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1.
To investigate the chemical heterogeneity of humic acids (HAs), we applied two-dimensional (2-D) electrophoresis to HAs from a compost and two types of soils. In this method, HAs are first separated by isoelectric focusing (IEF) and then separated by polyacrylamide gel electrophoresis (PAGE). IEF and PAGE were carried out in the presence of 7?M urea. Upon 2-D electrophoresis of HAs, dark-colored substances were spread out across the gel mainly in the isoelectric point (pI) range of 3.0–4.5. Green fluorescence was observed in the smaller molecular size region of the gel, especially in the pI range of 3.0–4.5, and the most intense fluorescence was found at the moving front. The gels were divided into 36 sections, and then HA constituents were extracted from the individual sections and recovered by precipitation with acid. The distribution of organic carbon (C) among the gel sections coincided with that of the dark-colored substances on the gel. The total C recoveries were only 43–50%, suggesting that a considerable amount of HA constituents was lost during the extraction from the gels and purification. High-performance size-exclusion chromatography confirmed that the constituents of HAs were separated based on their molecular sizes by PAGE. The measurement of diffuse reflectance infrared Fourier transform (DRIFT) spectra indicated that the chemical properties of the HA constituents differed depending on the position on the gels and were affected by the molecular size rather than the pI. The fractions of the compost HA were characterized by higher proportions of aliphatic, proteinous and polysaccharide moieties and by the presence of lignin-derived structures. For the soil HAs, the fractions were characterized by a high proportion of the carboxyl group and a low proportion of aliphatic moieties. The proportion of proteinous and polysaccharide moieties in the fractions of soil HAs decreased with decreasing molecular size. The chemical properties of the green fluorescent substances remained unclear, since there was not enough of the substances to measure the DRIFT spectra. The present study showed that 2-D electrophoresis in the presence of concentrated urea offers an effective method for fractionating and isolating the constituents of HAs.  相似文献   

2.
Humic acids (HAs) from four soils were fractionated by size exclusion chromatography (SEC) on Sephadex G-75. Three fractions were obtained in all humic acids, collected and assayed by Polyacrylamide gel electrophoresis (PAGE). The unfractionated HA from each soil was used as reference. Each chromatographic fraction formed one electrophoretic zone corresponding closely to one band of the reference sample with some admixture of the fractions preceding or following. The results indicate that fractionation of HAs by tandem SEC-PAGE can be successfully used for obtaining fractions of reduced polydispersity and different electrophoretic mobilities. Pyrolysis/methylation-gas chromatography-mass spectrometry of the full size preparations of HA and fractions with exactly defined molecular size and electrophoretic mobility showed a different distribution in humic components, mainly lipids, lignin derivatives and N-containing compounds.  相似文献   

3.
Polyacrylamide gel electrophoresis in combination with size-exclusion chromatography (SEC–PAGE) has been used to obtain stable electrophoretic fractions of different molecular size (MS) from chernozem humic acids (HAs). Three-dimensional fluorescence charts of chernozem HAs and their fractions have been obtained for the first time, and all fluorescence excitation–emission maxima have been identified in the excitation wavelength range of 250–500 nm. It has been found that fractionation by the SEC–PAGE method results in a nonuniform distribution of protein- and humin-like fluorescence of the original HA preparation among the electrophoretic fractions. The electrophoretic fractions of the highest and medium MSs have only the main protein-like fluorescence maximum and traces of humin-like fluorescence. In the electrophoretic fraction of the lowest MS, the intensity of protein-like fluorescence is low, but the major part of humin-like fluorescence is localized there. Relationships between the intensity of protein-like fluorescence and the weight distribution of amino acids have been revealed, as well as between the degree of aromaticity and the intensity of humin-like fluorescence in electrophoretic fractions of different MSs. The obtained relationships can be useful in the interpretation of the spatial structural organization and ecological functions of soil HAs.  相似文献   

4.

Purpose

The fractionation of soil humic acids (HAs) according to their hydrophobicity is a common procedure in the study of this polydispersed complex natural mixture, so that reversed-phase high-performance liquid chromatography (RP-HPLC) is used resulting in humic components of differing hydrophobic/hydrophilic properties. However, a comparative study of the hydrophobicity of fractions isolated from different soil HAs have not been addressed so far.

Materials and methods

The RP-HPLC with online absorbance detection was used for analysis of International Humic Substances Society soil standard HAs, chernozem soil HAs, and their electrophoretic fractions A, B, and C?+?D, obtained by tandem size exclusion chromatography–polyacrylamide gel electrophoresis. The strong relationship between hydrophobicity, electrophoretic mobility (EM), molecular size (MS), specific absorbance at 280 nm and aromaticity of HAs fractions was found.

Results and discussion

Independently of soil HAs genesis fraction A with lowest EM and highest MS is essentially more hydrophobic (60–73 % of the fraction amount remained adsorbed on the RP column) than medium EM and MS fraction B (33–47 % of the fraction amount remained adsorbed on the RP column). The lowest hydrophobicity belongs to fraction C?+?D with highest EM and lowest MS.

Conclusions

The most hydrophilic aromatic fraction C?+?D seems to have been bound with other mostly aliphatic hydrophobic fractions A and B through non covalent (possibly hydrogen) bonds. These data could be relevant to better understanding the overall makeup of soil HAs and their structural organization.  相似文献   

5.
Tandem size-exclusion chromatography and polyacrylamide gel electrophoresis were used for obtaining stable fractions of different molecular sizes and electrophoretic mobilities from chernozem humic acids (HAs). The obtained fractions were analyzed using solid-phase 13C NMR. The tendencies of the changes in the aromatic and aliphatic components of the HA fractions with different molecular sizes and electrophoretic mobilities were experimentally revealed. The aromatic-to-aliphatic carbon ratio Carom (165–108 ppm)/Caliph (108-0 ppm) was used for comparing the degrees of aliphaticity and aromaticity of the HA macromolecules. This ratio increased by more than five times when going from the high-molecular-weight to the low-molecular-weight fractions and largely determined their hydrophilic properties. The obtained results can be useful for the interpretation of the structural organization and the ecological functions of soil HAs and their fractions.  相似文献   

6.
The use of organic amendments requires an adequate control of the chemical quality of their humic acid (HA)-like fractions and of the effects that these materials may have on the status, quality, chemistry and functions of native soil HAs. In this work, the compositional, functional and structural properties of the HA-like fractions isolated from a liquid swine manure (LSM), a municipal sewage sludge (SS), and two municipal solid waste composts (MSWCs) were evaluated in comparison to those of HAs isolated from three unamended soils and from the corresponding soils amended with LSW, SS, and MSWC at various rates in three field plot experiments conducted in Minnesota, USA. With respect to the unamended soil HAs, the HA-like fractions of the three amendments featured a greater aliphatic character, a marked presence of proteinaceous, S-containing and polysaccharides-like structures, an extended molecular heterogeneity, small organic free radical contents and a small degree of humification. The MSWC-HAs featured a larger degree of humification than LSM-HA and SS-HA. The three amendments affected in different ways and by various extents the compositional, structural and functional properties of soil HAs depending upon the nature, origin and application rate of the amendment. In general, the data obtained suggested that proteinaceous, S-containing and aliphatic structures contained in HA-like fractions of organic amendments were partially incorporated into native soil HAs.  相似文献   

7.

Purpose

Anion exchange resin has been shown to be efficient for adsorption of humic acid (HA). However, the mechanism of the effect(s) of the molecular weight (MW) of HA on the adsorption by anion exchange resin needs to be further explored. The objective of this work was to investigated the effect(s) of HA MW on their adsorption and desorption behavior by interacting with anion exchange resin.

Materials and methods

HA fractions with different MW were obtained by ultrafiltration and characterized. For adsorption kinetic study, the magnetic polyacrylic anion exchange resin (NDMP) was shaken with each HA fraction. The amounts of HA in the solutions at different sampling times were obtained by measurement of the total organic carbon. For isotherm study, batch experiments were performed to obtain the isotherms of each HA fractions. The effects of pH value and the regeneration efficiency for HA fractions with different MW were all investigated.

Results and discussion

The characterization reveals that aromaticity of HA increases as the MW rises, while the negative charge follows the order: HA3 (2,500–1,000 Da)?>?HA4 (>10,000 Da)?>?HA2 (1,000–2,500 Da)?>?HA1 (<1,000 Da). The adsorption amount of HA increases as the negative surface charge rises. Thus, HA3 has the largest adsorption amount by NDMP resin among HA fractions, while the adsorption amount of HA1 is the smallest. HA2 adsorption could be modeled by Freundlich equation, suggesting the interaction between HA2 and NDMP is a heterogeneous process. However, the adsorption isotherms of HA3 and HA4 were both better described by Langmuir equation than Freundlich equation. In addition, the resin fouling follows the order: HA2?>?HA1?>?HA4?>?HA3.

Conclusions

The aromaticity of HA freactions increases as MW rises. HA fractions with 1,000?–?2,500 Da have the largest surface negative charge, while the fractions with MW lower than 1,000 Da are almost electroneutral. The adsorptions of HA fractions onto NDMP suggest there is little effect of size exclusion on adsorption, and the adsorption behavior mainly depends on the negative charge of HAs.  相似文献   

8.
The distribution of selected elements in individual fractions of organic matter from anthropogenically contaminated soils was investigated. The attention was paid especially at Hg. Furthermore, contents of S, Mg, Mn, Fe, Cu, Zn and Pb were also measured. The decomposition of organic matter to particular fractions was carried out by the resin DAX-8. Ten soil samples were collected, and the Advanced Mercury Analyzer (AMA-254) was used for the determination of the total Hg content. The two highest Hg values reached up to the concentration 10.5 mg kg?1, and in the highest one, it was almost 29 mg kg?1. In each extract, mercury was measured by inductively coupled plasma mass spectrometry (ICP-MS), for other elements, inductively coupled plasma optical emission spectrometry (ICP-OES) was applied. Results of the analysis show that the Hg content bound to the humic acids is inversely proportional to the content of Mg, Mn, Fe and Cu. However, this dependence was not confirmed by the samples with the mercury content above 10 mg kg?1. In the case of fulvic acids, the relationship between Hg and S was observed and has again an inverse character.  相似文献   

9.
We investigated the optical absorption properties (A 400/C and A 600/C values), elemental composition, weight-averaged molecular weight (Mw), and liquid-state carbon-13 (13C) nuclear magnetic resonance (NMR) spectroscopy of DAX-8 adsorbed fulvic acid fractions (DAX-8 FAs) in various soils (n?=?36) to establish the general trends in their chemical characteristics. We also compared our results with those of humic acids (HAs) obtained from the same soils used in this study and other published data to discuss the differences between them. Our results clearly indicate that DAX-8 FAs with high carboxyl C content have small Mw and low hydrogen to carbon (H/C) ratio. In addition, DAX-8 FAs with high carbonyl C and alkyl C content have high A 400/C values and H/C ratios, respectively. These results strongly suggest that DAX-8 FAs with high aliphatic chemical properties have low carboxyl C content, large Mw and high H/C ratio. There are significant differences in chemical characteristics among the various soil types in almost all data; e.g., DAX-8 FAs from Podzols showed significantly higher A 400/C values, larger Mw, higher carbonyl C and O-aryl C content than those obtained from Andosols. However, these two C functional groups are relatively low values in the total C content among all soil types, suggesting that DAX-8 FAs may exhibit only small differences in chemical properties among the various soil types. The comparison with published data regarding soil HAs shows that DAX-8 FAs are characterized by smaller Mw; lower C, H, and nitrogen (N); higher oxygen (O) content; higher O/C; lower O/H ratios; higher carboxyl C content; and lower aryl C content in total C.  相似文献   

10.
Purification of gliadin subclasses has been difficult since they share many biochemical and physicochemical properties. In this report, the optimization of a preparative electrophoretic method to fractionate gliadins is described. Separation was performed in preparative 7% polyacrylamide gels at pH 3.1. The separation performance was tested using analytical electrophoresis at pH 3.1 and capillary electrophoresis. Preparative gels of different lengths were employed. Using 5-cm preparative gels, several fractions of alpha-, beta-, and gamma-gliadins and fast-mobility and slow-mobility omega-gliadins were collected in 40 h of separation. Resolution was maintained at a protein load of up to 30 mg in each run. The highest efficiency of recovery was achieved using aluminum lactate as the collecting buffer. Fractionation with 10 cm in length gels improved resolution but increased operation times. Gels of 2 cm in length did not separate alpha/beta- and gamma-gliadins efficiently but were useful to separate the two main fractions of omega-gliadins in shorter times. In conclusion, preparative electrophoresis at low pH allowed the separation of alpha-, beta-, gamma-, and omega-gliadin fractions from crude material under nondenaturing conditions.  相似文献   

11.
Humic acids (HAs) represent heterogeneous and polydisperse mixture of molecules that differ in their chemical structure, composition, and functional properties. Fractionation of HAs is of key importance for understanding their interactions with various organic and inorganic compounds, for studying their physiological activity, and for predicting their behavior in natural environments and agroecosystems. Existing fractionation methods are rather laborious and time consuming, which limits their application in fundamental science and industry. It is shown that fractionation of humic acids with ammonium sulfate ensures their preparative separation with respect to (a) hydrophobicity, (b) molecular size, and (c) charge dependent on the amount of functional groups. Salting out at the lowest and highest degrees of saturation with ammonium sulfate, upon which precipitation of the molecules occurs, makes it possible to separate humic acids into functionally different high-molecular-weight/hydrophobic and low-molecular-weight/hydrophilic fractions. The first fraction is characterized by a lower electrophoretic mobility than the second fraction. The weight percentage of the components coagulated at the lowest degree of salt saturation can be used as a quantitative parameter for comparing hydrophobic properties of humic acids. Salting out is recommended as a fast, simple, and cheap alternative to chromatographic methods for preparative separation of humic acids if large amounts of functionally different fractions need to be obtained.  相似文献   

12.

Purpose

The present study focuses on a compositional characterization of the humic acid (HA) fraction of several wetland soils using solid-state 13C NMR spectroscopy. The data were analysed using the molecular mixing model (MMM), based on an empirical approach by Nelson and Baldock. The compositional data from HAs obtained with this model were used to obtain a wider assessment of the process of humification from comparison of total soil wetland organic matter composition and HA composition.

Materials and methods

Twenty samples of humic acids (HAs) isolated from a Mediterranean semiarid wetland (‘Tablas de Daimiel’, central Spain) were studied using elemental analysis and cross polarization magic angle spinning (CPMAS)13C nuclear magnetic resonance (NMR) spectroscopy. The NMR data were analysed with the molecular mixing model (MMM) considering up to six generic components (carbohydrate, protein, lignin, lipid, char and ‘carbonyl’). HAs are considered a conceptual mixture of these model components, and the MMM determines the proportions of the characteristic biomolecules contributing to HA composition.

Results and discussion

The composition of the HAs under study depends on local factors such as site vegetation and occurrence of fire. Correlations between the proportions of the six generic components and further comparison with those determined for the unfractionated OM (whole sample, WS), gave information on HA origin and humification mechanisms. In particular, the proportions of char and carbohydrate (R 2 0.637) and contents of lignin and protein (R 2 0.471) in the HAs were negatively correlated (P < 0.05). Significant correlations (R 2 0.439) also existed for char contents in whole sample (WS) compared to HA, and for carbohydrates in WS compared to HA (R 2 0.558). Char proportion grew in HA with respect to the WS, and carbohydrates dropped to a half on average in HA compared to WS.

Conclusions

Two different humification mechanisms could be identified for no-fire and fire areas. In the former, HA-char was preserved selectively from char in the sample, whereas in the latter, char was newly formed by fire effect.
  相似文献   

13.
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.  相似文献   

14.
Water extracts were obtained from four types of soils (Brown Lowland soil, Yellow soil with manure application for 6 years, non-allophanic Andosol, and allophanic Andosol), and the organic matter in the water extracts was fractionated according to the solubility in acid and adsorption onto polyvinylpyrrolidone (PVP). For the water extracts and their fractions, the amounts of organic C, total N, and anthrone-reactive C (ARC) were analyzed, and high performance size exclusion chromatography (HPSEC) was carried out. The PVP-non-adsorbed fulvic acid (FA) fraction accounted for the largest proportion of the total water-soluble organic C, ranging from 52% for the Yellow soil to 96% for the allophanic Amdosol, followed by the PVP-adsorbed FA and humic acid (HA) fractions. The water extract of the allophanic Andosol was characterized by the lack of HA fraction and a small proportion of PVP-adsorbed FA fraction. For all the water extract samples, more than 90% of the ARC was recovered in the PVP-non-adsorbed FA fraction. The proportion of ARC in the total organic C in the fraction was also highest in the PVP-non-adsorbed FA fraction. The molecular weight (MW) of the humic substances (HS) at peak maximum was estimated at 1,300 Da for the water extracts and their fractions from the Brown Lowland soil and non-allophanic Andosol samples by HPSEC using polyethylene glycols as MW standards. Manure application increased the MW of HS in the HA and PVP-adsorbed FA fractions. On the other hand, only a small amount of HS was found in the water extract of the allophanic Andosol by HPSEC.  相似文献   

15.
Although condensed aromatic components are considered to be one of the major structural units of soil humic acids (HAs) and to be responsible for the dark colour of HAs, their amount and composition remain largely unknown. In ruthenium tetroxide oxidation (RTO), condensed aromatic components are detectable as their degradation products, mainly benzenepolycarboxylic acids (BPCAs). We applied this technique to soil HAs with various degrees of humification (darkening). The yields of water‐ and dichloromethane‐soluble products from HAs upon RTO after methylation ranged from 210 to 430 mg g−1 and 10–40 mg g−1, respectively. Eight kinds of BPCAs with two to six carboxyl groups, and seven kinds of BPCAs with additional side chains (tentative assignment) were obtained as methylated counterparts. The yield of each BPCA and the sum of the yields of BPCAs (12–85 mg g−1 HAs) increased with increasing degree of humification and aromatic C content. The compositions of BPCAs indicated that the degree of condensation was greater in the HAs with greater degrees of humification. The sum of the yields of aliphatic compounds ranged from 0.1 to 6.5 mg g−1, and decreased with increasing degree of humification. The C12 to C30 monocarboxylic acid methyl esters accounted for > 56% of the aliphatic compounds assigned, which may be present mainly as end alkyl groups in the HA molecules. We also obtained the methylated counterparts of C14 to C24 dicarboxylic acids; these were possibly derived from polymethylene bridges between adjacent aromatic rings.  相似文献   

16.
Andosols are characterized by an abundance of black humic acids (HAs) belonging to Type A with a high content of aromatic carbon (C) in particular condensed aromatic C. Black HAs are also observed in other soils, such as Chernozems and the subsoil of paddy field, and extracted after washing with an acid or using chelating agent such as sodium pyrophosphate (Na4P2O7). However, contribution of condensed aromatic structures to those soil HAs are unknown. To obtain the information about C skeletal structures of black HAs in soils other than Andosols, HAs were obtained from 2 Chinese Chernozem samples, 2 subsoil samples from Japanese paddy fields (Fulvisols), and a Rendzina-like soil (Cambisols) as well as an Andosol sample (reference) by successive extraction with 0.1 M NaOH (HAs1) and 0.1 M Na4P2O7 (HAs2), and 13C nuclear magnetic resonance and X-ray diffraction 11-band profile analyses were applied. In the black HAs2 from the non-Andosol samples, the proportion of C present as aromatic C, size of C layer planes, and relative C layer plane content ranged from 52 to 59%, 0.48 to 1.92 nm (mean size, 0.76–0.91 nm), and 58 to 100 AU (arbitrary unit) mg?1, respectively, with a positive correlation between total C layer plane content and the degree of humification. Those ranges were similar to the distribution ranges of Andosols HAs1 reported by our previous study.  相似文献   

17.
Quantitative and qualitative analysis of uncooked zein and kafirin fractions were performed through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretic profiles. Kafirins and zeins present the same oligomer and monomer compositions with the exception of a 66 kDa oligomer that is only present in kafirins. The quantitative analysis showed differences between zein and kafirin. The composition of each oligomer was established via preparative SDS-PAGE. Part of the cooked oligomers resists reduction; the presence of those oligomers could be related to the decrease on protein digestibility with the cooking process.  相似文献   

18.
Purpose

Biochar is one of the most widely used ameliorants for soil amendment, which is known as factor which rises crop yields and levels of soil biological activity. Nowadays, it is under investigated how biochar application affects the dynamics of the humic components and whole soil organic matter (SOM) and the processes of its alteration. This investigation is aimed to evaluate the influence of biochar on the content, composition, and transformation of humic acids (HAs) as the main component of the SOM.

Materials and methods

The incubation experiment was carried out on three Podzol Antric soils, with varying amounts of initial total organic carbon. The incubation time was 90 days, using biochar gravimetric doses of 0.1 and 1.0%. The biochar was produced by fast pyrolysis of birch and aspen wood at 550 °С. Humus composition was analyzed for the organic matter fractions extracted with 0.1 M NaOH (containing HAs 1 + fulvic acids (FAs) 1) and 0.1 M Na4P2O7 (containing HAs 1 + FAs 1 + HAs 2 + FAs 2). Isolated HAs were characterized for their elemental composition (C, N, H, and S) and molecular composition with the use of solid-state 13C nuclear magnetic resonance (13C-NMR) techniques.

Results and discussion

We found that 0.1% of biochar amendment does not influence SOM mineralization, but 1.0% of biochar increases the mineralization by 15–18%. This process is accompanied by changes in the composition and properties of the HS. The increased proportion of HA aromatic fragments in biochar indicates an increasing of their stability. However, in soils with high humus content and a significant amount of insoluble matter, the processes of mineralization and the growth of HAs are taking place simultaneously. The replenishment of HAs could be the outcome of both the intensification of the transformation processes (mineralization and humification) of the more sustainable insoluble matter compounds and the humification of the biochar itself.

Conclusions

The influence of biochar on humification in Podzol Antric soils was revealed on the basis of incubation experiment. Both negative and positive changes under biochar in HS system were demonstrated. The active decrease of humus total contents and also the labile HS ought to qualify as negative changes. The increase of HA chemical maturity that leads to the stability of humus in whole as well as the intensive new HA formation thought to qualify as positive changes.

  相似文献   

19.
The polyacrylamide gel electrophoresis of chernozem humic acids (HAs) followed by ultraviolet detection (λ = 312 nm) has revealed a new highly fluorescent fraction that has the highest electrophoretic mobility and the lowest nominal molecular weight (NMW). The preparative isolation of the fraction has been performed using the multiple microfiltration of the same HA sample in a 7 M carbamide solution on a membrane with a nominal pore size of 5 kDa. Thirty ultrafiltrates with NMW < 5 kDa and different fluorescence maximums in the region of 475–505 nm have been prepared, as well as a nonfluorescent concentrate with NMW > 5 kDa. Fluorescence maximums at and below 490 nm have been noted only in the first four ultrafiltrates. All the ultrafiltrates have been combined into the fraction with NMW < 5 kDa, which has been successively passed through membranes of 3 and 1 kDa. Solutions of subfractions F 3–5 kDa, F 1–3 kDa, and F < 1 kDa with fluorescence maximums at 505, 488, and 465 nm, respectively, have been prepared. The F < 1 kDa subfraction with the lowest NMW had the highest fluorescence intensity. The distribution of the fluorescence maximums in the ultrafiltrates has indicated the presence of at least two groups of fluorophores and has confirmed the supramolecular organization of the extracted soil HAs.  相似文献   

20.
Three samples of Nekota (hard red winter wheat) were milled, and six mill streams were collected from each sample. The 18 mill streams were analyzed separately as well as recombined to form three patent flours. The methods of multistacking (MS)‐SDS‐PAGE and SDS‐PAGE were used to separate the unreduced SDS‐soluble glutenins and the total reduced proteins, respectively. The separated proteins were quantified by densitometry. The quantity of unreduced SDS‐soluble proteins was significantly different among the mill streams at the 4% (largest molecular weight polymeric glutenins) and at the 10 and 12% (smaller molecular weight polymeric glutenins) origins of the MS‐SDS‐PAGE gels. The quantities of total HMW‐GS, LMW‐GS, 2*, 7+9, and 5+10 subunits and the ratio of HMW‐GS to LMW‐GS in polymeric protein samples isolated using preparative MS‐SDS‐PAGE and in total reduced protein extracts were significantly different among mill streams. The quantities of HMW‐GS, LMW‐GS, 2*, 7+9, and 5+10 subunits from total reduced proteins were positively and significantly correlated with loaf volume. The quantities of glutenin subunits (both HMW‐GS and LMW‐GS) from unreduced SDS‐soluble proteins were positively or negatively correlated with loaf volume at the various MS‐SDS‐PAGE gel origins but the levels of correlation were not significant. These results showed that the glutenin protein composition was different among the various mill streams and demonstrated that electrophoretic analysis of the proteins in these fractions is a useful tool for studying the variation in functional properties of flour mill streams.  相似文献   

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