纳豆芽孢杆菌发酵豆渣营养成分变化研究

研究纳豆芽孢杆菌发酵豆渣后营养成分变化情况。将湿豆渣调整水分,灭菌,冷却后接入不同浓度的纳豆芽孢杆菌,37℃下发酵22 h,4℃下后熟22 h后,对其粒径分布、营养成分、氨基酸含量、蛋白降解程度、部分抗营养因子指标进行测定。结果表明:豆渣经纳豆芽孢杆菌发酵后粒径变小、渣感减弱、口感明显得到改善。粗蛋白、粗纤维等常规营养成分无显著变化;水溶性蛋白、水溶性纤维、多糖、小肽含量显著提高(P0.05);氨基酸组分上,豆渣发酵后缬氨酸、甲硫氨酸等必需氨基酸含量均有显著提高(P0.05);豆渣风味品质得到明显改善;胰蛋白酶抑制剂活性和植酸含量均有显著下降(P0.05)。豆渣经纳豆芽孢杆菌发酵后营养价值和风味品质得到一定程度的提高,同时降低抗营养因子,为纳豆杆菌发酵豆渣的可行性提供理论参考。     

纳豆芽孢杆菌蛋白酶对大豆分离蛋白凝胶性的影响

利用纳豆芽孢杆菌发酵所产微生物蛋白酶对大豆分离蛋白进行酶法改性,以提高其凝胶性,并通过测定凝胶体系的流变学性质、水解程度和分析电泳图谱来探究纳豆芽孢杆菌蛋白酶对大豆分离蛋白凝胶性的影响.结果表明,体系凝胶强度受酶浓度和酶解温度的影响,酶浓度为1.072U/mL在40℃条件下形成的凝胶最稳定;同时凝胶强度与水解度有关,50℃时水解度超过12％,则不能形成稳定的凝胶.经微生物蛋白酶作用后,大豆分离蛋白的7s和11s均发生不同程度的水解.     

饼干模型对小麦及花生过敏原消化稳定性和免疫活性的影响

食品加工或食物基质可以不同程度地影响过敏原消化稳定性和免疫原性。然而,对食品加工和食物基质对 食物模型中过敏原的影响却知之甚少。本实验通过体外模拟胃肠消化的方式,包括模拟口腔咀嚼、胃部消化和十二 指肠消化,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹的方法,分析焙烤模型饼干中小麦过敏原和花生 过敏原的消化特性和免疫原性。结果显示：小麦和花生蛋白均可被胃蛋白酶迅速水解,醇溶蛋白、谷蛋白等致敏原 被降解成低分子质量多肽;可溶性蛋白中花生过敏原Ara h 1和Ara h 3基本消失,Ara h 2/6耐受胃肠消化;酶联免疫 吸附测定结果显示,消化后饼干中过敏原的致敏性降低。综合以上结果表明,饼干模型的消化性质基本不受焙烤加 工和其他基质的影响,免疫原性因致敏原被消化而降低。     

不同微生物液态发酵对蚕豆蛋白营养价值及功能特性的影响

该试验采用3种菌种(植物乳杆菌、黑曲霉、枯草芽孢杆菌)对脱脂蚕豆粉进行液态发酵,以蚕豆粉发酵前后的粗蛋白、酸溶蛋白、蛋白酶活力、氨基酸含量、持水性、持油性、起泡性等作为考察指标,对各菌种液态发酵蚕豆粉的营养价值及功能特性进行综合性评价。结果表明,经不同微生物液态发酵后,蚕豆粉多肽得率、酸溶蛋白、氨基酸含量均明显增加,经植物乳杆菌发酵后,多肽得率达到1. 46%,与未处理的蚕豆粉相比,其增幅高达71. 65%,效果极其明显;此外,经发酵后其多肽上清液抗氧化活性显著提升,在蛋白质量浓度为168μg/mL时,植物乳杆菌对DPPH的清除率达85. 46%,黑曲霉对ABTS自由基的清除率达87. 89%,经微生物液态发酵后蚕豆的营养价值得以大幅度提升; 3种菌种液态发酵有助于改善蚕豆蛋白的功能特性,植物乳杆菌的作用效果较优。     

不同菌种发酵对辣木叶蛋白降解的影响

以辣木叶粉为原料,研究自然发酵和7株不同菌株接种发酵过程中理化指标（pH、总酸、蛋白酶活力、蛋白含量、氨基酸态氮含量）的变化规律,并采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）筛选降解辣木叶蛋白的优良菌株。结果表明,发酵过程中pH值随发酵时间延长而降低,总酸含量逐渐上升,蛋白酶活力呈先上升后下降趋势。7株不同菌株中,纳豆芽孢杆菌（Bacillus natto）和棉籽糖乳球菌（Lactococcus raffinolactis）发酵的辣木叶粉在24 h时蛋白酶活力最高,分别为270.58 U/mL和248.34 U/mL,发酵结束时,蛋白含量分别下降20.24 mg/g、21.95 mg/g,氨基酸态氮含量明显高于其他组。SDS-PAGE结果显示,纳豆芽孢杆菌和棉籽糖乳球菌能够降解分子质量70 kDa和35 kDa的两种主要蛋白。因此该研究选出纳豆芽孢杆菌和棉籽糖乳球菌作为发酵降解辣木蛋白的优良菌株。     

发酵酶解法提取紫菜蛋白多肽及其特性研究

利用纳豆芽孢杆菌或枯草芽孢杆菌对紫菜进行发酵,并结合木瓜蛋白酶酶解的方法提取紫菜蛋白多肽,考察了发酵酶解法对蛋白提取率的影响。结果发现,以未加热处理的紫菜为原料,纳豆芽孢杆菌接种量4%,在30℃下发酵24 h,再利用木瓜蛋白酶(1 mg/mL)酶解3 h后,紫菜蛋白提取率为82%,而利用接种量2%的枯草芽孢杆菌时蛋白提取率可达89%。根据紫菜发酵酶解物的性质分析结果,发现提取物的氨基酸组成与紫菜相同,蛋白溶解性不受pH影响,在pH 3.0下乳化活性最高,而在pH 9.0下乳化稳定性最好。     

酶解时间、分子量分布对于花生致敏蛋白含量的影响

花生是我国一种重要的粮食作物,但是同时花生蛋白也是重要的食物过敏原。目前国内关于花生蛋白改性后致敏活性与蛋白分子量分布、水解程度的研究报道相对较少。采用胰蛋白酶和木瓜蛋白酶对花生蛋白粉进行限制性水解,通过超滤手段逐级分离蛋白水解物,酶联免疫结果显示:随着水解时间的增加,致敏蛋白含量逐渐增加,未水解的蛋白提取物致敏蛋白含量为71.19 mg/g蛋白,水解4 h后,致敏蛋白含量可达356.74 mg/g;超滤后,致敏蛋白含量显著降低,最低仅为1.91 mg/g,与4 h后的水解液相比较致敏蛋白含量降低了180倍。综上,酶解结合超滤技术是降低花生致敏蛋白的有效方法,可以用于开发低过敏的花生功能食品。     

复合菌种发酵对蚕豆蛋白理化性质及其功能特性的影响

该实验将植物乳杆菌(ZR)、嗜热链球菌(SL)和嗜酸乳杆菌(SS)3种菌种两两复配及三菌复合作为发酵剂用于脱脂蚕豆粉的液态发酵，测定发酵前后蚕豆粉蛋白酶活性、肽含量、可溶性蛋白含量、吸水性和吸油性、持水性和持油性、起泡和泡沫稳定性、凝胶特性和蛋白质溶液的黏度等指标，评价每种复合物液体发酵后脱脂蚕豆粉的营养价值和功能特性。实验结果表明，在发酵条件一致的情况下，ZR+SL+SS联合发酵后，发酵液蛋白酶活性最高、多肽含量达到30.94%,脱脂蚕豆粉可溶性蛋白含量比未发酵时提高3.23倍(P<0.05)。ZR+SS复合发酵后，脱脂蚕豆粉可溶性蛋白含量达到98.86%、蛋白起泡性达到32.00%、泡沫稳定性达13.33%,均高于其他组(P<0.05)。经4种复合菌发酵后蛋白质硬度、弹力、吸水(油)性等明显增加，弹性下降。此外除经过SL+SS复合菌发酵的脱脂蚕豆粉蛋白溶液黏度降低之外，其余组增加，其中经ZR+SS复合发酵后黏度达到55.20 mPa·s。4种复合菌发酵后蛋白营养价值均有了显著提高，表明复合菌种发酵有助于改善蚕豆蛋白的功能特性。     

微生物混合发酵提高麸皮营养价值的研究

以普通麸皮为原料,选取黑曲霉、纳豆芽孢杆菌、酿酒酵母和粪链球菌进行混菌固态发酵,研究其对麸皮营养价值的提高.结果表明,最适装料量为1/1;最适料水比为1.0∶0.8;接种比例黑曲霉∶纳豆芽孢杆菌∶酿酒酵母∶粪链球菌=0.5∶2∶10∶4.存37℃发酵48h,发酵麸皮中有益菌总数达到109个/g干物料,粗蛋白含量(绝干)由16.1%提高为20.97%.粗纤维含量由10.5%降低为5.32%,产品有很浓郁的酸香味.     

枯草芽孢杆菌发酵制备花生多肽及其自由基清除活力的研究

为优化枯草芽孢杆菌发酵制备花生多肽的条件,采用响应面法,以水解度为响应值,研究了pH、接种量、发酵时间和发酵温度对制备高水解度花生多肽的影响.最终确定最佳发酵条件为:pH7.2,接种量2％,发酵时间3.97 d,发酵温度32.6℃.在此条件下发酵制得的花生多肽水解度为22.42％,与理论预测值的误差在1％以内,说明利用建立的模型在实践中进行预测是可行的.并对制备的花生多肽进行自由基清除试验,发现该条件下制备的花生多肽具有较好的DPPH自由基清除能力和超氧阴离子自由基清除能力.     

质谱法分析烘焙对花生过敏原Ara h 1潜在致敏性的影响

为研究烘焙对花生过敏原Ara h 1潜在致敏性的影响,采用高离液序列盐溶液从鲜花生和烘焙花生中提取总蛋白,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析烘焙前后蛋白条带变化情况,并对其中的花生主要过敏蛋白Ara h 1条带进行质谱和Swiss-Model模型分析.十二烷基硫酸钠-聚丙烯酰胺凝胶电泳结果显示,烘焙花生蛋白出现...     

Allergenicity of roasted peanuts treated with a non-human digestive protease

Peanut allergy is a severe and lifelong type of food allergy triggered by allergenic proteins and peptides in peanuts. This study investigated the effects of ultrasound-assisted alcalase treatment on the concentrations of major allergenic proteins (Ara h 1 and Ara h 2) in roasted peanut kernels and the allergenicity of treated peanut extracts. Peanut kernels were sonicated for 1 h in buffer solution, incubated with different amount of alcalase for various time, then vacuum dried. The variations of Ara h 1 and Ara h 2 contents in soluble and insoluble portions of peanuts treatments were evaluated by sandwich ELISA and SDS-PAGE, respectively. The in vitro IgE-binding capacity of treated peanut extracts was determined by a competitive inhibition ELISA using pooled plasma of 10 peanut allergic patients. Samples with lower in vitro IgE-binding were used for human skin prick tests (SPTs) in peanut allergic individuals. Results indicate that alcalase digestion of sonicated peanuts significantly increased protein solubility while decreasing Ara h 1 and Ara h 2 concentrations in both soluble and insoluble portions of peanuts relative to untreated peanuts. The maximum reductions of Ara h 1 and Ara h 2 levels were obtained following 3 hour digestion with alcalase at concentrations of 4.54 and 6.05 U/100 g. Samples obtained under these conditions showed the lowest in vitro IgE-binding and caused the least allergic response in human SPTs. The current study suggests that the allergenic potential of peanuts could be reduced by postharvest processing such as ultrasound-assisted enzymatic treatment of peanuts kernels.     

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.     

加工方式和蛋白提取方法对花生蛋白致敏性的影响

加工方式影响花生蛋白的致敏性。本实验系统的研究了中国传统花生制品(水煮和油炸花生)和美国常食用的烘烤花生致敏性的差异,并比较了不同蛋白提取缓冲溶液对花生致敏蛋白提取效率的影响。结果表明:不同花生制品在不同的缓冲溶液中蛋白提取效率不同,水煮和油炸并没有较烘烤方式降低致敏蛋白的数量,而降低花生蛋白的致敏性。因此加工方式并不是导致中国花生过敏发病率低的主要原因。     

Beneficial Influence of Short‐Term Germination on Decreasing Allergenicity of Peanut Proteins

Most allergenic storage proteins in peanuts are degraded during seed germination. By altering this natural physiological process, it might be possible to reduce peanut protein allergenicity. However, little is known about the change in allergenic proteins and their corresponding immunocreactivity, and the effects of major environmental conditions on their allergenicity during germination. In this study, the influence of different germination conditions (temperature and light) on the degradation of Ara h1 and allergenicity changes of peanut seeds was evaluated by ELISA and Western blotting. The results showed that the 40‐ and 65‐kDa proteins in peanut seeds degraded rapidly during the time course, beginning at 60 (at 25 °C) and 108 h (at 20 °C), and the corresponding immunocreactivity of Ara h1 decreased approximately one‐third after 5 to 7 d of germination. Compared with the cotyledons, the embryonic axes had a higher proportion of Ara h1, which was then degraded relatively faster during germination, resulting in a significant reduction in its allergenicity. Although a higher temperature improved the seed germination rate, it affected sprout quality (as did light); therefore, 25 °C and dark surroundings were suitable conditions under which peanut sprouts were processed; neither factor significantly affected the allergenicity of Ara h1. These results provided a theoretical basis for studies using biological methods to reduce peanut allergenicity.     

Peanut Allergy: Characteristics and Approaches for Mitigation

Peanut allergy has garnered significant attention because of the high sensitization rate, increase in allergy, and severity of the reaction. Sufficiently reliable therapies and efficient mitigating techniques to combat peanut allergy are still lacking. Current management relies on avoiding peanuts and nuts and seeds with homologous proteins, although adverse events mostly occur with accidental ingestion. There is a need for hypoallergenic peanut products to protect sensitized individuals and perhaps serve as immunotherapeutic products. Alongside traditional practices of thermal and chemical treatment, novel processing approaches such as high‐pressure processing, pulsed ultraviolet light, high‐intensity ultrasound, irradiation, and pulsed electric field have been performed toward reducing the immunoreactivity of peanut. Covalent and noncovalent chemical modifications to proteins also have the tendency to alter peanut allergenicity. Enzymatic hydrolysis seems to be the most advantageous technique in diminishing the allergenic potential of peanut. Furthermore, the combined processing approach (hurdle technologies) such as enzymatic hydrolysis followed by, or in conjunction with, roasting, high pressure and heat, ultrasound with enzymatic treatment, or germination have shown a significant reduction of peanut immunoreactivity and may emerge as useful techniques in reducing the allergenicity of peanut and other foods. This study represents our current knowledge about the alterations in allergenic properties of peanut via different processing mechanisms as well as evaluating its future potential, geographical based data on increasing sensitization, clinical relevance, eliciting dose, and current management of peanut allergy. Furthermore, the molecular characteristics and clinical relevance of peanut allergens have been discussed.     

双酶分步水解制备花生多肽工艺优化

为了开发和利用花生蛋白资源,生产高附加值蛋白产品,以花生分离蛋白为原料,采用Alcalase 和Flavourzyme 分步水解法制备花生多肽。通过单因素试验和响应面中心组合设计试验,研究Flavourzyme 水解花生分离蛋白过程中加酶量、底物质量分数、酶解温度、酶解时间和酶液pH 值等因素对水解的影响。建立水解液中可溶性氮质量浓度与各种影响因素的回归模型;确定Flavourzyme 酶解反应的最佳工艺参数为pH7.0、加酶量1714U/g 底物、底物质量分数5%、酶解温度55℃、酶解时间90min。在此条件下,酶解产物中可溶性氮质量浓度为19.44mg/mL。     

混菌固态发酵花生粕的工艺优化

采用纳豆芽孢杆菌和红曲霉混合菌株固态发酵花生粕,以纳豆激酶的活力和γ-氨基丁酸的含量为指标,通过单因素实验和正交试验对发酵的温度、时间、料水比、菌种比例、接种量进行优化,确定最佳工艺参数为:温度31℃,发酵时间46 h,料水比为1:0.4 g/mL,菌种比例(纳豆芽孢杆菌:红曲霉)2:1,接种量6%,在此条件下测定纳豆激酶的活力为(844.56±13.80) U/g,γ-氨基丁酸含量为(105.25±0.25) mg/g,对羟自由基清除率为68.46%±0.16%,对DPPH·清除率为76.98%±0.95%,铁还原力的OD值为0.481。     

酶法水解花生蛋白及产物抗氧化活性的研究

本文以花生粕中提取的花生蛋白为原料,分别使用Alcalase、Protamex和Papain酶解,研究不同酶解时间酶解产物的水解度、三氯乙酸中可溶性氮含量和抗氧化活性的变化规律。结果表明:随着酶解时间的延长,Alcalase、Protamex和Papain酶解花生蛋白的酶解产物的水解度先逐渐增加后趋于稳定,三氯乙酸中可溶性氮含量先增加后减小,超氧阴离子自由基清除能力先增加后有小幅降低。不同酶的酶解产物的抗氧化性能与其水解能力并不直接相关。     

Use of Bacillus subtilis to enrich isoflavone aglycones in fermented natto

BACKGROUND: Natto is a food made by fermenting cooked soybeans with Bacillus subtilis. Soybean isoflavones are reported to provide many health benefits, including oestrogenic effects. However, isoflavone aglycones may be absorbed faster and in higher amounts in the human intestine than their glucosides. This study aimed to investigate the content of isoflavone components in commercial natto products as well as the use of B. subtilis strains to ferment cooked soybeans to produce a high level of isoflavone aglycones in natto. RESULTS: The content and composition of isoflavones in commercial natto products were predominantly (>76%) isoflavone glucosides. Fermentation of cooked soybeans with B. subtilis BCRC 14718 at 37 °C for 48 h was more effective in converting glucosides to aglycones than with other strains of B. subtilis, increasing the proportion of isoflavone aglycones from 12 to 68% of the total isoflavones in the fermented natto. The proportions of the isoflavone aglycones daidzein and genistein in cooked soybeans fermented with B. subtilis BCRC 14718 for 48 h increased from 6 to 54% and from 5 to 13% respectively. CONCLUSION: Bacillus subtilis BCRC 14718 incubated with cooked soybeans produces higher levels of isoflavone aglycones, which may enhance health benefits over traditional fermented natto. Copyright © 2008 Society of Chemical Industry