Affinity Chromatography with Immobilised Endoxylanases Separates TAXI- and XIP-type Endoxylanase Inhibitors from Wheat (Triticum aestivum L.)

An affinity-based purification procedure allowed the resolution of two distinct groups of endoxylanase inhibitors with different molecular structures and endoxylanase specificities from wheat wholemeal. The first group comprises the so-called Triticum aestivum L. Endoxylanase inhibitor (TAXI)-type proteins which are of approx. M_r 40 000 and occur in two different molecular forms. These inhibitors were removed from a concentrated cation exchange chromatography fraction from wheat wholemeal on a Bacillus subtilis endoxylanase affinity column. The second group of structurally different endoxylanase inhibitors, the so-called xylanase inhibiting protein (XIP)-type, of approx.M_r 29 000–32 000, with pI values varying between 8·8 and 9·2, was purified from the unbound fraction from the B. subtilis endoxylanase affinity column by chromatography on an Aspergillus niger endoxylanase affinity column followed by gel permeation chromatography. The XIP-type inhibitors are not active against the B. subtilis endoxylanase and were consequently not retained on the B. subtilis endoxylanase column. Further analysis of the XIP-type proteins by high-resolution cation exchange chromatography, SDS-PAGE and iso-electrofocusing, revealed several forms. They had similar endoxylanase specificities and N-terminal amino acid sequences.     

In-source pyrolysis-field ionization mass spectrometry and Curie-point pyrolysis-gas chromatography/mass spectrometry of amino acids in humic substances and soils

With the aid of in-source pyrolysis-field ionization mass spectrometry (Py-FIMS) and Curie-point pyrolysis-gas chromatography/mass spectrometry (cPy-GC/MS) in the conventional electron impact mode, characteristic signals of 23 amino acid standards were described. Thermal and mass spectrometric fragmentation pathways of these amino acids differed with each method and complemented each other. Pyrolysis products assigned by Py-FIMS extended the range of signals for N-containing compounds in humic substances and soil organic matter and gave marker signals for free amino acids and their subunits in proteinaceous materials. These characteristic signals were correlated with the amino acid content in N-rich humic fractions consisting of seven fulvic acids and eight humic acids. The selected marker signals reflected 25–84% of the variances of the molar distribution of acidic, neutral, neutral aromatic, and basic amino acids in the humic fractions. In addition, a well described agricultural soil (0.08% amino acid N) was spiked with a standard amino acid mixture (0.08 mg amino acid N 100 mg^-1 dry soil) and produced enhancements of the relative abundances of the corresponding amino acid signals. Moreover, for 27 samples of whole agricultural soils of widely different origins, soil types, and vegetations, 15 selected amino acid indicators were correlated significantly with -amino N (r=0.76^***) and total N (r=0.65^***).     

不同产地红天麻多酚组成及体外抗氧化活性研究

【目的】比较4个不同产地红天麻的多酚含量、组成及其抗氧化活性,为天麻资源的综合开发利用提供参考。【方法】以4个产地（陕西、云南、贵州和四川）的红天麻为原料,提取红天麻中的游离酚和结合酚,采用Folin Ciocaileu比色法测定总多酚含量,超高效液相色谱法（UPLC）分析其多酚组成。通过测定红天麻多酚对1,1-二苯基-2-三硝基苯肼（DPPH）自由基、羟自由基和2,2′-联氮双-（3-乙基苯并噻唑林-6-磺酸）二胺盐（ABTS）自由基的清除能力评价其体外抗氧化活性。【结果】4个产地红天麻的游离酚含量为371.57~442.62 μg/g,结合酚含量为65.34~83.10 μg/g,游离酚含量占总多酚的84.3%以上,约为结合酚的5.4倍。4个产地红天麻总多酚含量为436.91~525.72 μg/g,总多酚平均含量为486.27 μg/g,其中陕西汉中红天麻总多酚含量最高。4个产地红天麻中主要多酚组分为荭草素、表儿茶素、咖啡酸、肉桂酸、对羟基苯甲酸。体外抗氧化结果显示,4个产地红天麻均有一定的抗氧化活性,相同质量浓度结合酚的抗氧化活性高于游离酚。清除DPPH自由基的IC₅₀（达到50%抑制效果时的质量浓度）为1.35~9.62 mg/mL,清除羟自由基的IC₅₀为2.65~28.58 mg/mL,清除ABTS自由基的IC₅₀为1.01~7.11 mg/mL。抗氧化活性综合（APC）指数结果显示,不同产地红天麻游离酚的APC指数均明显低于结合酚,其中贵州大方红天麻结合酚的APC指数最高,为96.04%,抗氧化活性最强;陕西汉中红天麻结合酚的APC指数为74.91%,抗氧化活性次之。【结论】4个产地红天麻多酚组成均较为丰富,且具有较好的体外抗氧化活性。     

Variation in avenanthramide content in spring oat over multiple environments

Avenanthramides (AVNs), a group of phytochemicals which are unique to oats, provide health benefits through antioxidant activity and other bioactivities. In this study, we explored genotype-by-environment interactions and heritability for AVN concentrations in oats. Avenanthramide concentrations were quantified for 100 breeding lines and cultivars at three locations over two years. While year and environment had an influence on AVN concentrations, with the influence of year being more apparent than that of environment within a year, genotype had the largest impact on AVN concentration. All three major AVNs were found to be heritable. Two methods of calculating heritability on a line mean basis were used. The statistical method yielded heritability estimates of 0.34, 0.39, and 0.41 for AVN 2c, AVN 2p, and AVN 2f, respectively. By comparison the traditional plant breeding method yielded heritability estimates of 0.82, 0.88, and 0.89 for AVN 2c, AVN 2p, and AVN 2f, respectively, indicating that expected gains will be dependent on the scale of the breeding program, the number of target environments, and climate variability. The estimated heritabilities and the 11-fold range in AVN concentrations in the 100 genotypes studied provided evidence that variability for AVN concentration should allow breeding progress for higher AVN concentration.     

Determination of Mugineic Acid, 2′-Deoxymugineic Acid, 3-Hydroxymugineic Acid,and Their Iron Complexes by Ion-Pair HPLC and Colorimetric Procedures

Extract

Under iron (Fe)-deficient conditions like in calcareous and/or high pH soils, mugineic acid family phytosiderophores (MAs: mugineic acid (MA), 2′-deoxymugineic acid (DMA), 3-hydroxymugineic acid (HMA) etc.) are secreted from graminaceous plants and solubilize the slightly soluble Fe in soil as MAs-Fe complexes (Takagi 1976, 1993). Due to their high availability to higher plants (Roemheld and Marschner 1986), the behavior of MAs and their Fe complexes in the soil environment is of interest in connection with the iron nutrition of these plants.     

High Performance Size Exclusion Chromatography of Water Extract from Sewage Sludge-Soil Mixture

In order to investigate the degradation or synthetic process of organic matter in soil, some instrumental analyses on the water extract from soil have been recently performed. SCHAUMBERG et al. (1) who observed the changes in the chemical nature of water-soluble components of sewage sludge-soil mixtures by using infrared spectroscopy, supplied valuable information on the fate of sludge organic components in soil. The authors reported (2, 3) that the stabilization process of sewage sludge in soil can be monitored by the conventional gel chromatography method with Sephadex G-15 in water extracts from sewage sludge-soil mixtures.

The previous works (2, 3) showed that macromolecular compounds which cannot be separated by Sephadex G-15 gel were present in the water extracts from sewage sludge-soil mixtures. High performance size exclusion chromatography (HPSEC) offers the possibility to estimate the molecular size distribution of such macromolecules, because HPSEC allows rapid size separation and high resolution. In the size separation of water-soluble compounds by HPSEC, however, the charge exclusion and the hydrophobic interaction between solutes and the stationary phase of the column caused accelerated or retarded transportation of water-soluble compounds through the column, leading to the erroneous estimation of the molecular size (4).

In this study, the authors applied HPSEC to water extracts from sewage sludgesoil mixtures during their process of decomposition in soil to observe the change of the molecular size distribution of the components.     

高压脉冲电场辅助溶剂提取蛋黄卵磷脂效果及品质保证

为提高对蛋黄卵磷脂(PC)的提取率,该文利用高压脉冲电场(PEF)辅助有机溶剂来进行提取试验。试验选定了影响PC提取率的3个主要因素:PEF场强、脉冲数和提取溶剂含水率,并采用Box-Behnken试验设计对提取工艺进行优化;并且,通过中红外光谱扫描分析了经PEF处理前后的PC分子结构变化。结果表明:PC提取的最佳工艺条件为场强39kV/cm,脉冲数31,提取溶剂含水率9%,其PC提取率比无PEF处理的传统有机溶剂提取法提高了10.2%;PEF对PC分子结构并没有显著影响,证实了PEF对食品营养成分破坏少,是一种温和的处理方式。该研究可为PEF技术应用于蛋黄卵磷脂的提取提供参考。     

Properties of neutral phosphate buffer extractable organic matter in soils revealed using size exclusion chromatography and fractionation with polyvinylpyrrolidone

Abstract

A neutral phosphate buffer (NPB) extraction method has been used to estimate the amount of available N in soil. However, the properties of soil NPB-extractable organic N have not been fully elucidated. The purpose of the present study was to characterize the properties of organic matter in the NPB extracts of soils. The NPB extracts were obtained from three soil samples, and the organic matter in the extracts was separated into three fractions according to its solubility in acid and adsorption onto polyvinylpyrrolidone (PVP). High-performance size exclusion chromatography (HPSEC) with ultraviolet (UV) and fluorescence detections was applied to the NPB extracts and their fractions. The HPSEC analysis of the NPB extract revealed the presence of a single broad peak, irrespective of the detection methods. The broad peak was identified as humic substances using the on-flow measurements of UV absorption spectra and fluorescence emission spectra. Among the fractions, the PVP-non-adsorbed fulvic acid (FA) fraction accounted for the largest proportion of organic C or N in the NPB extract, followed by the PVP-adsorbed FA and humic acid (HA) fractions. The peak of humic substances was observed for all fractions using HPSEC with the on-flow measurement of UV absorption and fluorescence emission spectra. The molecular weight of the humic substances varied with each fraction. When the Coomassie Blue-reactive substances (CBRS) were quantified using a Bradford protein assay, they were detected in the NPB extract and almost half were distributed in the PVP-non-adsorbed FA fraction. However, humic substances were considered to be the main constituents of CBRS in the soil NPB extract because of their reactivity with Coomassie Blue and the absence of proteinaceous materials. Furthermore, an incubation experiment revealed that the organic matter available to microorganisms was included in the HA and PVP-non-adsorbed FA fractions. Based on the HPSEC analysis of the NPB extracts and their fractions, it was observed that the humic substances in the NPB extract, particularly in the HA and PVP-non-adsorbed FA fractions, were available to microorganisms.     

高效液相色谱法测定无核白葡萄赤霉素残留

为建立高效液相色谱法测定无核白葡萄果实中赤霉素含量（残留）的方法,样品采用冷甲醇萃取,超声波提取,离心后上清液经0.45 um 滤膜过滤,滤液用 HPLC 紫外检测器测定。色谱条件为以 A（0.05%磷酸水溶液）/B（乙腈）（A / B：80%/20%）为流动相,ZORBAX Extend C18柱,柱温为30℃,紫外检测器波长210 nm。结果表明：该法线性良好,相关系数r＞0.999,精密度在0.86%～3.43%范围之内,平均回收率89.83%。本方法快速、可靠、灵敏,可为测定无核白葡萄果实中赤霉素含量提供参考。     

水发海参中甲醛的气相色谱法测定

本文探讨了一种水发海参中游离甲醛的气相色谱测定方法。甲醛以2，4－二硝基苯肼衍生后，经OV－17柱分析，电子捕获检测器检测。方法灵敏度高，重现性好，且衍生产物稳定，室温下放置，4天内测定结果平行。