高,蛋白质含量下降明显,颗粒小,不易糊化。     

Chain‐length Distribution Profiles of Amylopectin Isolated from Endosperm Starch of Waxy and Low‐amylose Bread Wheat (Triticum aestivum L.) Lines with Common Genetic Background

Large A‐type endosperm starch granules were isolated from near‐isogenic waxy and non‐waxy lines and low‐amylose mutant lines of bread wheat with a common genetic background. The amylose contents of A‐type starch ranged from 2.6% to 23.6%. Amylopectin was isolated by concanavalin A (Con A) precipitation from the isolated starch. The λ_max (range: 532‐538 nm) and blue values at 680 nm (range: 0.026‐0.037) of the iodine‐amylopectin complex were not significantly different among the isolated amylopectins, indicating that amylopectins from non‐waxy and low‐amylose lines did not contain such long chains as amylose or extra‐long chains of amylopectin affecting iodine complex properties. Chain‐length distribution profiles measured by both high‐performance size‐exclusion chromatography (HPSEC) and high‐performance anion‐exchange chromatography (HPAEC) showed that the amylopectin structures of these lines were indistinguishable from each other. Extra‐long chains were not detected in the amylopectins by HPSEC measurement. The side‐chains measured by HPAEC were classified into four groups according to their degree of polymerization (DP), and the proportion of each group were in the following ranges: DP 6‐12, 26.5‐27.5%; DP 13‐24, 43.6‐44.1%; DP 25‐36, 13.6‐14.2%, and DP 37‐60, 11.0‐11.7%. The alleles on the Wx‐D1 locus, i.e., Wx‐D1a, Wx‐D1d, Wx‐D1f, and Wx‐D1g, responsible for granule‐bound starch synthase (GBSS I) biosynthesis had no influence on the properties of iodine‐amylopectin complex and the chain‐length distribution profiles of amylopectin.     

Characterization of Pinhão Starch. Part I. Extraction and Properties of the Starch Granules

The starch fraction of Pinhão, the seeds of Pinheiro do Paraná (Araucaria angustifolia, Bert, O. Ktze) was easily extracted and represents 21.87% of total weight. The low content of protein, fat and ash accounts for the efficiency of the extraction and purification. Pinhão starch shows an amylose level and strong anionic characteristic similar to corn starch. The round-shaped granules are smaller than those of cassava and similarly resistent to DMSO solubilization, what suggests a very homogenous granular structure.     

Characterization of Endosperm Starch from High-amylose Mutants of Rice (Oryza sativa L.)

The structure and properties of endosperm starch from high-amylose mutants of rice were examined. The starch in the mutants was characterized by a higher content of amylose and loosely branched amylopectin with longer chains compared with non-mutant starch. The starch granules in the mutants showed high temperatures of gelatinization and a type B pattern in X-ray diffractometry. These properties were similar to those of amylose-extender (ae) starch of maize. The effect of locations where rice plants were grown on the endosperm starches of one high-amylose mutant and a non-mutant was also investigated.     

Comparison of Protein Extraction Solutions for Rice Starch Isolation and Effects of Residual Protein Content on Starch Pasting Properties

Protein extraction solutions such as aqueous solutions of sodium hydroxide (0.1 and 0.2 %), sodium lauryl sulfate (SLS, 1.2 %) containing sodium sulfite (0.12 %), and dodecylbenzene sulfonate (DoBS, 1.2 %) containing sodium sulfite (0.12 %) were compared in their protein removal efficiencies during isolation of starch from a rice flour (Ilpumbyo, a nonwaxy Korean rice variety). In addition, the pasting properties of the isolated starch was compared. More than 80 % of the flour protein was extracted in 1 h by stirring the dispersion (1:3, w/v) at room temperature. Repeating the extractions (1 or 2 h for each step) with fresh solution significantly increased the protein removal efficiency. When the extraction in 0.2 % NaOH was repeated four times (1 h for each step) at 25°C, the residual protein content in the isolated rice starch was 0.9 % (DB), equivalent to 86 % removal of the rice protein. Raising the extraction temperature slightly increased the protein solubility, but starch loss also became significant. Among the solutions, DoBS was most effective in removing rice protein whereas SLS was least. The residual protein content had a critical role in determining the pasting characteristics of the isolated starch, showing a negative correlation to the peak viscosity of the starch paste, but a positive correlation to the pasting temperature.     

Effect of Gene Dosage at High Amylose Loci on the Properties of the Amylopectin Fractions of the Starches

The effect of gene dosage at the amylose-extender (ae) locus of maize and the rigosus-a (r_a) locus of peas on the amylopectin fraction of endosperm and cotyledon starch were examined. Amylopectin fractions were isolated by gel filtration, butanol complexing and a combination of the two methods. In general, increasing doses of the recessive allele at either locus resulted in amylopectin fractions with iodine spectra with absorption maxima at higher wavelengths and greater absorptivity. Starches from endosperm of differing ae dosage but homozygous waxy were found to contain no amylose by gel filtration. By debranching with pullulanase, these starches were shown to have increasing average chain length with increasing ae dosage. Additional data suggested that increasing dosage at the ae locus (regardless of the genotype at the waxy locus) or the r_a locus resulted in amylopectin with increasing linearity. In addition, short chained amylose (approximately 100 glucose units) was observed in all ae genotypes in a homozygous Waxy background and all r_a genotypes.     

烤烟烟叶淀粉的提取工艺优化

为揭示烤烟淀粉的结构与特性,降低烤烟的淀粉含量,采用响应面分析法(RSM)对鲜烟叶淀粉的提取条件进行了优化。结果表明:烤烟烟叶淀粉提取最优条件为:亚硫酸浓度0.19%,液固比为7,浸提时间16 h;淀粉中脂肪含量为0.61%,蛋白质含量为3.73%,灰分为1.59%,直链与支链结构比例约为3∶7。