高效离子色谱法检测植酸钠的水解率

采用高效离子色谱（HPIC）法对植酸钠的水解产物进行分析,建立了HPIC法确定植酸钠水解率的新方法.用氢氧化钠、水和异丙醇组成的三元流动相梯度淋洗,植酸钠及其水解产物在不同的保留时间洗脱,经微膜抑制后进入电导检测器,15 min内可得检测结果,求出植酸钠的水解率.该方法用于确定植酸钠非酶水解时的水解率,收到了满意结果.     

An In-vitro Investigation of Selected Biological Activities of Hydrolysed Flaxseed (Linum usitatissimum L.) Proteins

A study was conducted to determine bioactivities of flaxseed (Linum usitatissimum L.; variety: Valour) proteins and their hydrolysates. Isolated flaxseed proteins were treated with Flavourzyme^® at different levels of enzyme to substrate ratio (E/S) and hydrolysis time. The unhydrolysed proteins and hydrolysates were studied for angiotensin I-converting enzyme inhibiting (ACEI) activity, hydroxyl radical (OH·) scavenging activity and bile acid binding ability. Flavourzyme catalysed hydrolysis generated hydrolysates with a 11.94–70.62% degree of hydrolysis (DH). The hydrolysates (0.67 mg/ml) had strong ACEI activity (71.59–88.29%). The maximum ACEI activity containing hydrolysate exhibited an IC₅₀ of 0.07 mg/ml (E/S: 1.5; Time: 12 h; DH: 11.94%). The OH· scavenging activity of the hydrolysates (0.5 mg/ml) was 12.48–22.08% with an IC₅₀ of 1.56 mg/ml in the sample possessing maximum activity (E/S: 47.5; Time 0.7 h; DH: 24.63%). Both these activities were greater in hydrolysates with lower DH and higher peptide chain length (PCL) than those with higher DH and lower PCL. Hydrolysed flaxseed proteins (0.67 mg/ml) had no bile acid binding ability. The unhydrolysed proteins had no ACEI or OH· scavenging activity but demonstrated bile acid binding ability.     

Investigation of hydrolysis conditions and properties on protein hydrolysates from flatfish skin

Response surface method (RSM), based on Box-Behnken design, was used to optimize the enzymatic hydrolysis conditions of flatfish skin protein hydrolysates (FSPH). Among the tested proteases, the combination of nutrase and trypsin was selected. The optimal hydrolysis conditions were as follows: pH 7.3, temperature 51.8°C, and the enzyme/substrate (E/S) ratio 2.5; under these conditions, the maximum peptide yield (PY) was 69.41±0.43%. The physiochemical analysis showed that the amino acids (His, Asp and Glu) of FSPH accounted for 18.15%, and FSPH was a mixture of polypeptides mostly distributed among 900–2000 Da. FSPH could exhibit a 93% chelating effect on ferrous ion at a concentration of 400 μg/mL, and also a notable reducing power. This study showed bioprocess for the production of FSPH for the first time, which had a good potential for valuable ingredients in the food, cosmetic and medicine industries.     

Peptide characteristics of sunflower protein hydrolysates

Protein isolate from sunflower seeds was used as the starting material for preparation of an extensively hydrolyzed peptide product. Protein was hydrolyzed using an endopeptidase (Alcalase), an exopeptidase (Flavourzyme), or both enzymes sequentially. Combined use of these proteases generated the highest degree of hydrolysis, 54.2%, and highest solubility, around 90%, between pH 2.5 and 7. Molecular weight profiles of the hydrolysates were characterized by gel filtration chromatography and denaturing electrophoresis. Amino acid compositions and solubilities of the different hydrolysates also were studied.     

Aqueous enzymatic extraction of peanut oil and protein hydrolysates

The aqueous enzymatic process of simultaneously preparing oil and protein hydrolysates from peanut was investigated. The optimum parameters for hydrolysis using Alcalase 2.4L were established by the single-factor and orthogonal test. The optimal processing conditions were as follows: hydrolysis temperature 60 °C, pH 9.5, ratio of material to water 1:5 (w/w), alkaline extraction time 90 min, enzyme amount 1.5% (w/w) and hydrolysis time 5 h. Under these conditions, the free oil and protein hydrolysates yields were 79.32% and 71.38% respectively. In order to improve these yields, As1398 was chosen to hydrolyze the residue and emulsion. The total free oil and protein hydrolysates yields were increased to 91.98% and 88.21% respectively.     

Enzyme-assisted extraction of chicken skin protein hydrolysates and fat: Degree of hydrolysis affects the physicochemical and functional properties

Enzyme-assisted extraction (EAE) of oils/fats involves the disruption of the cell wall of source material using enzymes to facilitate the release of oil. When proteases are used as the enzyme, EAE ends in the extracted oil as well as the protein hydrolysates. Herein, the EAE (using a commercial protease, Alcalase) was exploited to obtain fat and protein hydrolysates from chicken skin. Degree of hydrolysis (DH, the percentage ratio of cleaved peptide bonds), which showed a logarithmic correlation with the reaction time, was found to affect the properties of the products. As the DH increased, the peptide chain length of protein hydrolysates decreased which was confirmed by SDS-PAGE analysis. With the increase of DH, the emulsifying activity index, foaming capacity, and oil holding capacity of the hydrolysates decreased but the solubility and emulsion stability index increased (p < 0.05). The DPPH free radical scavenging activity of the hydrolysates increased with the DH up to DH = 39.62% but decreased thereafter (p < 0.05). EAE resulted in a rise in fat yield and the fat contained a higher amount of unsaponifiables and lower free fatty acids (FFA) content, as compared to the control treatment (No enzyme, 80°C, 2 h, p < 0.05). DH affected the fat yield and the unsaponifiables content of the fat, positively (p < 0.05). However, it did not affect the fat FFA content and iodine value (p > 0.05). Results obtained here showed DH can be used as an effective measure for controlling the physicochemical and functional properties of chicken skin protein hydrolysates and fat in the EAE process.     

Hydrolysis of aromatic heterocyclic polymers in high temperature water. I. Hydrolysis of polyphenyl-1,2,4-triazine

The hydrolytic behaviors of polyphenyl-1,2,4-triazine (As–PPT) at high temperatures were investigated both experimentally and theoretically. The hydrolytic experiments of As–PPT film and a reasonably designed model compound for the polymer were carried out in distilled water at 250°C/3.97 MPa under N₂ atmosphere, respectively. The hydrolytic reactions were monitored by FTIR and UV-Vis spectra, and their hydrolysates were identified by FTIR, HPLC, and MS. The results confirmed that the two reactions gave the same major hydrolysate, terephthalic acid. In addition, the electronic structure of its model compound, given by ab initio calculation at the 4-31G level, indicated that the triazine ring is the hydrolytically active part at which the N atoms are to be protonated. On the basis of these results, a hydrolytic mechanism is proposed, which suggests that the cleavage of one C N bond takes place, primarily on the triazine rings, followed by a gradual rupture of the polymer chain and the formation of the intermediate terephthalonitrile, the further hydrolysis of which produces terephthalic acid.© 2001 John Wiley & Sons, Inc. J Appl Polym Sci 82;907–915;2001     

Enzymatic modification of proteins of commercial sesame meals (Sesamum indicum, L.)

Sesame (Sesamum indicum, L.) is one of the most important oilseed crops in Venezuela. However, the low solubility of the flour made of commercial meals does not allow its use in the preparation of fluid foods. To solve this situation, the alternative of solubilizing the sesame proteins by an enzymatic method, using commercial proteases, was studied. Hydrolysis was carried out with two types of bacterial proteases: neutrasa 0.5L, and alcalase 0.6L. Basically, the process consisted of milling and sieving the sesame cake (60 mesh), concentrating the proteins by solubilizing them at a pH of 9.5, and then precipitating at a pH of 4.5. Proteins were hydrolyzed by an enzymatic method, and the hydrolysates freeze-dried and spray-dried. Optimal conditions of hydrolysis using neutrase 0.5L at a pH of 7 were: 6% substrate concentration, 3% enzyme:substrate ratio, temperature 50 degrees +/- 1 degree C, and hydrolysis degree of 8%. When alcalase 0.6L was used at a pH of 8, optimal conditions were: 8% substrate concentration, 2.3% enzyme:substrate ratio, temperature 58 degrees +/- 1 degree C, and hydrolysis degree of 10%. The enzyme affinity (Km) was best at temperatures near the optimal temperature for the hydrolysates. Dried hydrolysates had protein values which ranged between 66.3% and 66.9% and a nitrogen solubility in water, of approximately 85%. Hydrolysis yields referred to the concentrate dried mass were 42% for the atomized hydrolysate, and 56% for the freeze-dried one. In conclusion, the enzymatic hydrolysis process improved the nitrogen solubility in water of the sesame proteins.     

Enhancing ACE-inhibition of food protein hydrolysates by selective adsorption using porous carbon materials

Bioactive peptides from food proteins such as natural ACE (angiotensin-converting enzyme)-inhibitors have attracted particular attention for their potential to prevent hypertension. ACE-inhibiting peptides were enriched from food protein hydrolysates prepared from α-lactalbumin and lysozyme by selective adsorption on microporous activated carbons. For the eluate, it was shown by liquid chromatography that the strongest inhibitor isoleucyl-tryptophan was enriched by a factor of 11.2 compared to the initial α-lactalbumin hydrolysate. Natural inhibitors derived from lysozyme hydrolysates (e.g., alanyl-tryptophan) were successfully enriched as well. Identification of the enriched peptide fraction by mass spectroscopy revealed the hydrophobic character of the enriched peptides. The molecular weight distribution of the enriched peptide fraction can be controlled by the pore size distribution of the chosen adsorbent, which was proven by size exclusion chromatography of enriched peptide fractions derived from three different model carbons differing in their pore size. The selective enrichment of natural ACE-inhibitors from the α-lactalbumin hydrolysate lead to a 6 times stronger in vitro ACE-inhibition demonstrating the high potential as ingredients for hypotensive functional foods with reduced side effects.     

In Vitro Acetylcholinesterase-Inhibitory Properties of Enzymatic Hemp Seed Protein Hydrolysates

The aim of this work was to characterize the structural and functional properties of hemp seed protein‐derived acetylcholinesterase (AChE)‐inhibitory enzymatic hydrolysates. Hemp seed protein isolate hydrolysis was performed using six different proteases (pepsin, papain, thermoase, flavourzyme, alcalase and pepsin + pancreatin) at different concentrations (1–4 %). The degree of hydrolysis was directly related to the amount of protease used but had no relationship with AChE‐inhibitory activity. Amino acid composition results showed that the hemp seed protein hydrolysates (HPHs) had high levels of negatively charged amino acids (39.62–40.18 %) as well as arginine. The 1 % pepsin HPH was the most active AChE inhibitor with ~6 µg/mL IC₅₀ value when compared to 8–11.6 µg/mL for the other HPHs. Mass spectrometry analysis showed that most of the peptides in all the hydrolysates were less than 1000 Da in size. However, the pepsin HPHs contained larger‐sized peptides (244–1009 Da) than the papain HPHs (246–758 Da), which in turn was larger than the alcalase HPH (246–607 Da). The higher AChE‐inhibitory effects of the pepsin HPHs may be due to increased synergistic effects from a wider peptide size range when compared to the papain and alcalase HPHs that had narrower ranges. The narrow peptide size range in the alcalase HPH confirms the higher efficiency of this protease in releasing small‐sized peptides from food proteins.     

Characterization of Peptides Found in Unprocessed and Extruded Amaranth (Amaranthus hypochondriacus) Pepsin/Pancreatin Hydrolysates

The objectives of this study were to characterize peptides found in unprocessed amaranth hydrolysates (UAH) and extruded amaranth hydrolysates (EAH) and to determine the effect of the hydrolysis time on the profile of peptides produced. Amaranth grain was extruded in a single screw extruder at 125 °C of extrusion temperature and 130 rpm of screw speed. Unprocessed and extruded amaranth flour were hydrolyzed with pepsin/pancreatin enzymes following a kinetic at 10, 25, 60, 90, 120 and 180 min for each enzyme. After 180 min of pepsin hydrolysis, aliquots were taken at each time during pancreatin hydrolysis to characterize the hydrolysates by MALDI-TOF/MS-MS. Molecular masses (MM) (527, 567, 802, 984, 1295, 1545, 2034 and 2064 Da) of peptides appeared consistently during hydrolysis, showing high intensity at 10 min (2064 Da), 120 min (802 Da) and 180 min (567 Da) in UAH. EAH showed high intensity at 10 min (2034 Da) and 120 min (984, 1295 and 1545 Da). Extrusion produced more peptides with MM lower than 1000 Da immediately after 10 min of hydrolysis. Hydrolysis time impacted on the peptide profile, as longer the time lower the MM in both amaranth hydrolysates. Sequences obtained were analyzed for their biological activity at BIOPEP, showing important inhibitory activities related to chronic diseases. These peptides could be used as a food ingredient/supplement in a healthy diet to prevent the risk to develop chronic diseases.     

Analyzing molecular weight distribution of whey protein hydrolysates

Process parameters on enzymatic hydrolysis and molecular weight (MW) distribution of whey protein hydrolysates were investigated. Whey protein hydrolysates were first gained by the alkaline protease alcalase for 7 h at temperature (50 °C), pH (8.0) and E/S (3%). The diversification of the hydrolysis degree and dissociative amino acid content was investigated during the whey hydrolysis. The dissociative amino acid content was 56.09 μmol/mL with the hydrolysis degree of 20.04%. The results of Sephadex G25 washing and high performance liquid chromatography–electrospray ionization–mass spectrometry (HPLC–ESI–MS) indicated the molecular weight distribution of whey protein hydrolysates ranged from 300 to 1400 Da, and most of whey peptide was under 1000 Da.     

Isolation of an active peptide fragment from human serum albumin and its synergism with α-tocopherol

An active peptide was isolated from hydrolysates of human serum albumin. This peptide was initially isolated by gel permeation chromatography and subsequent reversed-phase high-performance liquid chromatography. This active peptide, composed of 10 amino acid residues, was further hydrolyzed with a lysyl endopeptidase to give two peptide fragments. Only one fragment, identified as the tetrapeptide Leu-Gln-His-Lys, was found to have activity comparable to the original peptide and corresponded to the amino acid residues 103–106 of human serum albumin. Among these four amino acid residues, the His-Lys sequence seemed to be important in the occurrence of potent activity by comparison of the structural similarity with another active tetrapeptide, Asp-Thr-His-Lys, which had been previously isolated from bovine serum albumin hydrolysates. In addition, the active fragment showed potent synergism by preventing consumption of α-tocopherol during the autoxidation of linoleic acid.     

Isolation of an active peptide fragment from human serum albumin and its synergism with α-tocopherol

An active peptide was isolated from hydrolysates of human serum albumin. This peptide was initially isolated by gel permeation chromatography and subsequent reversed-phase high-performance liquid chromatography. This active peptide, composed of 10 amino acid residues, was further hydrolyzed with a lysyl endopeptidase to give two peptide fragments. Only one fragment, identified as the tetrapeptide Leu-Gln-His-Lys, was found to have activity comparable to the original peptide and corresponded to the amino acid residues 103–106 of human serum albumin. Among these four amino acid residues, the His-Lys sequence seemed to be important in the occurrence of potent activity by comparison of the structural similarity with another active tetrapeptide, Asp-Thr-His-Lys, which had been previously isolated from bovine serum albumin hydrolysates. In addition, the active fragment showed potent synergism by preventing consumption of α-tocopherol during the autoxidation of linoleic acid.     

Optimization of the Aqueous Enzymatic Extraction of Rapeseed Oil and Protein Hydrolysates

An aqueous enzymatic extraction method was developed to obtain free oil and protein hydrolysates from dehulled rapeseeds. The rapeseed slurry was treated by the chosen combination of pectinase, cellulase, and β-glucanase (4:1:1, v/v/v) at concentration of 2.5% (v/w) for 4 h. This was followed by sequential treatments consisting of alkaline extraction and an alkaline protease (Alcalase 2.4L) hydrolysis to both produce a protein hydrolysate product and demulsify the oil. Response surface methodology (RSM) was used to study and optimize the effects of the pH of the alkaline extraction (9.0, 10.0 and 11.0), the concentration of the Alcalase 2.4L (0.5, 1.0 and 1.5%, v/w), and the duration of the hydrolysis (60, 120, and 180 min). Increasing the concentration of Alcalase 2.4L and the duration of the hydrolysis time significantly increased the yields of free oil and protein hydrolysates and the degree of protein hydrolysis (DH), while the alkaline extraction pH had a significant effect only on the yield of the protein hydrolysates. Following an alkaline extraction at pH 10 for 30 min, we defined a practical optimum protocol consisting of a concentration of 1.25–1.5% Alcalase 2.4L and a hydrolysis time between 150 and 180 min. Under these conditions, the yields of free oil and protein hydrolysates were 73–76% and 80–83%, respectively. The hydrolysates consisted of approximately 96% of peptides with a MW less than 1500, of which about 81% had a MW less than 600 Da.     

猪皮水解严物的吸湿保湿性能研究

研究猪皮水解产物的吸湿保湿性能。实验利用动物蛋白酶对猪皮进行酶解,以骨胶原为对照,对猪皮水解产物在26℃,相对湿度分别为20％、43％和81％的条件下的吸湿和保湿性能进行测定。在实验湿度环境范围内,猪皮在水煮45min后进行酶解,酶解条件为底物质量分数为4％、酶浓度为2％、酶解温度40％、酶解时间2h,pH值7．5的酶解条件下,酶解产物得率最高,酶解产物的吸湿和保湿性能和德国产的骨胶原相近。猪皮的动物蛋白酶水解产物用于开发吸湿保湿化妆品方面具有极大的开发前景。     

Hydrolysis of pumpkin oil cake protein isolate and free radical scavenging activity of hydrolysates: Influence of temperature,enzyme/substrate ratio and time

The effect of hydrolysis parameters (temperature, initial enzyme/substrate ratio and time) on the hydrolysis of pumpkin oil cake protein isolate (PuOC PI) with acid protease from Aspergillus niger and the antioxidant potency of the obtained hydrolysates were studied by response surface methodology (RSM). The hydrolysis progress, measured by the degree of hydrolysis (DH), was described by a second-order polynomial model (R² = 0.77) and the conditions for optimum DH (42.94%) were found at temperature of 40 °C, enzyme/substrate ratio (E/S) 4.38 HUT/mg of substrate proteins and 85 min. The antiradical activity (AA) of the PuOC PI hydrolysates was examined by the 1,1-diphenyl-2-picryl-hydrazyl (DPPH) assay; all hydrolysates showed a concentration dependent scavenging activity against DPPH radicals. The AA of hydrolysates was influenced by process parameters and was presented also by a second-order polynomial model (R² = 0.7). The conditions to achieve the highest DH did not result hydrolysates with the optimum AA; the highest AA ranged from 34% to 40% and were found in hydrolysates obtained at 50 °C.     

Enzymatic modification of sesame seed protein,sourced from waste resource for nutraceutical application

The functional characteristics which include protein solubility at different pH, emulsifying and foaming properties, degree of hydrolysis, molecular weight distribution, antioxidant and ACE inhibitory activity of sesame protein hydrolysates prepared with pepsin, papain and alcalase enzymes were evaluated. The rate of degree of hydrolysis was found to reach maximum (25–30%) within the first time fragment i.e 10 min but 80% of hydrolysis was obtained in 120 min with alcalase. SDS-PAGE of hydrolysates with papain, pepsin and alcalase evinced bands of low molecular weight protein of 14.3 kDa and even lower for alcalase treatment of 120 min. Hydrolysates so formed were of improved functional properties as evident from emulsifying and foaming property. Hydrolysis with different proteases enhanced the protein solubility significantly at pH 7.0. Antioxidative assay revealed radical scavenging activity of the hydrolysates with papain hydrolysates showing maximum antioxidative efficacy. The ultra-filtered peptide fractions which showed comparable ACE inhibitory activity were sequenced by MALDI-TOF and matched to that of previously identified ACE inhibitory peptides. The results corroborate the ACE inhibitory effect of the peptides. Hence, these highly bioactive protein hydrolysates produced from waste sesame meals can be successfully employed in various functional food formulations.     

Sugarcane bagasse hydrolysis with phosphoric and sulfuric acids and hydrolysate detoxification for xylitol production

The effectiveness of phosphoric acid to release xylose from sugarcane bagasse hemicellulose was assessed through a 2³ full factorial design. The maximum xylose concentration in the hydrolysate (17.1 g dm^?3) was attained when the bagasse was treated at 160 °C for 60 min, using 70 mg of phosphoric acid per gram of dry‐bagasse. Hydrolysis carried out with sulfuric acid, under optimum conditions previously determined, provided a hydrolysate with a similar xylose concentration (17.2 g dm^?3). After vacuum concentration, these hydrolysates were detoxified and used for xylitol production with the yeast Candida guilliermondii. Two different detoxification strategies, which consisted of adjusting the pH of the hydrolysates to 5.5 with either calcium oxide or ammonium hydroxide, both followed by active charcoal adsorption, were tested. The best xylitol productions (18.1 and 19.2 g dm^?3) were observed when calcium oxide was used to adjust the pH of both the phosphoric and the sulfuric acid hydrolysates, respectively. Copyright © 2004 Society of Chemical Industry     

Hydrolytic degradation of cured urea–formaldehyde resin

The degradation of cured urea–formaldehyde (UF) resin in aqueous suspension was investigated by gravimetric analysis of the changes in the content of nonextractable low-molecular components. In acid conditions (pH 4.0) at 47°C the consecutive processes of post-curing and of polymer break-down (activation energy 90 kJ/mol) are detectable whereas at 80°C and 97°C only the formation of the extractable hydrolysates is observed. The degraded polymer contains less carbonyl groups than does the original resin substrate as shown by means of infrared (IR) analysis. In contrast to the results of the tests carried out in acid conditions, in neutral and basic aqueous media the hydrolytic decomposition of UF macromolecular network is less significant. During the hydrolysis of UF polymer at 30°C–45°C the concentration of formaldehyde released from the resin to the aqueous phase increases initially (2 days) at a relatively high rate both at acid and alkaline pH. Then its growth slows down but is still detectable in acid conditions, whereas in basic medium no further liberation of HCHO is observed.