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1.
To observe the role of sarcoplasmic protein (SP) on myofibrillar protein (MP) denaturation under a hydrostatic pressure (HP), MP isolated from bovine muscle was treated with 300 MPa by increasing concentrations of SP (0, 0.8, 1.6, and 3.2 mg/ml) from bovine. SDS-PAGE patterns of soluble proteins in 0.1M NaCl (pH 7.4) indicated that a protein (about 100 kDa) from MP decreased with increasing concentrations of SP and that a 97 kDa protein from SP observed with 0.1 MPa was not observed with 300 MPa. SDS-PAGE patterns of soluble proteins in 0.6 M NaCl (pH 7.4) and Ca-ATPase activity showed that the denaturation of myosin heavy chain (MHC) was accelerated with increasing SP concentrations with the 300 MPa treatment. Thus, the addition of SP enhanced HP-induced denaturation of MHC and of a protein from MP of about 100 kDa. 相似文献
2.
Effect of high pressure treatment on aggregation and structural properties of soy protein isolate 总被引:3,自引:0,他引:3
The aggregation and structural properties of soy protein isolate (SPI), induced by high pressure (HP) treatment at 200-600 MPa were investigated by size-exclusion chromatography combined with multi-angle laser scattering (SEC-MALLS) and fourier transform infrared (FTIR) spectroscopy. HP treatment at lower pressure level (e.g., 200 MPa) resulted in formation of marked insoluble aggregate of SPI, while the treatment at higher pressure level (e.g., 600 MPa) led to transformation of insoluble aggregate to soluble one. The soluble aggregate formed at 400 or 600 MPa had much less mean molecular weight (MW) (about 5.2 × 106 g/mol) than that at 200 MPa (about 1.6 × 107 g/mol), and was also much more homogenous in MW distribution. FTIR analyses confirmed changes in secondary and tertiary structures, induced by HP treatment. These results can provide direct evidence or explanation for HP-induced modification of soy proteins. 相似文献
3.
Rong He Hai-Yan He Dongfang Chao Xingrong Ju Rotimi Aluko 《Food and Bioprocess Technology》2014,7(5):1344-1353
High pressure (HP, 200, 400, and 600 MPa)- and heat (60, 80, and 100 °C)-induced gelation, aggregation, and structural conformations of rapeseed protein isolate (RPI) were characterized using gel permeation–size-exclusion chromatography, differential scanning calorimetry, and circular dichroism (CD) techniques. HP treatments significantly (p?<?0.05) increased the content of soluble protein aggregates and surface hydrophobicity of RPI. In contrast, heat treatments at 80 and 100 °C led to significant (p?<?0.05) decreases in the amount of soluble protein aggregates. At pressure treatment of 200 MPa, there was a significant (p?<?0.05) increase in free sulfhydryl group content of RPI, whereas 400- and 600-MPa treatments as well as temperature treatments (60–100 °C) caused significant decreases. Protein denaturation temperature was increased by about 6 °C by HP and heat treatments. The far-UV CD spectra revealed increases in α-helix content of RPI after HP treatments with 400 MPa producing the most increase. Near-UV data showed that HP and heat treatments of RPI led to increasing interactions among the aromatic amino acids (evidence of protein aggregation), and between aromatic amino acids and the hydrophilic environment, which indicates protein unfolding. Least gelation concentration of RPI was significantly (p?<?0.05) reduced by HP and heat treatments, but HP-treated RPI produced gels with better textural properties (hardness increased from ~7.7 to 81.1 N, while springiness increased from ~0.37 to 0.99). Overall, pressure treatments (200–600 MPa) were better than heat treatments (60–100 °C) to modify the structure and improve gelation properties of RPI. 相似文献
4.
Mingchun Lv Beibei Tan Rong Yang Anqi Xu Jinjie Zhang Dalun Xu Wenge Yang 《International Journal of Food Science & Technology》2020,55(4):1515-1522
The effects of high-pressure (HP) treatment parameters (300 MPa for 5 min, 350 MPa for 3 or 5 min and 400 MPa for 1 min) on the biochemical properties and structure of myofibrillar protein (MP) from Tegillarca granosa were investigated. The results showed that HP significantly affected the biochemical properties of MP. Under the HP treatment at 400 MPa for 1 min, the MP extraction yield was reduced from 32.21 to 18.34 mg g−1, and its surface hydrophobicity was increased from 89.29 to 104.69 µg. Ca2+-ATPase activity of MP decreased from 0.019 to 0.011 µmol (pi) mg−1 (pro) min−1, and both total and reactive sulfhydryl contents significantly declined. Furthermore, HP caused MP to undergo conformational changes. This study provides a reference data for the processing of T. granosa using HP treatment. 相似文献
5.
Mahesh Venkatachalam Shridhar K Sathe 《Journal of the science of food and agriculture》2007,87(8):1539-1549
Val bean (Lablab purpureus L.) proteins were fractionated using the Osborne protein fractionation scheme and biochemically characterized. The seed flour contained 302 g kg?1 protein (micro‐Kjeldahl N × 6.25) on a dry weight basis. Albumin, globulin, prolamin, and glutelin accounted for 22.8%, 45.1%, 1.8% and 30.3%, respectively, of the total soluble seed proteins. Among the solvents tested, 0.1 mol L?1 aqueous NaOH was the most effective protein solubilizer. Isoelectric focusing indicated the seed proteins to be predominantly acidic (pI range was ~4–7). Val globulin is a glycoprotein composed of at least three polypeptides in the molecular mass range 51–64 kDa. Albumin fraction had the highest trypsin inhibitory activity, while the globulin fraction registered the highest hemagglutinating activity. Sulfur amino acids were the first limiting amino acids in the total seed proteins. The proportion of essential to total [E/T(%)] amino acids for the bean flour was 36.97%. Among the protein fractions, glutelin fraction had the highest E/T (42.86%) followed by albumin (41.57%), globulin (39.87%), and prolamin (39.15%). Native globulin, although resistant to pepsin, was effectively digested in vitro upon moist heat (100 °C, 30 min) denaturation. Copyright © 2007 Society of Chemical Industry 相似文献
6.
Nicolas Chapleau Ccile Mangavel Jean‐Pierre Compoint Marie de Lamballerie‐Anton 《Journal of the science of food and agriculture》2004,84(1):66-74
Modification of myofibrillar proteins induced by high‐pressure processing has been investigated at pressures ranging from 50 to 600 MPa for 10 min at 20 °C. Analysis by spectroscopic methods and circular dichroism of myofibrillar proteins in phosphate buffer pH 6.0 containing 0.6 M KCl showed no changes in the secondary structure of proteins. However, study of protein conformation by quasielastic light scattering and gel filtration chromatography proved the emergence of aggregation after treatment at pressures higher than 300 MPa. This aggregation was accompanied by enhanced binding of anilino‐1‐naphthalene‐8‐sulphonic acid, which indicated an increase in hydrophobic bonding of myofibrillar proteins. Modification of the tertiary and quaternary structures of proteins may induce a molten globule state. Copyright © 2003 Society of Chemical Industry 相似文献
7.
Shashi Kiran Jasra Barbara Zielinski Pardeep Kumar Jasra 《Journal of the science of food and agriculture》2006,86(15):2609-2620
The effects of various cryoprotective additives separately and in combination were studied on the myofibrillar protein integrity, biochemical enzyme activity levels and muscle ultrastructure in the freshwater teleost fish Rohu carp (Labeo rohita). Fish muscle samples were divided into eight groups and immersed in different mixtures of cryoprotective additives (S1–S8), then frozen at ? 20 or ? 30 °C for 24 months. Electrophoretic studies revealed early (within 6 months) alteration of the myofibrillar proteins myosin light chain, α‐actinin and tropomyosin. Reduction of the storage temperature from ? 20 to ? 30 °C slowed down the degradative processes. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that fish muscle treated with cryoprotective mixture S8 (40 g L?1 sorbitol/3 g L?1 sodium tripolyphosphate/4 g L?1 sodium alginate) showed minimal post mortem changes in myofibrillar proteins. Ultrastructural results also revealed post mortem damage to the muscle, seen earliest (within 6 months) in the sample frozen‐stored without additives (S2), as compared with the normal, unfrozen muscle (S1). The influence of cryoprotectants alone and in combination on fish muscle structural proteins, myosin and actin filaments (A and I bands), during prolonged frozen storage was investigated. After 12 months, samples frozen‐stored with various cryoprotective additives (S2‐S7), except S8, showed signs of myofibrillar disintegration. Beyond that time the degradative processes started showing up in all samples, with minimal muscle ultrastructural damage in sample S8. Again, reducing the storage temperature from ? 20 to ? 30 °C slowed down the degradative processes. Ultrastructural results correlated well with levels of biochemical enzymes (Ca2+ myofibrillar ATPase and succinic dehydrogenase) during frozen storage. This is the first report of the cryoprotective effects of these additives on this popular edible fish species. Of the various combinations of additives tested, cryoprotective mixture S8 was found to preserve the muscle structure longest under frozen storage conditions. However, even this mixture was only effective for 18 months at ? 30 °C. Beyond that time the myofibrillar degradative processes were apparent with correlative electrophoretic, biochemical and ultrastructural studies. Copyright © 2006 Society of Chemical Industry 相似文献
8.
Johannes de Bruijn José Martínez‐Oyanedel Cristina Loyola Javier Seiter Francisco Lobos Ricardo Pérez‐Arias 《International Journal of Food Science & Technology》2011,46(8):1691-1698
After ultrafiltration (UF) thermally unstable compounds of Sauvignon blanc wines were concentrated in the retentate fractions of 10–30 kDa increasing heat‐induced haze 8.9‐fold, while concentrating haze stabilising species into the retentate fractions of 100–300 kDa with a 5.3‐fold reduction of heat‐induced haze compared to unfiltered wine. Proteins having molecular weights of 18, 23, 33, 35, 40 and 60 kDa were identified as the main haze promoting agents for Sauvignon blanc, where the proteins of 56, 69 and 72 kDa were identified as thermally stable. Potassium sulphate (100 mg L?1) and magnesium sulphate (25 mg L?1) did not affect wine haziness at experimental diafiltration (DF) conditions using model wine solutions. Combination of UF and DF is a useful separation method of wine proteins, which is helpful to quantify haziness of individual species and to reveal the interactions between species involving protein haze. 相似文献
9.
Understanding phase transition during high‐pressure (HP) processing of foods is important both with respect to optimizing the process and improvement of product quality, but scientific information available in this area is very limited. In this study, the phase‐transition behavior of water was evaluated using a HP differential scanning calorimetry (DSC). Tests were carried out under both isothermal pressure‐scan (P‐scan) and isobaric temperature‐scan (T‐scan) modes with distilled water prefrozen in the sample cell. P‐scan was carried out at 0.3 MPa/min at two temperatures, ?10 and ?20C, and T‐scan was carried out at 0.15C/min at two pressures, 0.1 and 115 MPa. The pressure‐induced phase transition of water was accurately reproduced by the P‐scan test. Ice melting latent heat during P‐scan showed no significant difference (P > 0.05) from the available reference data in literature. The relationship between P‐scan tested (Lm) and reference latent heat was Lm = 0.987 L (R2 = 0.99, n = 6) suggesting a mean error less than 2%. T‐scan mode was less appropriate and did not yield promising result. Measured values were less accurate than P‐scan probably because of the influence of large heat capacity of sample cell. However, reliable and reproducible results obtained under P‐scan mode suggested that the HP DSC can be used for the calorimetric determination of pressure‐dependent water‐phase transition in real food systems during HP freezing/thawing operations. 相似文献
10.
M.C. Añón M. de Lamballerie F. Speroni 《Innovative Food Science and Emerging Technologies》2011,12(4):443-450
In the present work the effect of high pressure (HP) treatment in the presence of NaCl on the thermal behavior of soybean proteins was analyzed by differential scanning calorimetry. The thermograms obtained have shown that NaCl addition increased the thermal stability – increase in temperatures of denaturation (Td) – of both glycinin and β-conglycinin. HP treatments increased thermal stability of glycinin, but decreased that of β-conglycinin. High NaCl concentrations decreased (in glycinin) or inverted (in β-conglycinin) the effects of HP on thermal stability. Cooperativity of denaturation of glycinin was enhanced by NaCl and HP. Cooperativity of denaturation of β-conglycinin was enhanced by HP and also by NaCl at 0.2 mol/L but decreased with the combination of treatments. Salt addition increased the enthalpy, ΔH, of denaturation of glycinin and β-conglycinin, being this effect stronger on glycinin. HP treatment provoked the denaturation of both protein fractions. The presence of NaCl protected glycinin against HP-denaturation at any assayed salt concentration and pressure level, while β-conglycinin was only protected at 200 and 400 MPa, but was more denaturated at 600 MPa in the presence of 0.6 mol/L of NaCl.
Industrial relevance
The knowledge provided by this work may be useful in the handling of high pressure-treated food with high NaCl content (e.g. meat emulsions, smallgoods) where soybean proteins are used as additives, in order to choose high pressure values to achieve their denaturation or predict the effects of ulterior thermal treatments. Thus, this knowledge may be useful to increase the use of high pressure in food industry. 相似文献11.
Thazhakot V Sankar Alappat Ramachandran 《Journal of the science of food and agriculture》2005,85(4):563-568
The characteristics and stability of natural actomyosin (NAM) from rohu (Labeo rohita), catla (Catla catla) and mrigal (Cirrhinus mrigala) were investigated. The total extractable actomyosin (AM) was higher (7.60 mg ml?1) in the case of rohu compared with that from catla and mrigal (5 mg ml?1). Although the specific AM ATPase activity was similar (0.43–0.5 µmol P min?1 mg P?1) among the three species, the total ATPase activity was lower in mrigal (25 µmol g?1 meat) compared with the other species (37 µmol g?1 meat). The inactivation rate constants (kd) of AM Ca ATPase activity showed differences in the stabilities of actomyosin among these fish, the actomyosin from catla being least stable. The NAM from these species was stable up to 20 °C at pH 7.0. Catla AM became unstable at 30 °C, while rohu and mrigal AM could withstand up to 45 °C. The thermal denaturation with respect to solubility, turbidity, ATPase activity, sulphhydryl group and surface hydrophobicity showed noticeable changes at around these temperatures. Copyright © 2004 Society of Chemical Industry 相似文献
12.
Synergistic effects of high pressure processing and slightly acidic electrolysed water on the inactivation of Bacillus cereus spores
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Liping Wang Qiang Xia Yunfei Li 《International Journal of Food Science & Technology》2017,52(11):2429-2435
Bacillus spores are concerns for their resistance to heat, high pressure processing (HPP), and disinfectants. We examined the effects of HPP and slightly acidic electrolysed water (SAEW) on inactivation of B. cereus spores. Spores' suspensions were prepared with 2‐(N‐morpholino) ethanesulfonic acid (MES) buffer or SAEW with available chlorine content (ACC) of 24, 35, 44 or 55 mg L?1, and then subjected to HPP. The individual effects of HPP or SAEW on spores were negligible (<1.0 log CFU mL?1). With factorial design and anova analysis, HPP + SAEW treatment was shown to have significantly positive effects on spores’ inactivation. The optimal conditions were 300 MPa HPP + SAEW with 44 mg L?1 ACC or 200 MPa HPP + SAEW with 44 mg L?1 ACC + 500 MPa HPP, producing reductions of 3.27 and 3.99 log CFU mL?1, respectively. HPP + SAEW have potentials to serve as two effective hurdle techniques for inactivating B. cereus spores. 相似文献
13.
Southern Ocean swimming crab Ovalipes trimaculatus and the Patagonian stone crab Platyxanthus patagonicus are fishing resources with commercial value. Thermal treatment of crabs is necessary to denature muscle proteins, facilitating meat detachment from the crab shell (picking procedure). The proximal composition, protein patterns of crab muscle, thermophysical properties and heat transfer coefficients were determined. Heat transfer during thermal processing of body (i.e., cephalothorax) and claws of both crab species was simulated using a finite element computational code; the simulations were experimentally validated. Color changes in crab muscle during the heating process were measured. Thermal denaturation kinetics of myofibrillar proteins was determined using Differential Scanning Calorimetry (DSC) in small samples previously heated in water under controlled conditions. DSC thermograms of raw crab muscle showed two peaks at 49.0 ± 0.4 and 77.5 ± 0.6 °C corresponding to myosin and actin respectively. Activation energies for the denaturation of myosin (145.70 kJ/mol) and actin (156.42 kJ/mol) were calculated from Arrhenius equation. The degree of denaturation achieved by the myofibrillar proteins at the coldest point of the muscle in body and claws during the heating process was established by considering the protein denaturation kinetics determined by DSC, the activation energies and the heat penetration curves. Adequate conditions for the detachment of meat from the crab exoskeleton were established. The obtained results may help in determining the optimal heating times during the industrialization of these crustaceans. 相似文献
14.
Erdal S 《Journal of the science of food and agriculture》2012,92(4):839-843
BACKROUND: Mammalian sex hormones (MSH)—progesterone, β‐estradiol and androsterone—enhance plant growth and development by stimulating significant morphological and biochemical parameters under normal conditions. However, there is no report regarding their effects on plants exposed to environmental stress conditions. Therefore, the present study was focused on elucidating the possible positive effects of MSH on seedling growth, antioxidant activity and synthesis reactions in maize seeds exposed to salt stress, one of the most important environmental stresses. For this purpose, the various concentrations (10?6, 10?8, 10?10 and 10?12 mol L?1) of MSH were studied. RESULTS: Salinity (100 mmol L?1 NaCl) significantly reduced root length and seedling height, whereas MSH treatment significantly ameliorated the adverse effects of salinity on root length and seedling height. On the other hand, although salinity increased soluble protein, soluble sugar and proline content in 7‐day‐old maize seedlings, these were higher in MSH‐treated seedlings. Similarly, MSH treatment augmented superoxide dismutase, peroxidase and catalase activities under salt stress, whereas it decreased superoxide production and lipid peroxidation level. The most favorable concentrations were determined as 10?8 mol L?1 for progesterone and β‐estradiol and 10?10 mol L?1 for androsterone. CONCLUSION: Exogenous MSH application was found to have an important ameliorative effect on growth of seeds exposed to salt stress by stimulating antioxidant activity and synthesis reactions. This is the first study investigating the effects of MSH on germination of seeds exposed to stress conditions. Copyright © 2011 Society of Chemical Industry 相似文献
15.
Theocharis Chatzistathis Ioannis Therios Gregorios Diamantidis 《Journal of the science of food and agriculture》2010,90(4):676-682
BACKGROUND: The time course of polyphenol oxidase (PPO) activity in the leaves of two olive cultivars (Picual and FS‐17) irrigated with nutrient solutions differing in Mn concentration (0, 2 and 1280 µmol L?1) was studied under hydroponic conditions to determine whether PPO activity could be used as an early criterion of Mn status of olive plants, and to elucidate whether genotypic differences exist between the two olive cultivars studied, concerning the effect of Mn concentration on PPO activity. RESULTS: In all the Mn treatments, PPO activity was greater in Picual than in FS‐17. Under excess Mn (1280 µmol L?1), PPO activity gradually increased with time, starting from day 30 of the experiment in both cultivars, and this increase preceded the appearance of Mn toxicity symptoms. In contrast, in the other two Mn treatments (0 and 2 µmol L?1) PPO activity increased and afterwards decreased during the experiment, but the trend was not clear. In the 1280 µmol L?1 treatment, PPO activity linearly increased (R = 0.8836 for Picual and 0.943 for FS‐17) with the increase of Mn concentration in the leaves of both cultivars. In the 1280 µmol L?1 Mn treatment, PPO activity was negatively related with Fe and Zn concentrations in the leaves, and positively in the 0 and 2 µmol L?1 Mn treatments with the Ca, Mg and K concentrations. CONCLUSION: From the differential time course of PPO activity in the three Mn treatments (0, 2 and 1280 µmol L?1), it is concluded that periodic measurements of PPO activity in the leaves of the olive cultivars Picual and FS‐17 can be used for the early detection of Mn toxicity (before the appearance of symptoms). Copyright © 2010 Society of Chemical Industry 相似文献
16.
Dimuthu Bogahawaththa Biljana Trajkovska Tatijana Markoska Todor Vasiljevic 《Journal of dairy science》2021,104(3):2834-2842
Heating, pressurization, and shearing can modify native milk proteins. The effects of pressurized heating (0.5 vs. 10 MPa at 75 or 95°C) with shearing (1,000 s?1) on proteins of raw bovine skim milk (SM, ~9% total solids) and concentrated raw skim milk (CSM, ~22% total solids) was investigated. The effects of evaporative concentration at 55°C and pressurized shearing (10 MPa, 1,000 s?1) at 20°C were also examined. Evaporative concentration of SM resulted in destabilization of casein micelles and dissociation of αS1- and β-casein, rendering CSM prone to further reactions. Treatment at 10 MPa and 1,000 s?1 at 20°C caused substantial dissociation of αS1- and β-casein in SM and CSM, with some dissociated caseins forming shear-induced soluble aggregates in CSM. The pressure applied at 10 MPa induced compression of the micelles and their dissociation in SM and CSM at 75 or 95°C, resulting in reduction of the micelle size. However, 10 MPa did not alter the mineral balance or whey proteins denaturation largely, except by reduction of some β-sheets and α-helices, due to heat-induced conformational changes at 75 and 95°C. 相似文献
17.
Soottawat Benjakul Friedrich Bauer 《Journal of the science of food and agriculture》2000,80(8):1143-1150
The effects of freeze–thaw cycles on the physicochemical and enzymatic changes of cod muscle proteins were investigated. The activities of α‐glucosidase and β‐N‐acetyl‐glucosaminidase increased as the the number of freeze–thaw cycles increased. A loss of Ca2+‐ATPase and Mg2+‐Ca2+‐ATPase activities was observed, while Mg2+‐EGTA‐ATPase activity increased, especially after one cycle of freezing–thawing, with a concomitant decrease in Ca2+ sensitivity. The surface sulphhydryl group content decreased with increasing freeze–thaw cycles, while no changes in total sulphhydryl group content were found. The surface hydrophobicity of actomyosin did not change significantly with the number of freeze–thaw cycles. The loss of protein solubility increased with increasing freeze–thaw cycles. However, no cross‐linked proteins induced by formaldehyde were found. The results revealed that freeze–thaw cycles directly affected the physicochemical and enzymatic properties of cod muscle proteins. © 2000 Society of Chemical Industry 相似文献
18.
Suw Young Ly 《Journal of the science of food and agriculture》2008,88(7):1272-1276
BACKGROUND: A voltammetric study of vitamin E (DL‐ α‐tocopherol) detection using square wave stripping and cyclic voltammetry is discussed in this paper. The working sensor was made by mixing carbon nanotube powder with DNA (double‐stranded calf thymus DNA) and mineral oil. In this electrode, the anodic peak was obtained for ? 0.6 V in a 0.1 mol L?1 phosphate electrolyte solution. RESULTS: Under optimized stripping conditions, analytical linear working ranges of 0.5–4.0 µg L?1 and 40.0–160.0 µg L?1 were obtained. The RSD precision was pegged at 0.105% with seven points using an 80 µg L?1 spike. The detection limit (S/N) was found to be 0.056 µg L?1 (1.30 × 10?10 mol L?1). CONCLUSION: The developed method was found to be applicable to quality control analysis in the food, pharmaceutical and other manufacturing sectors. Copyright © 2008 Society of Chemical Industry 相似文献
19.
Effect of phenolic acids on the rheological properties and proteins of hard wheat flour dough and bread 总被引:1,自引:0,他引:1
BACKGROUND: The effects of different phenolic acids on the rheological properties and gluten proteins of hard wheat flour dough and bread were investigated. Caffeic, ferulic, syringic and gallic acids were each blended with hard wheat flour at a concentration of 4.44 µmol L?1 g?1 flour. RESULTS: Mixing time and tolerance were reduced with the addition of phenolic acids. The phenolic acids reduced the maximum resistance to extension (Rmax) and increased the extensibility of dough, with effects in the following order: gallic < syringic < ferulic < caffeic acid. The effect on Rmax was more pronounced in overmixed dough. Loaf volume was most significantly decreased with the addition of caffeic acid. Extraction of sodium dodecyl sulfate‐soluble high‐molecular‐weight proteins was increased in both mixed and fermented doughs by the addition of ferulic and caffeic acids. The order of influence of the phenolic acids on the rheological properties and protein structure of dough and bread was consistent with that of their antioxidant activity. CONCLUSION: The addition of caffeic and ferulic acids reduced Rmax and increased the extensibility of hard wheat flour dough by modifying the high‐molecular‐weight gluten, which resulted in decreased bread volume. Copyright © 2011 Society of Chemical Industry 相似文献
20.
Linda Indrawati Richard L Stroshine Ganesan Narsimhan 《Journal of the science of food and agriculture》2007,87(12):2207-2216
Low‐field nuclear magnetic resonance (NMR) spin–spin relaxation (T2) measurements were used to study the denaturation and aggregation of β‐lactoglobulin (β‐LG) solutions of varying concentrations (1–80 g L?1) as they were heated at temperatures ranging from ambient up to 90 °C. For concentrations of 1–10 g L?1, the T2 of β‐LG solutions did not change, even after heating to 90 °C. A decrease in T2 was only observed when solutions having higher concentrations (20–80 g L?1) were heated. Circular dichroism (CD) spectroscopy and fluorescence tests using the dye 1‐anilino‐8‐naphthalene sulfonate (ANS) on 0.2 and 1 g L?1 solutions, respectively, indicated there were changes in the protein's secondary and tertiary conformations when the β‐LG solutions reached 70 °C and above. In addition, dynamic light scattering (DLS) showed that protein aggregation occurred only at concentrations above 10 g L?1 and for heating at 70 °C and above. The hydrodynamic radius increased as T2 decreased. When excess 2‐mercaptoethanol was added, the changes in both T2 and the hydrodynamic radius followed the same trend for all β‐LG protein concentrations between 1 and 40 g L?1. These observations led to the conclusion that the changes in T2 were due to protein aggregation, not protein unfolding. Copyright © 2007 Society of Chemical Industry 相似文献