Heat denaturation of Brazil nut allergen Ber e 1 in relation to food processing

Ber e 1, a major allergen from Brazil nuts, is very stable to in vitro peptic digestion. As heat-induced denaturation may affect protein digestibility, the denaturation behaviour of Ber e 1 was investigated. The denaturation temperature of Ber e 1 varies from approximately 80–110 °C, depending on the pH. Upon heating above its denaturation temperature at pH 7.0, the protein partly forms insoluble aggregates and partly dissociates into its polypeptides, whereas heating at pH 5.0 does neither induce aggregation, nor dissociation of the protein. The denaturation temperature of approximately 110 °C at pH values corresponding to the general pH values of foods (pH 5–7) is very high and is expected to be even higher in Brazil nuts themselves. As a result, it is unlikely that heat processing causes the denaturation of all Ber e 1 present in food products. Consequently, the allergen is assumed to be consumed (mainly) in its native form, having a high stability towards pepsin digestion.     

毛叶枣贮藏保鲜技术研究

分别对毛叶枣的贮藏适性、涂膜保鲜技术及1-MCP对毛叶枣的贮藏性的影响进行了较深入地研究。试验结果表明:贮藏温度2～5℃,RH90%～100%范围为毛叶枣适宜的贮藏条件;经过1-MCP处理的毛叶枣果实贮藏性明显高于未经处理的果实。涂膜保鲜试验表明:以瓜尔豆胶1.1g、卡拉胶2.75g、分子蒸馏单甘酯0.5g、乙醇50mL、水500mL、杀菌剂和防腐剂若干配制的涂膜剂处理毛叶枣果实,在室温条件下其货架可达到12d,在2～5℃贮藏条件下其贮藏期可达50d,商品率100%。     

巴西坚果过敏原Ber e 1的分离纯化及表征鉴定

以巴西坚果仁为原料,通过粉碎、脱脂、浸提、阴离子交换层析纯化过敏原蛋白Ber e 1;利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、液相色谱-串联质谱联用、Western blot等方法对其进行鉴定,并通过圆二光谱仪和紫外分光光度计表征其二、三级结构。结果表明：纯化获得的巴西坚果过敏原Ber e 1,单轮制备量可达20 mg以上,且纯度大于95%,其蛋白质高级结构未被破坏,能够被巴西坚果过敏患者的血清准确识别。该纯化技术路线简单、对设备要求低且高效,为巴西坚果过敏原Ber e 1的相关研究奠定了一定的基础。     

Preparation of transparent pea protein gels: a comparison of isolation procedures

Two commercially viable procedures for the preparation of protein fractions from pea flour are described and discussed. These fractions form thermally induced transparent gels at acid pH and are composed of the globulins, vicilin and legumin, identified by electrophoresis and differential scanning calorimetry. The gels have clarities that allow their use as vegetarian substitutes for gelatin and additionally show thermal stabilities suitable for applications in hot products. The influence of other food ingredients (salt, sugar and vegetable oil) on gel clarity is also investigated.     

Peanut varieties with reduced Ara h 1 content indicating no reduced allergenicity

Peanut allergy is a major cause of food‐induced severe anaphylactic reactions. To date, no medical care is available to prevent and treat peanut allergy and therefore hypoallergenic peanut varieties are of considerable health political and economic interest. Major allergens that induce IgE‐responses in peanut‐sensitive patients are Ara h 1, Ara h 2 and Ara h 3/4. In order to identify hypoallergenic peanuts, commercially locally available peanut varieties were screened for their allergen content. Ara h 1‐deficient peanuts from Southeast Asia were identified by SDS‐PAGE, immunoblotting, inhibition assays and ELISA. 2‐D PAGE analyses demonstrated the different compositions of the tested extracts and revealed a number of variations of the allergen patterns of peanuts from different varieties. Mediator release experiments of these peanut extracts demonstrated similar allergenicities as compared with standard peanut extract. These results indicate that the allergenicity of peanuts with reduced Ara h 1 content might be compensated by the other allergens, and thus do not necessarily cause a reduction of allergenicity.     

In-vitro rates of rumen proteolysis of ribulose-1,5-bisphosphate carboxylase (rubisco) from lucerne leaves,and of ovalbumin,vicilin and sunflower albumin 8 storage proteins

Laboratory systems were developed, based upon in-vitro incubations with rumen fluid, to examine the rate of proteolysis (ie degradation) of sunflower albumin 8 (SFA 8; 24% sulphur amino acids; SAA) from sunflower seeds, ovalbumin (6% SAA) from chicken egg white, ribulose-1,5-bisphosphate carboxylase (Rubisco; 3% SAA) from lucerne leaves, and vicilin (0% SAA) from pea seeds. After fractionation by SDS-PAGE, proteins were analysed by either Western blotting, using specific antibodies (SFA 8, vicilin and ovalbumin) or by Coo-massie blue staining (Rubisco). Proteolysis of the large subunit of Rubisco occurred very quickly and as two components, with half-lives of 11.6 and 1.5 h. The small subunit of Rubisco was more resistant to degradation, with a half-life of 17.3 h. Vicilin was degraded extremely rapidly (half-life 0.16 h). SFA 8 and ovalbumin both showed resistance to degradation, but by two different mechanisms. Ovalbumin was not degraded at all during the initial 16 h of incubation, but then degraded with a half-life of 8.7 h. SFA 8 (mol wt 12 100) disappeared very rapidly, with a half-life of 3.0 h. The degradation of SFA 8 was associated with the appearance of a polypeptide (mol wt 8000), which was extremely resistant to degradation, with a half-life of 69.3 h. It was concluded that both the number of disulphide linkages and tertiary structure were important in determining resistance of proteins to rumen degradation, and that incorporation of SFA 8 and ovalbumin proteins into forages using genetic engineering techniques would be likely to increase the quantity of SAA by passing rumen fermentation.     

Heat Induced Gelation of Pea (Pisum sativum) Mixed Globulins, Vicilin and Legumin

Mixed globulins (MG) were extracted from ground dry peas (Pisum sativum, B-160) with 0.5M NaCl, 50 mM potassium phosphate, pH 7.2, and isolated by precipitation at pH 4.5. Crude vicilin and legumin were fractionated from the MG by dialysis against 0.2M NaCl, pH 4.8, and centrifugation, then further purified using DEAE-cellulose chromatography. Conditions for maximum gel hardness of heat induced MG gel, as determined with an Instron Universal Testing Machine, were heating for 20 min at pH 7.1 at 87°C. Purified vicilin, but not legumin, formed heat induced gels. The relationship was linear between protein (globulin) concentration and log gel hardness. At all protein concentrations studied, as proportion of legumin decreased, gel hardness increased.     

Isolation of conglutin δ, a sulphur-rich protein from the seeds of Lupinus angustifolius L.

Although a 2S globulin class has been observed in salt extracts from seeds of several lupin species, there have been conflicting reports regarding the importance of this class in Lupinus angustifolius. Conglutin δ, a major sulphur-rich protein extracted from mature seeds of L. angustifolius cv. Uniwhite, was separated by gel-filtration into two oligomeric forms. The sedimentation coefficients of conglutin δ₁ (20%) and conglutin δ₂ (80%), were 3.2S and 2.0S respectively. The amino acid compositions of both oligomeric forms of conglutin δ were identical and similar to the compositions published for the 2S globulins in other lupin species. Because of the low level of tyrosine and the absence of tryptophan, conglutins δ₁ and δ₂ showed little absorbance at 280nm (E^1%/1 cm^?23). They were characterised by unusually high levels of glutamic acid and 1/2 cysteine (38.5 and 8.5 residues percent respectively) while methionine was absent. Gradient SDS-PAGE showed that conglutins δ₁ and δ₂ were homogeneous single-subunit species. On reduction, with or without S-carboxymethylation, both the conglutin δ₁ and δ₂ subunits yielded similar pairs of large and small polypeptide chains. Since conglutin δ rarely resolves from conglutin a during electrophoresis on cellulose acetate membranes in phosphate buffer at neutral pH, this method is not as useful for screening lupin cultivars as has been claimed previously.