Extraction of amphoteric amino acids by an electromembrane process. pH and electrical state control by electrodialysis with bipolar membranes

Electrodialysis (ED) with bipolar membranes (BPM) was applied for the extraction of amino acids in an electrically neutral form. Under voltage, BPM split water, generating hydroxyl ions at the anodic side and protons at the cathodic side. The use of these membranes represents a newer method to convert neutral species into electrically charged species that are able to cross homopolar ED membranes. The ED operations were performed using a laboratory cell with a membrane area of 50 cm². A mixture containing five amino acids (Gly, Ser, Ala, Val, Phe) was examined. The concentration of each amino acid was 100 mmol dm⁻³. The current density and pH in the feed and receiver compartments were varied. Extraction degrees and current efficiencies were calculated and compared under different experimental conditions. © 1998 Society of Chemical Industry     

Extraction and characterization of soluble protein fractions from Phaseolus lunatus L seeds

Legume proteins as a potential source of valuable nutrients, are the object of several studies in order to obtain the best use. A basic knowledge becomes more important for those proteins from species not wholly utilized, before using them as food ingredients. The objective of this work was to determine several structural and nutritional characteristics of the protein fractions from Phaseolus lunatus, separated in different solvents. The relative amount of extraction for the albumins (ALB), globulins (GLB), prolamines (PRL), and glutelins (GLT) was 62.3, 34.8, 1.4 and 1.5%, respectively. The SDS-PAGE electrophoretic profile of both ALB and GLB, showed seven common bands in intervals from 10 to 95 kDa, and 14 to 99 kDa, respectively; the amino acids profile showed that PRL was the rich fraction in sulfurated amino acids (11.5 g/100 g protein); the content of lysine in the fraction of ALB was smaller than expected but the requirement of the FAO in the fractions of GLB and GLT was covered. In general, the fraction of GLB had the best balance of amino acids and digestibility (80%); however, it had a relationship of calculated protein efficiency ratio (C-PER) of 0.11, smaller than the ratio in ALB (0.97). The calorimetric analysis showed denatured temperatures around 90 degrees C for the ALB, GLB, and GLU fractions. The PRL fraction probably did not present a thermal transition because the proteins were denaturalized by the extraction conditions.     

Composition of several types of Safflower seed

The residue remaining after commercial extraction of oil from safflower seed has a greater potential as a source of animal feed or human diet supplement than is presently being realized. Safflower seed hull, kernel, and meal were analyzed to provide more information regarding their nutritive possibilities. Commercial and experimental normal hull varieties and experimental thin hull and striped hull varieties were hand separated into hull and kernel fractions and both fractions analyzed for protein, fat, fiber, ash, and amino acids. Samples of partially decorticated commercial meal and undecorticated meal, hulls, and defatted kernel from striped hull seeds were analyzed for protein, fat, fiber, ash, lignin, pentosans, anhydrouronic acid, total and reducing sugars, and amino acids. Cellulose was calculated by difference. A new factor for converting nitrogen to protein for summative analyses of safflower seed was calculated. These analyses indicate that about 15% of the nonfiber, nonash, nonprotein part of the defatted safflower kernel is of unknown composition. W. Utiliz. Res. Dev. Div., ARS, USDA.     

Heterocyclic quinones. VIII. Synthesis and spectra of new carbazoloquinone derivatives of naturally occurring amino acids

Fifteen naturally occurring amino acids representing basic, neutral and acidic members were interacted with 4,4′-dimethoxydiquinone (I). Basic amino acids reacted readily to give amino acid substituted carbazoloquinones (III), while neutral and acidic amino acids reacted only after addition of an acid binding agent. Using an excess of any of the amino acids resulted in the substitution of the quinonoid methoxyl group in compounds (III) by an amino acid moiety. In the case of basic amino acids, this substitution reaction was preferentially influenced by a terminal amino or guanidino amino rather than by an α-amino group. Moreover, basic amino acids could be reacted with two moles of (I) to produce dicarbazoloquinones (X). The i.r. and u.v.-visible spectra of the various carbazoloquinone products were determined and discussed.     

酶-膜法提取纯化荔枝核中氨基酸

采用酶-膜法提取纯化荔枝核中氨基酸。使用X JT9503中性蛋白酶酶解辅助提取荔枝核中氨基酸,实验确定酶解的优化条件为:酶用量600 U/g、酶解温度50℃、pH=6.5、酶解时间90 m in,氨基酸得率为1.16%。采用水提法和酸解提取法氨基酸得率分别仅为0.41%和0.92%。实验表明,酶解提取法要优于水提法和酸解提取法。使用截留相对分子质量50 000的陶瓷超滤膜纯化酶解提取液,实验表明,膜通量随操作压力和料液温度升高而增加。在操作压力0.22 MPa,料液温度30℃的条件下,膜的平均通量为75.63 L/(m2.h),氨基酸的截留率仅为10.3%,蛋白质的截留率为98.1%,多糖的截留率为85.2%,氨基酸能够与蛋白质、多糖等大分子实现有效分离。     

Die antioxidative Wirkung von Produkten der Maillard-Reaktion in Modellsystemen und gerösteten Haselnüssen

The Antioxidative Effect of Maillard Reaction Products in Model Systems and Roasted Hazelnuts Solutions of glucose and various amino acids were heated in phosphate buffer pH 7.0. Browning and reducing power of these Maillard reaction solutions showed correlation and increase in the ordeaicid -neutral — basic amino acids. Upon addition to emulsions of methyl imoleate (ML) in water the antioxidative effect increased in the order basic — acid — neutral amino acids. The respective amino acids alone had no effect, various Amadori compounds (fructose-alanine. -arginine and -histidine) a prooxidative effect. Extracts from roasted, defatted hazelnuts also showed correlation between browning and reducing power. Water extracts possessed higher browning and reducing power than the corresponding methanol extracts; the antioxidative effect of both extracts in ML emulsions however was comparable and increased with the roasting time as well as with the amount of extract added. The extracts were separated by gel permeation chromatography on Sephadex G-15 into three main fractions. An antioxidative effect was mainly found in the fraction of low molecular colourless Maillard reaction products.     

Chemical fractionation of shrimp extracts inducing bottom food search behavior in cod (Gadus morhua L.)

The bottom food search (BFS) feeding behavior in cod (Gadus morhua L.), has been used in a bioassay for chemical isolation of the feeding stimulant substances present in shrimp (Pandalus borealis). An aqueous methanol extract of ground shrimp was separated into acidic, neutral, and amphoteric/basic fractions by ion-exchange chromatography and into single components by preparative high-pressure liquid chromatography (HPLC). Of the isolated single components, the amino acid glycine was most potent, followed by alanine. Two unidentified substances were also highly potent. There was a synergistic effect between glycine, alanine, proline, and arginine. These four amino acids were more potent than the total amino acid pool found in the shrimp extract, indicating that there may be amino acids in this pool having an antagonistic effect.     

Fatty acid composition of rat hearts as influenced by age and dietary fatty acids

Fatty acid composition of neutral and polar lipid fractions from rat hearts was determined in rats of different ages as their diet source changed. Piebald rats were weaned at 21 days and were fed standard lab chow. Lipids from rat hearts, mothers milk and lab chow were purified on a Sephadex G-25 fine column and separated into neutral and polar lipid fractions by silicic acid column chromatography. These lipid fractions were then hydrolyzed and methylated with BF₃ in methanol, prior to gas liquid chromatographic separation on a 1/8 in. × 10 ft aluminum column of 15% EGS on 80–100 mesh acid-washed Chromosorb W. Three major fatty acids in the neutral lipid fraction comprised 72% of total neutral lipid fatty acids from young hearts. At sexual maturity (at least 74 days old) C_18∶1 was the major fatty acid, followed by C_16∶0 and C_18∶0. The same three fatty acids comprised 83% of total polar lipid fatty acids, but C_18∶0 was the major fatty acid, followed by C_16∶0 and C_18∶1. The fatty acid composition of dietary lipids influenced the total neutral lipid fatty acid composition of the rat heart, but had little influence on the fatty acid composition of the polar lipid fraction. Presented in part at the AOCS Meeting, New Orleans, April 1970.     

Preparation and nutritional characteristics of a hydrolysate from pepitona (Arca zebra)

Two soluble products resulting from the hydrolysis of pepitona (Arca zebra) were prepared as flour. Papain at its optimum hydrolysis conditions, previously established, was the enzyme used (40 degrees C for two hours at a pH of 7 in the proportion of 0.3% weight/enzyme/100 g meat). The hydrolysate obtained was then subjected to two different dehydration techniques: drum drying at 121 degrees C and 18 seconds retention, and spray drying at 101 degrees C and 40 psi pressure. The products were then stored for a five-month period at a temperature of 25 degrees C +/- 2 degrees C, time during which chemical determinations were performed in both hydrolysates. Findings showed that the time of storage does exert a significant effect of deterioration on the products. The greater and more significant quality losses occur during the first two months. The dehydration techniques used also affect significantly the soluble nitrogen content, and non-protein nitrogen and soluble solids content, as well as color of pepitona hydrolysates. Spray-drying dehydration technique does not have a significant deteriorating effect. Biological studies undertaken demonstrated that the quality of both hydrolysates is satisfactory from the nutritional and amino acid composition points of view. A protein efficiency ratio (PER) of 2.27 and 2.29 was determined for the hydrolysate dehydrated by drum drier and for the dehydrated by spray drier, respectively. With regard to amino acid composition, both had satisfactory levels of essential amino acids, with a lysine content of 6.9 g/100 g protein for the hydrolysate dehydrated by drum drying, and 8.6 g/100 g protein for the other hydrolysate dehydrated by spray drying.     

Effects of extraction conditions and alkali type on yield and composition of wheat straw hemicellulose

The utilization of various alkaline regimes for the optimal extraction and isolation of hemicellulose and cellulose from wheat straw was extensively examined. Factors in-vestigated include varying concentrations of one alkali (KOH) and H₃BO₃, varying temperature and time of extraction, and varying the nature of the alkali: calcium hy-droxide, sodium hydroxide, lithium hydroxide, and liquid ammonia were examined in this context. For example, a preferred extraction of hemicellulose from holocellulose preparations utilized a solution of 24% KOH/2% H₃BO₃ at 20°C for 2 h. This produced yields for hemicellulose and cellulose of 34.23 and 35.96%, respectively. The neutral sugar composition of the various hemicellulose fractions was found to vary slightly with treatment regime. In all hydrolysates of hemicellulose preparations, xylosé was by far the predominant sugar, comprising around 80% of the material. Minor constituents were arabinose, galactose, glucose, and uronic acids. The composition of phenolic acids and aldehydes in extracted wheat straw hemicellulose was also studied. The average molecular weights of the hemicellulose isolates ranged from 12,000 for the 30% KOH/2% H₃BO₃ (20°C, 2 h) extract to 27,000 for the extract obtained using 5% KOH/2% H₃BO₃ (20°C, 2 h). © 1996 John Wiley & Sons, Inc.     

Emulsifying and foaming properties of native and chemically modified peptides from the 2S and 12S proteins of rapeseed (Brassica napus L.)

The 2S and 12S proteins of rapeseed were isolated and subsequently hydrolyzed by pepsin or a combination of pepsin plus trypsin. The resulting hydrolysates had a 15% degree of hydrolysis and were purified by gel filtration chromatography in order to obtain homogeneous peptide fractions. Three major fractions, having an average peptide chain length of 7.5–11 amino acids, were recovered. Purified peptide fractions were acylated with butyric anhydride and sulfamidated with p-toluenesulfonyl chloride. The degree of modification was always higher than 90%. Emulsifying and foaming properties of native and chemically modified peptides were studied and compared to those of sodium dodecyl sulfate (SDS) as standard. A peptide fraction from the 15% hydrolysis of the 12S protein exhibited the best foaming properties. After sulfamidation, this peptide fraction showed a foam formation similar to that of SDS. Whereas the attachment of toluene groups generally improved the surface properties, the incorporation of an aliphatic chain of four atoms of carbon was detrimental in most of the cases. On the other hand, none of the native or hydrophobized peptide fractions was able to form a stable emulsion.     

Characterization of purified phospholipids from krill (Euphausia superba) residues deoiled by supercritical carbon dioxide

Purification of phospholipids (PL) from the Antarctic krill (Euphausia superba) using a two-step extraction process has been investigated. Using supercritical carbon dioxide (SC-CO₂) extraction with optimal extractions conditions of 45 °C, 25MPa, and CO₂ flow rate of 22 g/min, most of the neutral lipids were extracted. PC, PE and PI were then extracted in a second step conducted with modified existing method using ethanol, hexane and acetone as solvents. The major PL of krill residues was quantified by high performance liquid chromatography (HPLC-ELSD). The fatty acid compositions of total PL, PC, PE and PI were analyzed by gas chromatography (GC). A significant amount of polyunsaturated fatty acids (PUFA) was present in both total and PLs fractions. The purified PLs were characterized by their acid value, peroxide value, and the oxidative stability. The purity of PL ranged between 93 and 97% and was evaluated by spectrophotometry.     

Removal of naphthenic acids from a vacuum fraction oil with an ammonia solution of ethylene glycol

A new method to remove and purify the naphthenic acids in heavy fractions of petroleum is studied in this paper. An ammonia solution of ethylene glycol was used as the acid removal reagent by mixing with the petroleum fraction and then allowing the two phases to separate, with the naphthenic acids being extracted from petroleum fractions. The naphthenic acids were recovered by heating the ammonia solution containing naphthenic acids to release NH₃ and decompose the naphthenic acid ammonia salt. Petroleum ether was used to purify the naphthenic acids by extracting the neutral oils from the acid removal reagent. Data indicated that the optimal extraction temperature was in the range of 50–60 °C and the optimal NH₃ content in ethylene glycol was 3–5%. The contact time should be more than 10 min with the reagent/oil ratio being more than 0.3 (wt/wt). Acid removal can be greater than 85%. After purification by petroleum ether, the purity of naphthenic acids can be greater than 90%.     

Acid-Alkali Extraction of Triterpene Acids from Poria and Preparative Separation by High-Speed Counter-Current Chromatography

A method for the acid-alkali extraction and preparative separation of triterpene acids from poria was established. The triterpene acids were enriched and separated into two fractions after extraction at the optimized pH value. The two fractions were subjected to high-speed counter-current chromatography for the preparative separation of triterpene acids, separately. As a result, dehydropachymic acid, pachymic acid, 3-epi-dehydropachymic acid, poricoic acid B, dehydrotumulosic acid, and 3-epi-dehydrotumulosic acid were obtained with purities of 94.1%, 96.2%, 93.5%, 85.9%, 80.1%, and 93.1%, respectively. The structures were identified by ESI-MS, ¹H NMR, and ¹³C NMR.     

Improved Extraction of Saturated Fatty Acids but not Omega-3 Fatty Acids from Sheep Red Blood Cells Using a One-Step Extraction Procedure

Several methods are available to extract total lipid and methylate fatty acids from a range of samples including red blood cells (RBC). Fatty acid analysis of human RBC can be undertaken using a two-step extraction and methylation or a combined one-step extraction and methylation procedure. The lipid composition of sheep RBC differs significantly from that of humans and may affect their extraction. The purpose of the current study was to examine the efficiency of extraction of lipid and detection of fatty acids from sheep RBC using a one-step procedure. Fatty acids were analysed using a one-step extraction and methylation procedure using methanol:toluene and acetyl chloride in comparison with a two-step procedure involving extraction of lipid using chloroform:methanol and separate methylation. Concentrations of saturated fatty acids including C16:0 and C18:0 were significantly higher (42.6 and 33.9 % respectively) following extraction using the one-step procedure compared with the two-step procedure. However, concentrations of some polyunsaturated fatty acids, including C20:5n-3 and C22:6n-3 were not significantly different between either procedure. The improved detection of fatty acids may be related to the ability of different solvents to extract different lipid fractions. The differential extraction of lipids and detection of fatty acids from sheep RBC may have important implications in studies examining the effect of dietary treatment on the possible health benefits of fatty acids.     

猪牛鸡肉酶解液中游离氨基酸及呈味特性的对比分析

为对比分析猪肉、牛肉与鸡肉酶解液中游离氨基酸的组成及含量,采用氨基酸自动分析仪分析检测猪肉、牛肉与鸡肉酶解液中的游离氨基酸,通过计算味道强度值(TAV)确定各游离氨基酸对猪肉、牛肉与鸡肉酶解液滋味的贡献率。 结果表明,鸡肉酶解液中鲜味和甜味氨基酸质量分数以及味道强度值均大于猪肉和牛肉酶解液,3种酶解液中味道强度值最大的均为组氨酸,其次为苯丙氨酸,均为呈苦味氨基酸,可见猪、牛、鸡肉酶解液整体滋味以苦味为主。     

Production of defatted palm kernel cake protein hydrolysate as a valuable source of natural antioxidants

The aim of this study was to produce a valuable protein hydrolysate from palm kernel cake (PKC) for the development of natural antioxidants. Extracted PKC protein was hydrolyzed using different proteases (alcalase, chymotrypsin, papain, pepsin, trypsin, flavourzyme, and bromelain). Subsequently, antioxidant activity and degree of hydrolysis (DH) of each hydrolysate were evaluated using DPPH• radical scavenging activity and O-phthaldialdehyde spectrophotometric assay, respectively. The results revealed a strong correlation between DH and radical scavenging activity of the hydrolysates, where among these, protein hydrolysates produced by papain after 38 h hydrolysis exhibited the highest DH (91 ± 0.1%) and DPPH• radical scavenging activity (73.5 ± 0.25%) compared to the other hydrolysates. In addition, fractionation of the most effective (potent) hydrolysate by reverse phase high performance liquid chromatography indicated a direct association between hydrophobicity and radical scavenging activity of the hydrolysates. Isoelectric focusing tests also revealed that protein hydrolysates with basic and neutral isoelectric point (pI) have the highest radical scavenging activity, although few fractions in the acidic range also exhibited good antioxidant potential.     

The lipids of corn germ and endosperm

Mature kernels of an inbred corn were hand dissected into germ and endosperm fractions. Among various solvents tested, boiling, water-saturatedn-butanol extracted the most lipid from endosperm, and it was used as t h e extracting solvent for both germ and endosperm. The germ contained 78% of the total lipids and the endosperm 17%. The most striking differences in the fatty acid compositions of the triglycerides and polar lipids were higher levels of stearic and linolenic acids in the endosperm lipids. Although precautions were taken during extraction to inactivate lipases, immediately after harvest the free fatty acid level of the total lipids of the whole kernel was 6.5%. Ninety-five percent of the free fatty acids was in the endosperm fraction where the free fatty acids made up 36.5% of the total lipids. In germ, free fatty acids represented only 0.6% of the total lipids. The individual phospholipid and glycolipid classes of the endosperm and germ lipids were similar except for high levels of lyso compounds in the endosperm lipids. The higher levels of linolenic acid, free fatty acids and lyso lipids in endosperm may affect the keeping quality of the corn grain and of fractions milled from the endosperm. Presented at the AOCS meeting, St. Louis, May 1978.     

Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins

Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF). Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS) in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y) and tryptophan (W), in sequences identified by LC-MS as WYGPD (EYGF-23) and KLSDW (EYGF-33), contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56) was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69%) and IC₅₀ value (3.35 mg/mL). The SDNRNQGY peptide (10 mg/mL) had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL). In addition, YPSPV in (EYGF-33) (10 mg/mL) had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk.     

脱色猪血红蛋白酶解产物制备及其抗氧化性

通过复合蛋白酶(Pancreatin and Protamex)酶解制备了猪血红蛋白酶解产物,将其产物采用活性炭吸附脱色.酶解产物的分子量范围在＞15 kDa～＜1 kDa(游离氨基酸)之间,脱色酶解产物的分子量范围大部分＜5 kDa.脱色祛除94.36%的高铁血红素从而改善终产物的色泽.酶解产物及脱色产物具有还原力和DPPH自由基清除能力.猪血红蛋白酶解产物的抗氧化活性依赖于其分子量和氨基酸组成.