Effect of Tempering Conditions on Milling Performance and Flour Functionality

Tempering conditions of wheat grain change the quality of the flour, yet most experimental milling systems use a standard tempering without optimization. The effect of tempering condition on milling performance and flour functionality for soft red winter (SRW) wheat grain was tested by measuring flour yield, ash, polyphenol oxidase (PPO), and solvent retention capacity (SRC) in grain samples from three SRW cultivars (Roane, Cyrus, and Severn). Tempering was conducted with a full factorial design of initial wheat moisture, tempered wheat moisture, tempering temperature, and tempering time at two levels. Tempered wheat moisture had the largest effect on milling performance and flour functionality. Flour yield was more reduced for all samples tempered at 15% moisture than for samples tempered to 12% moisture. Flour quality of the 15% tempered sample was better than the 12% tempered samples due to less bran contamination as measured by flour ash and PPO. Increasing the tempering moisture increased flour sucrose SRC and lactic acid SRC but reduced sodium carbonate SRC for samples. Changing tempered wheat moisture changed flour yield and quality much more than did changing the length of time for tempering, the temperature at wheat is tempered, or differences in the initial moisture of the wheat before tempering. The last three effects could be used to improve flour yield in both the 12 and 15% tempered wheat treatment but the detrimental effects of these treatments on flour quality were minimal when combined with the 15% tempered wheat moisture treatment.     

Analysis of amino acids by gas chromatography as the N-trifluoroacetyl n-butyl esters

This presentation describes amino acid analysis with the gas chromatographic method and experimental conditions using the N-trifluoroacetyl n-butyl ester derivatives; the study we describe here was undertaken to compare gas chromatographic (GC) and ion-exchange chromatographic (IEC) analyses of amino acids in hydrolysates of 9 diverse sample types to gain insight into effects of these 2 chromatographic methods of analysis on variation in amino acid results. Our study showed that values for samples prepared by 2 separate laboratories using the same procedure were generally in good agreement when all of the hydrolysates were analyzed by a single laboratory using a single method of analysis. To compare results from gas chromatography with those from ion-exchange chromatography analyses were performed by 2 different laboratories on the same hydrolysates and on different hydrolysates prepared by the same method by both laboratories. The data demonstrate that GC and IEC can be expected to yield essentially identical results when applied to the same hydrolysate. Agreement is so close that interlaboratory differences in hydrolysate preparation of the same sample contribute as much to variation in amino acid results as does the method of analysis, a fact which should be noted in planning collaborative studies.     

Determination of free and total phenolic acids in plant-derived foods by HPLC with diode-array detection

A high-performance liquid chromatographic (HPLC) method with diode-array detection (DAD) was used to identify and quantify free and total phenolic acids (m-hydroxybenzoic acid, p-hydroxybenzoic acid, protocatechuic acid, gallic acid, vanillic acid, syringic acid, o-coumaric acid, m-coumaric acid, p-coumaric acid, caffeic acid, ferulic acid, sinapic acid, chlorogenic acid, and ellagic acid) in plant foods. Free phenolic acids were extracted with a mixture of methanol and 10% acetic acid. Bound phenolic acids were liberated using first alkaline and then acid hydrolysis followed by extraction with diethyl ether/ethyl acetate (1:1). All fractions were quantified separately by HPLC. After HPLC quantification, results of alkali and acid hydrolysates were calculated to represent total phenolic acids. Ellagic acid was quantified separately after long (20 h) acid hydrolysis. The methods developed were effective for the determination of phenolic acids in plant foods. DAD response was linear for all phenolic acids within the ranges evaluated, with correlation coefficients exceeding 0.999. Coefficients of variation for 4-8 sample replicates were consistently below 10%. Recovery tests of phenolic acids were performed for every hydrolysis condition using several samples. Recoveries were generally good (mean >90%) with the exceptions of gallic acid and, in some cases, caffeic acid samples.     

A simple and accurate method for determining wheat grain fructan content and average degree of polymerization

An improved method for the measurement of fructans in wheat grains is presented. A mild acid treatment is used for fructan hydrolysis, followed by analysis of the released glucose and fructose with high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Not only the amount of fructose set free from fructans but also the released glucose can be quantified accurately, allowing determination of the average degree of polymerization of fructans (DP(av)). Application of the mild acid treatment to different grain samples demonstrated that a correction should be made for the presence of sucrose and raffinose, but not for stachyose or higher raffinose oligosaccharides. The fructan content and DP(av) of spelt flour, wheat flour, and whole wheat flour were 0.6%, 1.2%, and 1.8% of the total weight and 4, 5, and 6, respectively. Validation experiments demonstrate that the proposed quantification method is accurate and repeatable and that also the DP(av) determination is precise.     

Contribution of Sourdough Lactobacilli,Yeast, and Cereal Enzymes to the Generation of Amino Acids in Dough Relevant for Bread Flavor

The amino acid release was determined in wheat doughs supplied with salt, acid, dithiothreitol, or starter cultures to evaluate the relevance of the amino acid concentration on bread flavor. Wheat flour proteinases almost linearly released amino acids and the highest activity of wheat flour proteinases was found in acidified and reduced doughs. The effects of starter cultures on amino acid concentrations depended on their composition. Yeasts exhibited a high demand for amino acids, however, the total amino acid concentrations were not markedly affected by lactic acid bacteria. The individual amino acid contents were determined by the pH during fermentation and microbial metabolism. The formation of proline was favored by values higher than pH 5.5, whereas release of phenylalanine, leucine and cysteine mainly occurred at lower pH. Ornithine was found only in doughs fermented with Lactobacillus pontis. To determine effects of the amino acid concentration on bread aroma, fermented doughs were evaluated in baking experiments. An increased intensity of bread flavor was obtained by preferments prepared with lactic acid bacteria. The roasty note of wheat bread crust could be markedly enhanced by L. pontis. This results support the assumption that flavor of wheat bread is enhanced by increasing the concentration of free amino acids and especially ornithine in dough.     

脱酰胺与双酶协同作用提高小麦面筋蛋白酶解效率

为了探讨了不同脱酰胺处理和双酶协同作用方式对小麦面筋蛋白酶解效率及其产物抗氧化活性的影响,该文研究了小麦面筋蛋白在各种预处理方式和酶解条件下的蛋白回收率、水解度、抗氧化性能及肽分子量分布情况。结果显示,单独热处理(90℃,30 min)小麦面筋蛋白对其酶解效率无显著影响,而采用添加0.5 mol/L柠檬酸溶液进行热处理(质量分数为5%,90℃,30 min)可显著(P0.05)提高其蛋白回收率。此外,酶制剂添加顺序及双酶共同水解作用时间对酶解效率均具有较大影响:加入谷氨酰胺酶预先水解对小麦面筋蛋白的深度水解有促进作用;一定时间内的双酶协同作用有利于酶解的进行,但较长时间的双酶作用反而会抑制酶解效率。采用谷氨酰胺酶(质量分数为0.2%)对经柠檬酸加热处理的小麦面筋蛋白作用12 h后再加入胰酶(质量分数为0.6%)共同作用7 h可使蛋白回收率达70.74%,水解度达到9.88%;另外,酶解产物的自由基清除能力ABTS+(2,2’-Azinobis-(3-ethylbenzthiazoline-6-sulphonate)+)值与氧化自由基吸收能力(ORAC,oxygen radical absorbance capacity)值分别达到478.95 mmol/g和213.85μmol/g,提示该酶解产物是一种潜在优秀食品抗氧化剂。研究结果可为拓宽小麦面筋蛋白的应用领域,以及高效制备抗氧化活性肽提供方法和理论指导。     

Environmentally induced changes in amino acid composition in the grain of durum wheat grown under different water and temperature regimes in a Mediterranean environment

Amino acid composition is an important feature in determining the nutritional value of wheat grain for human and animal diets. Environmental conditions are known to influence protein quantity as well as grain production and, in turn, amino acid composition. In this study, grain yield, protein content, and amino acid composition were determined in 10 durum wheat genotypes under three water and temperature regimes in a Mediterranean environment. The highest value for grain-protein content (15.7%) was found in the warmer and driest environment and the lowest (12.8%) in the irrigated environment. Although amino acid composition showed significant variation for all genotypes, with the exception of arginine and cysteine, major changes in amino acid composition were caused by environmental conditions and in particular by water availability and temperature during the grain-filling period, which significantly altered the duration of grain development. The amino acids with the highest percentage of variation between environments were tyrosine (26.4%), lysine (23.7%), methionine (20.3%), threonine (19.3%), and valine (15.6%), whereas phenylalanine (5.1%), glycine (6.4%), and aspartic acid (6.8%) showed the least variation between environments. Whereas the content of glutamine, phenylalanine, and proline increased with the decrease in grain-filling duration, the remaining amino acids tended to diminish, presumably because high temperature and drought favored the deposition of gliadins (proteins particularly rich in glutamine and proline), to the detriment of albumins and globulins (proteins especially rich in threonine, lysine, methionine, valine, and histidine). Despite the negative correlations found between the percentage of protein and its content in essential amino acids, the results indicate that reductions in lysine per unit of food were not very pronounced (0.32 to 0.29 g/100 g of flour) with increases of up 22.7% in grain-protein content, whereas threonine did not change and valine even slightly increased.     

Contents of phenolic acids, alkyl- and alkenylresorcinols, and avenanthramides in commercial grain products

The contents of free and total phenolic acids and alk(en)ylresorcinols were analyzed in commercial products of eight grains: oat (Avena sativa), wheat (Triticum spp.), rye (Secale cerale), barley (Hordeum vulgare), buckwheat (Fagopyrum esculentum), millet (Panicum miliaceum), rice (Oryza sativa), and corn (Zea mays). Avenanthramides were determined in three oat products. Free phenolic acids, alk(en)ylresorcinols, and avenanthramides were extracted with methanolic acetic acid, 100% methanol, and 80% methanol, respectively, and quantified by HPLC. The contents of total phenolic acids were quantified by HPLC analysis after alkaline and acid hydrolyses. The highest contents of total phenolic acids were in brans of wheat (4527 mg/kg) and rye (4190 mg/kg) and in whole-grain flours of these grains (1342 and 1366 mg/kg, respectively). In other products, the contents varied from 111 mg/kg (white wheat bread) to 765 mg/kg (whole-grain rye bread). Common phenolic acids found in the grain products were ferulic acid (most abundant), ferulic acid dehydrodimers, sinapic acid, and p-coumaric acid. The grain products were found to contain either none or only low amounts of free phenolic acids. The content of avenanthramides in oat flakes (26-27 mg/kg) was about double that found in oat bran (13 mg/kg). The highest contents of alk(en)ylresorcinols were observed in brans of rye (4108 mg/kg) and wheat (3225 mg/kg). In addition, whole-grain rye products (rye bread, rye flour, and whole-wheat flour) contained considerable levels of alk(en)ylresorcinols (524, 927, and 759 mg/kg, respectively).     

Simple Phenolic Acids in Flours Prepared from Canadian Wheat: Relationship to Ash Content,Color, and Polyphenol Oxidase Activity

Simple phenolic acid levels were determined on pooled millstreams of five different classes of Canadian wheat milled to ~75, 80, and 85% extraction. Pooled flours and whole grain were analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC) to establish endogenous levels of insoluble bound, soluble esterified, and free phenolic acids. Only ferulic acid was detected in the insoluble bound category, which accounted for >80% of the total phenolic acids present in every flour. The soluble esterified phenolic acids accounted for up to 17% of the overall total phenolic acid content within a flour. The major constituents were sinapic, ferulic, and vanillic acids, with minor amounts of coumaric, caffeic, and syringic acids. Free phenolic acids accounted for a maximum of 6% of the total phenolic content of any prepared flour. Ferulic acid was the major free phenolic acid, while sinapic acid was not detected in any flour. Significant correlations (r = 0.64–0.97, P < 0.05) were observed between insoluble bound ferulic acid, individual soluble esterified acids, and most free acids with polyphenol oxidase activity, as well as color and ash content for each class.     

High levels of sugars and fructan in mature seed of sweet wheat lacking GBSSI and SSIIa enzymes

Sweet wheat (SW), which lacks functional granule-bound starch synthase I (GBSSI) and starch synthase IIa (SSIIa), accumulates high levels of free sugars in immature seeds. Here, we examined the effects of the lack of these two enzymes on mature kernel composition. Whole grain flour of SW had higher levels of sugars, particularly maltose, slightly higher ash and protein content, approximately two to three times higher lipid levels, and about twice as much total dietary fiber as parental or wild-type lines. Considerably higher levels of low-molecular-weight soluble dietary fiber (LMW-SDF), largely consisting of fructan, were also detected in SW. Although there were no differences in total amino acid levels, the free amino acid content of SW was approximately 4-fold higher than that of wild type, and the levels of certain free amino acids such as proline were particularly high. Thus, we were able to clearly demonstrate that the lack of GBSSI and SSIIa caused dramatic changes in mature seed composition in SW. These compositional changes suggest that SW flour may provide health benefits when used as a food ingredient.     

Influence and Interactions of Processing Conditions and Starter Culture on Formation of Acids,Volatile Compounds,and Amino Acids in Wheat Sourdoughs

The aim of this work was to study the influence of process parameters and the starter culture on the characteristics of wheat sourdough by using response surface methodology. Influence of fermentation temperature (16–32°C), ash content of flour (0.6–1.8%), and fermentation time (6–20 hr) were considered as independent factors and their effects were studied in sourdough fermented with Lactobacillus plantarum, L. brevis, Saccharomyces cerevisiae, or with a combination of yeast and lactic acid bacteria. Formation of acidity, free amino acids, and volatile compounds were considered the main responses. A possibility to enhance formation of potential flavor compounds and precursors without excessive acidity formation in wheat sourdoughs was established. The total amount of amino acids increased by 25–50%, depending on the strain and fermentation conditions. The total amount of volatile compounds increased seven‐ to 100‐fold, depending on the strain and fermentation conditions. Sourdough started with S. cerevisiae was an effective way to optimize the amount of volatile compounds without excessive acidity formation in appropriate processing conditions. Ash content of flour and fermentation time were the most significant factors to modify metabolic activity of wheat sourdoughs. Frequent interactions between the studied factors were observed on the formation of acidity, amino acids, and volatile compounds with most of the strains studied. Possibility to improve current industrial fermentation processes and control flavor attributes of breads by using optimized sourdough was established.     

Variation in Polyphenol Oxidase Activity and Quality Characteristics Among Hard White Wheat and Hard Red Winter Wheat Samples

Polyphenol oxidase (PPO) has been related to an undesirable brown discoloration of wheat-based end products. Consumer acceptance and product quality are generally decreased by the darkening phenomena. Two sets of wheat samples (Triticum aestivum L.) were investigated for variation in grain and flour PPO levels. Samples included 40 advanced experimental hard white winter wheat lines grown at two Kansas locations and 10 hard red winter wheat genotypes grown at three Nebraska locations. The variability in grain and flour PPO activities was influenced by growing location and population for the hard white wheat samples. There also was a significant influence of population by growing location interactions on PPO activity in both grain and flour. Genotype and growing location both contributed to variability in flour PPO activity among the hard red wheat samples. The variation in flour PPO activities among growing locations appeared larger than variation produced by genotypes tested for the hard red wheat samples. Quality parameters, such as wheat physical properties, flour protein and ash contents, grain color, and milling yield significantly correlated with grain and flour PPO activities. Among red wheat samples, flour PPO activity was related to 100 kernel weight, first reduction flour yield, and flour ash content. Grain PPO activity was related to variation in grain color observed among hard white samples. The relationship of quality characteristics with grain and flour PPO activities varied among white and red wheat samples.     

Irradiation stability of folic Acid in powder and aqueous solution

This study attempts to examine the folic acid stability after irradiation treatment, under different physical states, pH values, and atmosphere conditions. Aqueous folic acid samples, folic acid in powder, and wheat flour fortified with folic acid were irradiated by an electron beam (E-beam) between 0 (control) and 10.0 kGy. It was realized that the physical state of folic acid plays an important role on its stability toward E-beam processing, being largely unstable in solution, no matter the pH and atmosphere conditions assayed. Otherwise, folic acid in powder showed huge irradiation stability, even when mixed in a dry food matrix, such as fortified wheat flour samples.     

Phenolic acids in wheat varieties in the HEALTHGRAIN Diversity Screen

The amounts and compositions of free, conjugated, bound, and total phenolic acids were determined in 175 samples of wheat flour grown on a single site in 2005. The highest contents of total phenolic acids were found in flours of winter wheat (1171 microg/g) with average levels of 658 microg/g total phenolics across all of the wheat genotypes. Winter wheats showed a range of >3.5-fold across the concentration range for total phenolic acids. Spelt genotypes displayed the narrowest (1.9-fold) range of total phenolic acid concentration. The concentrations of phenolic acids in the different phenolic acid fractions were in the order bound > conjugated > free, with bound phenolic acids making up around 77% of the total phenolic acid concentration and free phenolic acids constituting between 0.5 and 1%. The results indicate that there is genetic diversity in phenolic acid content and that it should be possible to selectively breed for lines with high contents of phenolic components.     

Plant Sterols in Cereals and Cereal Products

The total plant sterol contents (free sterols and covalently bound structures) of the main cereals cultivated in Finland were determined. Furthermore, sterol contents were determined for different flour and bran fractions in the milling process of wheat and rye, as well as plant sterol contents in various milling and retail bakery products. The sample preparation procedure included acid and alkaline hydrolysis to liberate sterols from their glycosides and esters, respectively. Free sterols were extracted and, after recovery using solid‐phase extraction, derivatized to trimethylsilyl ethers for gas chromatography (GC) analysis. We used GC with a mass spectrometer (MS) for identification. When two cultivars of rye, wheat, barley, and oats grown in the same year were compared, the highest plant sterol content was observed in rye (mean content 95.5 mg/100 g, wb), whereas the total sterol contents (mg/100 g, wb) of wheat, barley, and oats were 69.0, 76.1, and 44.7, respectively. In addition, the 10 rye cultivars and breeding lines compared had total sterol contents of 70.7–85.6 mg/100 g. In the milling process of rye and wheat, the plant sterols fractionated according to the ash content of the corresponding milling product. In all cereal grain and milling product samples, sitosterol was the main sterol. The level of stanols differed in the different milling process samples; it was lower in the most refined rye and wheat flours (≈15%) than in the bran fractions (≈30% in the bran with 4% ash content). Rye bread with whole meal rye flour as the main or only ingredient was a good source of sterols. Sterol content was higher than that of wheat bread, whereas plant sterol content of other bakery products was affected by the type and amount of fat used in baking.     

Enzymatic solubilization of proteins in brewer's spent grain

Brewer's spent grain (BSG) is an abundant, protein-rich coproduct from the beer industry. There is a growing interest in increasing and diversifying the exploitation of BSG and related coproducts for economic and environmental reasons. In this paper, we report on a study of the solubilization of proteinaceous material from BSG using several commercial peptidase preparations. Our data show that Alcalase is the most effective peptidase for solubilization of BSG proteins, with an ability to release up to 77% of total protein. The peptides produced by Alcalase had lower average molecular weight than peptides produced by the less effective enzymes. Processes that combined peptidase treatment with carbohydrate-degrading enzyme preparations such as Depol740 increased the solubilization of dry matter (from 30 to 43% under optimal conditions). However, such additional treatment had little effect on the solubilization of protein. The choice of enzyme dosage depends on the desired hydrolysis time and was assessed through several experiments. Protein solubilization was consistently better at pH 8.0 as compared to pH 6.8. Maximum protein solubilization at pH 8.0 within 4 h required the use of 10-20 microL Alcalase per g of dry matter. However, a considerable degree of solubilization (64%) and hydrolysates with high protein content could be obtained using doses down to only 1.2 microL. Amino acid composition analyses showed that Alcalase treatment solubilizes proline and glutamine (constituents of barley hordein) slightly more efficiently than the other amino acids in BSG.     

Detection of Fragments from Internal Insects in Wheat Samples Using a Laboratory Entoleter

A simple, rapid method that uses a small mechanical rotary device (entoleter) was developed for estimating insect fragment counts in flour caused by hidden, internal‐feeding insects in whole grains of hard red winter and soft red winter wheat. Known counts of preemergent adults, pupae, and larvae of lesser grain borers and rice weevils were blended with 500 g samples of uninfested wheat. The entoleter impeller speed was adjusted based on grain hardness and moisture content to obtain about ≈98% intact and ≈2–2.5% broken kernels in an uninfested sample. The entoleter flung the wheat kernels against a surrounding steel ring. Approximately 70–90% of the insect‐infested kernels, being weaker, released internal insect pieces upon impact. The broken kernels were sieved with number 10 and number 20 sieves to obtain large‐sieved and small‐sieved fractions, respectively. Insect pieces in sieved fractions were counted. The insect piece counts were correlated with the estimated flour fragments (R² = 0.94). The entoleter method can distinguish samples of grain containing 0, 25, or 75 fragments in 50 g of flour, with greater than 95% confidence. The method can be performed in approximately 5 min per 500 g sample and could potentially be a cost‐effective method that grain handlers can use to inspect wheat loads for detecting insect damage and estimating insect fragments in flour.     

Speciation analysis of mercury in cereals by liquid chromatography chemical vapor generation inductively coupled plasma-mass spectrometry

A simple and rapid procedure for the separation and determination of inorganic, methyl, and ethyl mercury compounds was described using liquid chromatography (LC) followed by vapor generation inductively coupled plasma-mass spectrometry (VG-ICP-MS). Well resolved chromatograms were obtained within 5 min by reversed-phase liquid chromatography with a C8 column as the stationary phase and a pH 4.7 solution containing 0.5% v/v 2-mercaptoethanol and 5% v/v methanol as the mobile phase. The separated mercury compounds were converted to mercury vapors by an in situ nebulizer/vapor generation system for their introduction into ICP. The concentrations of NaBH4 and HNO3 required for vapor generation were also optimized. The method was applied for the speciation of mercury in reference materials NIST SRM 1568a Rice Flour and NIST SRM 1567a Wheat Flour and also rice flour and wheat flour samples purchased locally. The accuracy of the procedure was verified by analyzing the certified reference material NRCC DOLT-3 Dogfish Liver for methyl mercury. Precision between sample replicates was better than 13% for all the determinations. The detection limits of the mercury compounds studied were in the range 0.003-0.006 ng Hg mL(-1) in the injected solutions, which correspond to 0.02-0.06 ng g(-1) in original flour samples. A microwave-assisted extraction procedure was adopted for the extraction of mercury compounds from rice flour, wheat flour, and fish samples using a mobile phase solution.     

Characteristics and composition of watermelon, pumpkin, and paprika seed oils and flours

The nutritional quality and functional properties of paprika seed flour and seed kernel flours of pumpkin and watermelon were studied, as were the characteristics and structure of their seed oils. Paprika seed and seed kernels of pumpkin and watermelon were rich in oil and protein. All flour samples contained considerable amounts of P, K, Mg, Mn, and Ca. Paprika seed flour was superior to watermelon and pumpkin seed kernel flours in content of lysine and total essential amino acids. Oil samples had high amounts of unsaturated fatty acids with linoleic and oleic acids as the major acids. All oil samples fractionated into seven classes including triglycerides as a major lipid class. Data obtained for the oils' characteristics compare well with those of other edible oils. Antinutritional compounds such as stachyose, raffinose, verbascose, trypsin inhibitor, phytic acid, and tannins were detected in all flours. Pumpkin seed kernel flour had higher values of chemical score, essential amino acid index, and in vitro protein digestibility than the other flours examined. The first limiting amino acid was lysine for both watermelon and pumpkin seed kernel flours, but it was leucine in paprika seed flour. Protein solubility index, water and fat absorption capacities, emulsification properties, and foam stability were excellent in watermelon and pumpkin seed kernel flours and fairly good in paprika seed flour. Flour samples could be potentially added to food systems such as bakery products and ground meat formulations not only as a nutrient supplement but also as a functional agent in these formulations.     

Hydrolysis of low-molecular-weight soil peptides by immobilized proteases in column and batch reaction systems

Summary Organic matter was extracted from three soils, a cultivated Berwick sandy loam, a cultivated Franklin loamy sand, and an uncultivated Cumberland silty loam. Gel-permeation chromatography was used to separate organic matter extracts into high- (HMW) and low-molecular-weight (LMW) fractions. Reversed-phase high performance liquid chromatography was used to separate and collect the LMW peptide fractions. Peptide samples were hydrolyzed with immobilized proteases attached to beaded agarose and carboxymethyl cellulose in column and batch reaction systems. The chromatograms suggested that peptides are bound to common soil components. The amino acids released in the greatest percentages were relatively non-polar. Large percentages of serine, glycine, alanine, threonine, and valine were observed in the LMW soil peptides. Little aspartic acid, asparagine, glutamic acid, glutamine, arginine, and no histidine was detected in the LMW soil peptides. The soil peptides released different amino acid percentages and quantities when hydrolyzed by immobilized proteases attached to different supports. The quanitities of amino acids released by batch hydrolysis differed from those obtained with column hydrolysis. Greater quantities of amino acids were released (by both types of immobilized protease) from the LMW peptide hydrolysates of the two cultivated soils than from the uncultivated soil.