beta-lactoglobulin hydrolysis. 1. Peptide composition and functional properties of hydrolysates obtained by the action of plasmin, trypsin, and Staphylococcus aureus V8 protease.

beta-Lactoglobulin (betaLg) was subjected to limited hydrolysis by trypsin, plasmin, and endoproteinase from Staphylococcus aureus V8 (S.aur.V8) to degrees of hydrolysis (DH) of 1, 2, and 4%. The several hydrolysates had different peptide compositions (determined by reversed-phase HPLC and gel-permeation chromatography [GPC]). GPC under nondenaturing, denaturing, and denaturing plus reducing conditions showed that the peptides formed were linked by hydrophobic interactions or by disulfide bonds or were not linked at all. At very low protein concentration, some differences in emulsion-forming properties were observed: only the plasmin hydrolysates could form emulsions with a uniform particle-size distribution. The emulsions formed with S.aur.V8 hydrolysates had poor emulsion-stabilizing properties. Some hydrolysates showed increased foam-forming properties in comparison with the intact protein. All foams formed were stable. Overall, the plasmin hydrolysate (DH4) contained relatively much larger molecules and/or hydrophobic molecules. Many molecules were disulfide-linked peptides. This hydrolysate also had the best functional properties.     

Effect of physicochemical conditions on peptide-peptide interactions in a tryptic hydrolysate of beta-lactoglobulin and identification of aggregating peptides

The objective of this study was to characterize the changes in peptide solubility resulting from changing some physicochemical conditions in a tryptic hydrolysate of beta-lactoglobulin (beta-LG). The turbidity (500 nm) of a 1% solution of tryptic peptides was measured at pH 3-10, at 5, 25, and 50 degrees C, in the presence of different salt concentrations (0, 0.5, and 1 M NaCl), in the presence of denaturing and reducing agents (6 M urea, 5% SDS, or 5% beta-mercaptoethanol), and under an electric field (isoelectric focusing). The results reveal an increase in turbidity of the peptide solution at pH 4, but a slight increase in turbidity was also observed at pH 8, which is attributable to peptides linked by disulfide bridges. The effect of temperature and ionic strength on the turbidity occurring at pH 4 indicates that mainly hydrophobic interactions are involved in the aggregation process. The material in the precipitate at pH 4 was identified as the peptides beta-LG 1-8, 15-20, and 41-60 and non-hydrolyzed alpha-lactalbumin. These results suggest that a limited number of peptides are involved in the aggregation process observed at pH 4, some of which having bioactive (beta-LG 15-20, ACE inhibitor, and opioid) or emulsifying properties (beta-LG 41-60). Aggregation of these peptides at acidic pH indicates that a simple acidification step could represent an easy process for isolating peptidic fractions enriched in bioactive or functional peptides.     

Hydrophobicity,Solubility, and Emulsifying Properties of Enzyme-Modified Rice Endosperm Protein

Rice endosperm protein was modified to enhance solubility and emulsifying properties by controlled enzymatic hydrolysis. The optimum degree of hydrolysis (DH) was determined for acid, neutral, and alkaline type proteases. Solubility and emulsifying properties of the hydrolysates were compared and correlated with DH and surface hydrophobicity. DH was positively associated with solubility of resulting protein hydrolysate regardless of the hydrolyzing enzyme, but enzyme specificity and DH interactively determined the emulsifying properties of the protein hydrolysate. The optimum DH was 6–10% for good emulsifying properties of rice protein, depending on enzyme specificity. High hydrophobic and sulfhydryl disulfide (SH-SS) interactions contributed to protein insolubility even at high DH. The exposure of buried hydrophobic regions of protein that accompanied high-temperature enzyme inactivation promoted aggregation and cross-linking of partially hydrolyzed proteins, thus decreasing the solubility and emulsifying properties of the resulting hydrolysate. Due to the highly insoluble nature of rice protein, surface hydrophobicity was not a reliable indicator for predicting protein solubility and emulsifying properties. Solubility and molecular flexibility are the essential factors in achieving good emulsifying properties of rice endosperm protein isolates.     

Hydrolysis of whey protein isolate with Bacillus licheniformis protease: aggregating capacities of peptide fractions

In a previous study, peptides aggregating at pH 7.0 derived from a whey protein hydrolysate made with Bacillus licheniformis protease were fractionated and identified. The objective of the present work was to investigate the solubility of the fractionated aggregating peptides, as a function of concentration, and their aggregating capacities toward added intact proteins. The amount of aggregated material and the composition of the aggregates obtained were measured by nitrogen concentration and size exclusion chromatography, respectively. The results showed that of the four fractions obtained from the aggregating peptides, two were insoluble, while the other two consisted of 1:1 mixture of low and high solubility peptides. Therefore, insoluble peptides coaggregated, assumedly via hydrophobic interactions, other relatively more soluble peptides. It was also shown that aggregating peptides could aggregate intact protein nonspecifically since the same peptides were involved in the aggregation of whey proteins, beta-casein, and bovine serum albumin. Both insoluble and partly insoluble peptides were required for the aggregation of intact protein. These results are of interest for the applications of protein hydrolysates, as mixtures of intact protein and peptides are often present in these applications.     

Hydrolysis of whey protein isolate with Bacillus licheniformis protease: fractionation and identification of aggregating peptides

The objective of this work was to identify the dominant aggregating peptides from a whey protein hydrolysate (degree of hydrolysis of 6.8%) obtained with Bacillus licheniformis protease. The aggregating peptides were fractionated with preparative reversed-phase chromatography and identified with liquid chromatography-mass spectrometry. The results showed that the dominant aggregating peptide, at pH 7.0, was beta-lg AB [f1-45]. In addition, the peptides beta-lg AB [f90-108]-S-S-alpha-la [f50-113], alpha-la [f12-49]-S-S-alpha-la [f50-113], beta-lg AB [f90-108]-S-S-beta-lg AB [f90-108], beta-lg A [f90-157], and beta-lg AB [f135-157/158] were also identified as main aggregating peptides. The results further showed that aggregation, via hydrophobic interactions, prevented further digestion (at pH 8.0), thereby explaining the large size of the aggregating peptides. It is hypothesized that B. licheniformis protease breaks down hydrophilic segments in the substrate and, therefore, preserves hydrophobic segments that aggregate once exposed to the solvent.     

Sorption of dissolved organic nitrogen by acid subsoil horizons and individual mineral phases

Dissolved organic matter is important in translocation and export of nutrients from forest ecosystems. Its mobility in soil is restricted by sorption to mineral surfaces which depends on its chemical properties. Carboxyl and hydroxyl groups form strong bondings to mineral surfaces, whereas the role of N‐containing functional groups in the sorption process is less well understood. We examined in laboratory experiments the binding of dissolved organic matter from the forest floor to amorphous Al(OH)₃, goethite, kaolinite, and illite and to subsoils in order to compare the sorption and desorption of dissolved organic C with that of dissolved organic N. The mineral samples were equilibrated with acidic solutions of organic matter at pH 4. In the equilibrium solutions organic C and N and their contribution to two operationally defined fractions, namely the so‐called hydrophilic and hydrophobic fractions, were determined. We measured neutral and acidic amino sugars to discover the nature of the binding of organic N. Within the hydrophilic and hydrophobic fractions, the sorption and desorption of organic C and N did not differ, indicating that there was no preferential binding of N‐containing compounds. The hydrophilic fraction contained more N and sorbed less than the hydrophobic fraction, and so the overall retention of organic N by the mineral phases and subsoils was smaller than that of organic C. Among the amino sugar compounds, muramic acid was preferentially removed from the solution, whereas the neutral amino sugars were sorbed similar to organic C. The results suggest that the sorption of N‐containing compounds is favoured by acidic groups and not by amino groups.     

Isotopic fractionation of dissolved organic carbon in shallow forest soils as affected by sorption

Isotopic fractionation of dissolved organic carbon percolating through the soil is often interpreted as due to microbial transformation. We investigated the potential effects of sorption on the δ¹³C of dissolved organic C in field and laboratory experiments. We sampled the organic C in soil water at two forested sites and measured sorption with intact mineral soil and individual minerals (dolomite, ferrihydrite, goethite, and quartz). The dissolved organic C was separated into hydrophilic and hydrophobic fractions using a resin approach. The δ¹³C values of bulk soils, alkaline‐extractable organic C, and dissolved organic C and its fractions were measured. Hydrophilic and hydrophobic fractions in forest floor seepage water were characterized by ¹³C‐NMR spectroscopy. At both sites, δ¹³C of dissolved organic C increased with increasing depth, suggesting that decomposition contributes to the loss of the dissolved organic C. However, there was an enrichment of hydrophilic organic C in the soil solution as the water moved down the soil. The δ¹³C values of hydrophilic fractions were less negative than those of hydrophobic fractions. The smaller δ¹³C in the hydrophobic fraction was due to the large contribution of compounds derived from lignin that are depleted in ¹³C. As the isotope composition of both fractions of dissolved organic C did not change throughout the profile, changes in δ¹³C of total organic C reflected changes in the relative proportions of its hydrophilic and hydrophobic fractions. The sorption experiments with minerals and soil cores gave similar results. When dissolved organic C came into contact with mineral material, the δ¹³C of that remaining in solution increased due to preferential sorption of the ¹³C‐depleted hydrophobic fractions. Moreover, the soils released hydrophilic organic C with large δ¹³C values, increasing the δ¹³C of organic C in effluents from soil compared with that in the inflow. Thus, selective sorption of organic C fractions changes δ¹³C in a way that mimics metabolic transformation and decomposition.     

Antioxidant activity of peptides obtained from porcine myofibrillar proteins by protease treatment

Hydrolysates obtained from porcine myofibrillar proteins by protease treatment (papain or actinase E) exhibited high antioxidant activity in a linolenic acid peroxidation system induced by Fe(2+). Hydrolysates produced by both papain and actinase E showed higher activities at pH 7.1 than at pH 5.4. The antioxidant activity of the papain hydrolysate was almost the same as that of vitamin E at pH 7.0. These hydrolysates possessed 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and chelating activity toward metal ions. Antioxidant peptides were separated from the papain hydrolysate by ion exchange chromatography. The acidic fraction obtained by this method exhibited higher activity than the neutral or basic fractions. Antioxidant peptides in the acidic fraction were isolated by high-performance liquid chromatography on an ODS column and shown to possess the structures DSGVT, IEAEGE, DAQEKLE, EELDNALN, and VPSIDDQEELM. The DAQEKLE peptide showed the highest activity among these peptides.     

Preparation of whey protein hydrolysates using a single- and two-stage enzymatic membrane reactor and their immunological and antioxidant properties: characterization by multivariate data analysis

An initial 5% (w/v), followed thereafter with replacement aliquots of 3% (w/v), whey protein isolate (WPI) (ca. 86.98% Kjeldahl N x 6.38), was hydrolyzed using Protease N Amano G (IUB 3.4.24.28, Bacillus subtilis) in an enzymatic membrane reactor (EMR) fitted with either a 10 or 3 kDa nominal molecular weight cutoff (NMWCO) tangential flow filter (TFF) membrane. The hydrolysates were desalted by adsorption onto a styrene-based macroporous adsorption resin (MAR) and washed with deionized water to remove the alkali, and the peptides were desorbed with 25, 50, and 95% (v/v) ethyl alcohol. The desalted hydrolysates were analyzed for antibody binding, free radical scavenging, and molecular mass analysis as well as total and free amino acids (FAA). For the first time a quantity called IC50, the concentration of peptides causing 50% inhibition of the available antibody, is introduced to quantify inhibition enzyme-linked immunosorbent assay (ELISA) properties. Principal component analysis (PCA) was used for data reduction. The hydrolysate molecular mass provided the most prominent influence (PC1 = 57.35%), followed by inhibition ELISA (PC2 = 18.90%) and the antioxidant properties (PC3 = 10.43%). Ash was significantly reduced in the desalted fractions; the protein adsorption recoveries were high, whereas desorption with alcohol was prominently influenced by the hydrophobic/ hydrophilic amino acid balance. After hydrolysis, some hydrolysates showed increased ELISA reactivity compared with the native WPI.     

Differences in the chemical composition of dissolved organic matter from waste material of different sources

The chemical composition of waste-material-derived dissolved organic matter (DOM) was characterized by chemolytic analyses and ¹H, ¹³C and ³¹P nuclear magnetic resonance (NMR) spectroscopy. Dissolved organic matter was extracted by water from an aerobic fermented urban waste compost, a sewage sludge and a pig slurry and then fractionated using the XAD-8 method. The amount of water-extractable dissolved organic carbon (DOC) ranged from 3% in the sewage sludge to 22% in the pig slurry. Dissolved organic matter isolated from pig slurry was equally distributed between hydrophilic and hydrophobic DOC, whereas in the sewage-sludge-derived material the hydrophobic fraction was predominant. Dissolved organic C from the urban waste compost was mainly within the hydrophilic fraction. Wet-chemical analysis and ¹H- and ¹³C-NMR spectra showed that both DOM fractions from the urban waste compost were low in neutral, acidic and amino sugars as well as in lignin-derived compounds. In turn, the materials were rich in low-molecular-weight aliphatic compounds. The chemical structure of both fractions is probably the result of the intensive transformation of urban waste compost during its fermentation. The hydrophilic fractions of DOM from sewage sludge and pig slurry contained considerable amounts of carbohydrates but were also rich in low-molecular-weight aliphatics. The respective hydrophobic fractions had the largest contents of CuO-extractable phenols which may in part derive from sources other than lignin. By contrast with the other materials, the hydrophobic fraction from the pig slurry seemed to contain polymeric rather than low-molecular-weight material. The ³¹P-NMR spectrum of the hydrophilic DOM fraction from urban waste compost did not show signals of inorganic or organic P compounds while the spectrum of the hydrophobic fraction revealed traces of monoester P, diester P, and orthophosphate. ³¹P-NMR spectroscopy suggested that both the hydrophobic and hydrophilic fractions from pig slurry did not contain organic P. The hydrophilic DOM fraction from sewage sludge contained orthophosphate, organic monoester P and a little pyrophosphate. The hydrophobic fraction contained mainly organic diester P and smaller amounts of teichoic acids and organic monoester P. Considering that water-soluble fractions of urban waste compost contained no easily plant-available P and a low content of labile organics, we conclude that this material contains less labile nutrients and is more refractory than the soluble constituents of pig slurry and sewage sludge.     

土壤调理剂界面改性对黄灌区盐碱离子迁移特性的影响

[目的] 通过调理剂进行亲水和疏水处理,探究调理剂不同界面特性对黄灌区土壤水盐运移规律的影响,揭示粉煤灰基土壤调理剂对盐碱地的改良机理,为不同类型盐碱地改良提供科学参考。[方法] 以宁夏回族自治区苏打盐碱土为例,设置土柱试验探究调理剂改性前后土壤中不同盐离子在空间上的运移规律。[结果] 添加未改性调理剂、亲水改性调理剂、疏水改性调理剂后,表层脱盐率分别为79.72%,59.91%和84.79%,远高于未加调理剂的12.64%。较添加未改性调理剂组相比,亲水性处理组含水量提高6.59%,全盐量提高了47.13%;疏水性处理组土壤含水量降低了0.67%,全盐量降低了25.22%;pH值分别从10.5降低至8.08,8.71;表层Na⁺含量分别降低10.39%和30.46%,K⁺含量分别降低10.22%和45.93%。[结论] 调理剂的界面特性可显著影响土壤中水盐运移。疏水处理更能提升调理剂对盐碱地的改良效果,强化盐分随土壤水的下渗对盐碱地改良更为重要。     

Contribution of soluble and insoluble fractions of agricultural residues to short‐term pH changes

The retention of agricultural residues in cropping systems to maintain soil fertility is also important for the redistribution of alkalinity. In systems that adopt minimum or no‐tillage practices residue incorporation into the soil may occur slowly and the contribution of soluble and insoluble residue fractions to pH change may vary temporally and spatially. In this study we examined the contribution of whole, water soluble (70°C for 1 hour for two cycles) and insoluble fractions of canola, chickpea and wheat residues (added at 10 g kg^?1 soil) to pH change in a Podosol (Podzol; initial pH 4.5) and a Tenosol (Cambisol; initial pH 6.2) over a 59‐day incubation period. Whole residues increased pH in both soils, with the magnitude of the pH increase (chickpea > canola > wheat) being related to alkalinity content (concentration of excess cations) of the residue. Temporal release of alkalinity was only observed for the larger alkalinity content canola and chickpea residues and the change in pH was greater than during the initial period (approximately 4 hours; T0). Increases in pH were attributed to the decarboxylation of organic anions and the association of H⁺ with organic anions and other negatively charged chemical functional groups. The relative contribution of these processes depended on the residue and the initial soil pH. Our results show that 40–62% of the alkalinity of canola and chickpea residues resided in the soluble fraction. Furthermore, pH increases caused by soluble fractions may be transient if these contain large N concentrations. Soil properties that influence inorganic N dynamics such as inhibition of nitrification at acid pH will be important in determining the subsequent direction and magnitude of pH change.     

Dissolved organic phosphorus and sulphur as influenced by sorptive interactions with mineral subsoil horizons

This study tested the hypothesis that, like dissolved organic nitrogen (N), dissolved organic phosphorus (P) and sulphur (S) are more mobile in soil than is organic carbon (C). To do so, I compared the sorption of organic P and S to subsoil materials with that of organic C. Soil samples were equilibrated with water‐soluble organic matter from the forest floor at pH 4 and in the equilibrium solutions organic C, P, and S, and their distributions between the hydrophilic and hydrophobic fraction were determined. Sorption of C within the organic matter did not differ from that of P and S. However, the hydrophilic fraction contained the vast majority of P and S and sorbed far less than the hydrophobic fraction. So the overall retention of organic P and S was smaller than that of organic C. This result suggested that dissolved organic matter is more important in the loss of plant nutrients than in the release of C from soil.     

Enzymatic hydrolysis as a means of expanding the cold gelation conditions of soy proteins

Acid-induced cold gelation of soy protein hydrolysates was studied. Hydrolysates with degrees of hydrolysis (DH) of up to 10% were prepared by using subtilisin Carlsberg. The enzyme was inhibited to uncouple the hydrolysis from the subsequent gelation; the latter was induced by the addition of glucono-delta-lactone. Visual observations, confocal scanning laser microscopy images, and the elasticity modulus showed that hydrolysates gelled at higher pH values with increasing DH. The nonhydrolyzed soy protein isolate gelled at pH approximately 6.0, whereas a DH = 5% hydrolysate gelled at pH approximately 7.6. Gels made from hydrolysates had a softer texture when manually disrupted and showed syneresis below a pH of 5-5.5. Monitoring of gelation by measuring the development of the storage modulus could be replaced by measuring the pH onset of aggregate formation (pH(Aggr-onset)) using turbidity measurements. The rate of acidification was observed to also influence this pH(Aggr-onset). Changes in ionic strength (0.03, 0.2, and 0.5 M) had only a minor influence on the pH(Aggr-onset), indicating that the aggregation is not simply a balance between repulsive electrostatic and attractive hydrophobic interactions, but is much more complex.     

Contribution of Hydrophobic Soluble Gluten Proteins,Fractionated by Hydrophobic Interaction Chromatography in Highly Acetylated Agarose,to Dough Rheological Properties

Hydrophobic interaction chromatography with highly acetylated agarose in 1‐mL columns was used to fractionate gliadins and acid‐soluble glutenins. Proteins were eluted in two fractions, the first with acetate buffer (pH 3.6) containing 35% propanol, and the second with Tris buffer in 8M urea. The proportion of eluted protein in the second fraction was called the surface hydrophobicity index. The study included 20 wheat samples of different baking qualities. Multiple regression analysis using the general linear model combined with the stepwise technique was used to relate the surface hydrophobicity index of soluble gluten proteins to specific dough rheological characteristics. Surface hydrophobicity index of gliadins and acetic acid soluble glutenins explained part of the variability of swelling index, extensibility, and work of deformation (dough strength) measured with the alveograph, and part of the farinograph water absorption variability, but showed no relationship to dough mixing characteristics. Hydrophobic soluble gluten proteins fractionated by hydrophobic interaction chromatography (HIC) explained a part of the variability of dough rheological properties.     

Water-soluble organic matter and soil solution acidity in the main soil types of the central forest state biosphere reserve

The dynamics of the soluble organic carbon (C_WSO) and pH in soil solutions of the main soil types of the Central Forest State Biosphere Reserve were described. Possible reasons for the pH variation were considered. It was found that different soils are characterized by specific levels of pH and C_WSO. The structure of the soil solution acidity and its changes with depth were analyzed, and the relationship between the acidity and the content of water-soluble organics substances (WSOs) was determined. The predominantly hydrophilic nature and polydispersity of WSOs in soil solutions were revealed, as well as a decrease in the proportion of high-molecular soil fractions with depth. It was found that the mobility of metals in soils depends on the amphiphilic properties of WSOs to which they are bound, the metal features, and the soil type. The possibility of selective absorption of metals bound to the hydrophobic WSO fraction was shown.     

Inhibition of ice crystal growth in ice cream mix by gelatin hydrolysate

The inhibition of ice crystal growth in ice cream mix by gelatin hydrolysate produced by papain action was studied. The ice crystal growth was monitored by thermal cycling between -14 and -12 degrees C at a rate of one cycle per 3 min. It is shown that the hydrolysate fraction containing peptides in the molecular weight range of about 2000-5000 Da exhibited the highest inhibitory activity on ice crystal growth in ice cream mix, whereas fractions containing peptides greater than 7000 Da did not inhibit ice crystal growth. The size distribution of gelatin peptides formed in the hydrolysate was influenced by the pH of hydrolysis. The optimum hydrolysis conditions for producing peptides with maximum ice crystal growth inhibitory activity was pH 7 at 37 degrees C for 10 min at a papain to gelatin ratio of 1:100. However, this may depend on the type and source of gelatin. The possible mechanism of ice crystal growth inhibition by peptides from gelatin is discussed. Molecular modeling of model gelatin peptides revealed that they form an oxygen triad plane at the C-terminus with oxygen-oxygen distances similar to those found in ice nuclei. Binding of this oxygen triad plane to the prism face of ice nuclei via hydrogen bonding appears to be the mechanism by which gelatin hydrolysate might be inhibiting ice crystal growth in ice cream mix.     

宜兴市温室土壤理化性质的调查和分析

对宜兴市的温室和露天土壤的理化性质(包括pH,电导率,各种水溶性离子的含量以及全盐量)进行调查测定,结果表明:宜兴市温室土壤pH值0-20 cm土层比深层低,有酸化趋势;土壤全盐量显著高于露天土壤,盐分已明显在0～20cm土层富集,表层土壤全盐含量均在2.00 g/kg以上;在盐分的离子组成中,阳离子以Ca2 为主,平均占全盐总量的14%,阴离子以NO3-和SO42-为主,分别平均占全盐总量的56%和23%.说明该地区温室土壤己达到中度盐渍化水平.     

Tea Plantation–Induced Activation of Soil Heavy Metals

Abstract

The accumulation of heavy metals in tea leaves is of concern because of its impact on tea quality. This study characterized long‐term changes of soil properties and heavy‐metal fractions in tea gardens and their effect on the uptake of metals from soils by the plants. Soil and tea leaf samples were collected from five plantations with a history of 2–70 years in Jinghua, Zhejiang Province, southeast China. The six chemical fractions (water‐soluble, exchangeable, carbonate‐bound, organic‐matterbound, oxide‐bound, and residual forms) of cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), nickel (Ni), manganese (Mn), lead (Pb), and zinc (Zn) in the soils were characterized. Dissolved organic‐matter accumulation in the soils and effects of low‐molecular‐weight organic acids on solubility of soil heavy metals were also tested. Long‐term tea plantation use resulted in accumulation of dissolved organic matter, decrease of soil pH, and elevation of water‐soluble and exchangeable metal fractions, thereby increasing metal contents in leaves. The influence was more significant when soil pH was less than 4.4. The results indicated that both acidification and accumulation of dissolved organic matter induced by tea plantations were also important causes of increased accumulation of the metals in the tea leaves. This was particularly true for the soils polluted with low concentration of heavy metals, because availability of the metals in these soils was mainly controlled by pH and dissolved organic matter.     

Transformation of organic matter from maize residues into labile and humic fractions of three European soils as revealed by 13C distribution and CPMAS-NMR spectra

The dynamics of incorporation of fresh organic residues into the various fractions of soil organic matter have yet to be clarified in terms of chemical structures and mechanisms involved. We studied by ¹³C‐dilution analysis and CPMAS‐¹³C‐NMR spectroscopy the distribution of organic carbon from mixed or mulched maize residues into specific defined fractions such as carbohydrates and humic fractions isolated by selective extractants in a year‐long incubation of three European soils. The contents of carbohydrates in soil particle size fractions and relative δ¹³C values showed no retention of carbohydrates from maize but rather decomposition of those from native organic matter in the soil. By contrast, CPMAS‐¹³C‐NMR spectra of humic (HA) and fulvic acids (FA) extracted by alkaline solution generally indicated the transfer of maize C (mostly carbohydrates and peptides) into humic materials, whereas spectra of organic matter extracted with an acetone solution (HE) indicated solubilization of an aliphatic‐rich, hydrophobic fraction that seemed not to contain any C from maize. The abundance of ¹³C showed that all humic fractions behaved as a sink for C from maize residues but the FA fraction was related to the turnover of fresh organic matter more than the HA. Removal of hydrophobic components from incubated soils by acetone solution allowed a subsequent extraction of HA and, especially, FA still containing much C from maize. The combination of isotopic measurements and NMR spectra indicated that while hydrophilic compounds from maize were retained in HA and FA, hydrophobic components in the HE fraction had chemical features similar to those of humin. Our results show that the organic compounds released in soils by mineralization of fresh plant residues are stored mainly in the hydrophilic fraction of humic substances which are, in turn, stabilized against microbial degradation by the most hydrophobic humic matter. Our findings suggest that native soil humic substances contribute to the accumulation of new organic matter in soils.