CN1491085A - 含大豆蛋白和抑制素的食物制品 - Google Patents
含大豆蛋白和抑制素的食物制品 Download PDFInfo
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- CN1491085A CN1491085A CNA028048083A CN02804808A CN1491085A CN 1491085 A CN1491085 A CN 1491085A CN A028048083 A CNA028048083 A CN A028048083A CN 02804808 A CN02804808 A CN 02804808A CN 1491085 A CN1491085 A CN 1491085A
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- food article
- inhibin
- food
- soybean
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Abstract
本发明描述了一种适合用于降低低密度脂蛋白胆固醇含量的食物制品,该食物制品的每平均一份中含有至少5克大豆蛋白和至少5mg/kg的抑制素。优选,该食物制品中含有大豆发酵成分。
Description
技术领域
本发明涉及适用于降低人血液中低密度脂蛋白胆固醇含量的食物制品。
发明背景
心血管疾病是导致发病和死亡的一个原因,特别是在美国和西欧国家,并且在发展中国家正在显露。据提及,与心血管疾病发育有关的几个因素包括疾病的遗传因素、性别、生活方式诸如吸烟和饮食、年龄、高血压以及高脂血包括血胆固醇过多。这些因素中的一些,特别是高脂血和血胆固醇过多,有助于动脉粥样硬化的发育,是血管和心脏疾病的首要原因。
低密度脂蛋白胆固醇(以下称“LDL-胆固醇”)升高与增加患冠心病的危险有直接关系。
人所食用的食品中存在的大豆蛋白,可以降低低密度脂蛋白胆固醇(LDL-胆固醇)的含量和降低冠心病的危险。
含有大豆蛋白的食物制品是公知的。可能含有大豆蛋白的食物制品的实例是肉制品、焙烤制品、食品模拟物和乳制品。Waggle,D.H.和Kolar,C.W.,“大豆蛋白和人的营养(Soy protein and humannutrition)”,proceedings of a symposium held May 222-25,1978,Keystone Colourado,第19-51页中描述了大豆蛋白的类型和大豆蛋白的食品用途。
大豆蛋白物料已知可以降低动物血液中总胆固醇和LDL-胆固醇含量。大豆蛋白摄取量对人血清脂质的效果分析显示,饮食中的大豆蛋白与降低人的总胆固醇和LDL-胆固醇血清浓度有显著的关系(Anderson,Johnstone和Cook-Newell,N.Engl.J.Med.,Vol.333,No.5,pp.276-82(1995))。
抑制素是已知对人血液中LDL-胆固醇含量具有降低效果的化合物。抑制素可抑制羟甲戊二酸单酰辅酶A(HMG-CoA)还原酶,胆固醇生物合成中的速控步骤。
科学家研究证实了抑制素特别针对LDL血液胆固醇和三酸甘油酯含量降低活性方面的健康特性,并且在动物和人中都有这种特性(Li等,Nutrition Research 18,71-81(1998);Heber等,Am.J.Clin.Nutr.69,231-236(1999))。
关于抑制素效果的早期报告是在1979年作出的。日本科学家Endo从红曲霉中分离出可人工降低大鼠中诱导型高脂蛋白血的代谢物(Endo,J.Antibiotics 32,852-854,(1979))。这些代谢物已知是红曲霉素。红曲霉素等同于胆固醇降低药物洛伐他丁。洛伐他丁由Merck公司以商品名称Mevacor出售。洛伐他丁的衍生物,辛伐他丁,作为胆固醇降低药品以名称Zocor出售。洛伐他丁的其它衍生物,例如普伐他丁和美伐他丁,也作为血胆固醇过多的液体降低药品出售。在日本,红曲霉-提取物以胶囊的形式作为饮食制品红曲霉素出售。上述抑制素的通常剂量为20mg/天,具有至少20%血液LDL-胆固醇降低的效果。
据报导,除上述红曲霉菌种外,使用其它真菌的发酵中也能产生抑制素。据显示,抑制素可以通过各种丝状真菌来生产,包括红曲霉、曲霉属、青霉属、灰侧耳菌属、腐霉属、隐壳霉属、拟青霉属、Eupenicillium和束孢霉属。
药物制剂中所用的抑制素的制备和纯化包括很多步骤,其中所用的成分都是食品工业中不常用的。相比具有较少加工步骤的工艺,很多加工步骤增加了成本。出于这些原因,为药用而制备的抑制素是无法在食品工业中得到使用的。
作为食物制品,用红色红曲霉属真菌发酵的稻米(红米)是已知的,并且在中国已有上百年的历史。红米得到使用并且仍在造酒中使用,作为食品着色剂和作为传统中国医药中的药品。我们发现,市场上可获得的大部分红米中并不含抑制素或含非常少量的抑制素。食品和药品管理局(The Food and Drug Administration)的结论认为,可从市场上获得的红色酵母米中不含显著量的洛伐他丁(FDA,Docket No.97-0441,Final Decision)。
WO 99/23996描述了一种用于治疗高血清胆固醇和/或三酸甘油酯的含有红米制品的组合物,其中所说的红米制品中含有至少0.05wt%洛伐他丁。
红米粉末胶囊由Pharmanex公司作为饮食添加物的形式以名称Cholestin出售。Pharmanex还销售含有红色酵母米(Monascus purperuswent)的Cholestin条状食品。
红米具有强烈的红色。红米的深红色当用作着色剂时是有利的,而当在大多数食物制品中使用时是不利的。由于红米制品具有强烈红色,由红米制备的食品,根据向食物制品中添加的红米制品的量,而被染成黄色、橙色或红色。食品中添加的红米的量越高,食物制品的红色就越深。在已知的食物制品中,为添加足够的抑制素就不得不添加相对大量的红米。这样就不可避免地导致制品带有红颜色。
在一些食物制品中,红米染色是不期望的。特别是在西方世界,消费者不愿意使用那些他们所习惯的颜色被改变了的产品。例如涂抹料,包括人造奶油、奶油、低脂涂抹料和烹调油或色拉油,当这些产品的颜色是深黄色或橙色或红色时,消费者认为是不可接受的。然而,同时申请人发现,这些类型的产品却是每日摄取足够量抑制素以获得血液LDL-胆固醇降低效果的杰出载体。
发明概述
本发明的一个目的是提供一种食物制品,这种食物制品没有因添加抑制素而产生的不期望的颜色。本发明的另一个目的是增加含抑制素的已知食物制品的保健效果。本发明的再一个目的是提供一种含抑制素的食物制品的制备方法,该方法中包括的加工步骤少于制备药物形式的抑制素时的步骤。另一种目的是提供一种方法,该方法可避免使用食品工业中不常用的配料或加工助剂。这些目的的一个或多个由本发明来实现。
我们目前发现了一种每平均份中含有至少5克大豆蛋白的食物制品,其特征在于该食物制品中含有至少5mg/kg的抑制素。
有利的是,食物制品中含有大豆发酵成分。
意想不到地,我们发现当用于红曲霉发酵的底物是大豆和/或大豆成分时,可以避免红米发酵中的发酵产物变成红色,即可获得不变色或仅微微变色的发酵产物。此外,我们发现存在于大豆中的具有有益健康效果的化合物也存在于该发酵产物中。这些化合物包括(但不限于)多不饱和脂肪酸、植物甾醇、膳食纤维包括可溶性纤维、多酚和皂苷。发酵产物中含有这些化合物的结果是使本发明的食物制品,相比含有抑制素的已知食物制品,其保健效果得到增加。
发明详述
本文中将使用如下定义。
抑制素定义为具有下式(I)所示结构式的物质。
在此结构式中,R1和R2可以是任何基团。优选的抑制素是下表1中给出的化合物。
表1:根据式(1)的优选的抑制素
R1 R2
红曲霉素K CH3
红曲霉素L CH3 H
红曲霉素J CH3 OH
红曲霉素X CH3
密实菌素 H
(ML-236B)
ML-236-A H OH
NL-236-C H H
本文中的多酚是指植物来源的多酚。包括黄酮类化合物,其中包括异黄酮。多酚包括异黄酮、1,2-二苯乙烯、木酚素、香豆雌醇和resorcyclicacid内酯。异黄酮的实例是染料木素、大豆黄酮、牛尿酚、大豆黄素、鸡豆黄素A、香豆雌酚、maitaresinol、芒柄花黄素、O-去甲基engolesin、肠内酯和肠二醇。本发明中优选的异黄酮是染料木素和大豆黄酮以及大豆黄素,它们存在于大豆中。
本文中,皂苷是以β-D-吡喃葡糖苷醛酸衍生物的形式获得。皂苷的实例是大豆三萜苷元(soya sapogenol)A、B、C、D和E,大豆皂苷元I、II和III,如Lebensmittel Lexikon,B.Behr′s Verlag GmbH &Co.Hamburg,Bd.2,L-Z-3,1993,第550-552页中所述。
多不饱和脂肪酸酯定义为在脂肪酸链中具有一个以上不饱和基团的脂肪酸酯。多不饱和脂肪酸酯的实例是亚油酸酯、亚麻酸酯、花生四烯酸酯。
本文中,膳食纤维是指对人的胃肠酶消化具有耐受性的各种植物物质的集合性术语。取决于它们的溶解性,膳食纤维可以分成不溶性的(纤维素,一些半纤维素,木质素)和可溶性的(其余的半纤维素,树胶,粘胶)。大豆colyledon纤维中同时含有可溶性和不溶性的膳食纤维。
本文中,植物甾醇定义为由植物产生的甾醇化合物,其结构上非常类似于胆固醇,不同之处是它们在甾醇侧链上的C24位含有一些取代基。植物甾醇包括4-去甲基甾醇、4-单甲基甾醇、4,4′-二甲基甾醇及其混合物。这种植物甾醇的实例是β-谷甾醇、菜油甾醇、豆甾醇。本文中,术语“植物甾醇”还包括饱和植物甾醇(phytostanols),植物甾醇的饱和等价物。
多酚、多不饱和脂肪酸、植物甾醇、蛋白质、肽、膳食纤维和皂苷在下面将统称为大豆活性物。
除非另有说明,所给出的量按wt%或每百万分之重量份(ppm),mg/kg或g/kg计,相对于食物制品的总重量(除非另有说明)。
除非另有说明,本文中给出的抑制素的量是每种抑制素的量的总和,例如通过色谱法测定的。
本文中,底物定义为:当使用水基发酵培养基时,发酵培养基中溶剂除外、例如水除外的总的化合物。当不含溶剂时,底物等于发酵培养基。
本文中给出的蛋白质量是每种蛋白质的量的总和,除非有另外的说明。
大豆活性物的量按多不饱和脂肪酸、植物甾醇、蛋白质、肽、膳食纤维(包括可溶性纤维)、多酚和皂苷的总和(wt%或ppm)计。
本发明的食物制品优选是其中将大豆蛋白物料用作功能性成分的食品。包括(但不限于):肉类,诸如肉馅、乳化肉、发酵肉和腌制肉;饮料类,诸如营养饮料、运动饮料、蛋白质强化饮料、果汁、乳、乳替代品和减重饮料;干酪和类似干酪的产品,诸如豆腐;冷冻甜食,诸如冰淇淋、冰乳、低脂肪冷冻甜食和非乳制品制冷冻甜食、酸奶;汤料;调味汁,诸如酱油;布丁;早餐谷物制食品;面食制品;焙烤制品,诸如面包和蛋糕;色拉饰料以及浸泡料和涂抹料,诸如蛋黄酱、加碎屑的浸泡料、低脂肪涂抹料、夹心面包涂抹酱;疗效制品,例如减肥制品或代餐品等等。
优选,本发明的食物制品不包括特别是适合用于饲养动物的制品(饲料)。
如果将此食物制品按照消费者的普通需要使用时,食物制品中存在的抑制素和大豆蛋白的量应当足以获得血液LDL-胆固醇降低效果。
本领域技术人员能够调节抑制素和大豆蛋白在食物制品中的百分比来达到上述效果。此百分比取决于食物制品的类型,因为使用一份食物制品的量值是不同的。此外,食物制品的消费模式(每天一份和几天内分配)取决于食物制品。关于一份食物的量值数据可见美国食品和药品管理局(the United States Food and Drug Administration)(FDA)出版的标题为“每份食物常规消费的参考量(Reference Amountscustomarily consumed per eating occasion)”的目录表。
作为示例,下表2中显示了一些本发明可以制备的制品以及典型的一份的量。
表2
制品 一份的平均量(g)
人造奶油 15
肉制品 50
条状食品 60
代餐饮料 330
酸奶型饮料 200
冰淇淋 75
谷物制食品 50
饮料 200
优选,本发明的食物制品中含有非糖基化的异黄酮。在大豆以及从大豆获得的大豆物料中,异黄酮基本上是以糖基化的形式存在。一般来说,约5wt%的异黄酮是以非糖基化的形式存在的。最重要的糖基化异黄酮是染料木苷,大豆苷和glycetin。非糖基化的形式分别是染料木素、大豆黄酮和甘油黄酮(glycetein)。染料木素、大豆黄酮和甘油黄酮(glycetein)据报道具有有利于健康的效果,包括雌激素和抗氧化剂特性。
我们发现,由于本发明的发酵使糖基化的异黄酮转变成相应的非糖基化的异黄酮,从而达到更有益。例如,相比未发酵的大豆,在发酵大豆中的染料木素和大豆黄酮的量有所增加。意想不到的是,随着这种有利转化的同时还产生出抑制素。
因此,本发明还涉及一种含有抑制素、大豆蛋白、染料木素和染料木苷的食物制品,其中抑制素的量为5-500mg/kg,大豆蛋白的量为10-500g/kg并且染料木素的量为染料木素和染料木苷总量的10-99wt%、优选15-99wt%、更优选20-95wt%、进一步优选20-90wt%、最优选20-80wt%。
本发明还涉及一种含有抑制素、大豆蛋白、大豆黄酮和大豆苷的食物制品,其中抑制素的量为5-500mg/kg,大豆蛋白的量为10-500g/kg并且大豆黄酮的量为大豆黄酮和大豆苷总量的10-99wt%、优选15-99wt%、更优选20-95wt%、进一步优选20-90wt%、最优选20-80wt%。
对每种食物制品而言,染料木素和大豆黄酮的绝对量可以由本领域技术人员来调整至需要的水平。这可以例如通过从具有不同异黄酮含量的物料中选择发酵用的大豆物料;通过调节发酵条件,诸如发酵时间以及通过选择向食物制品中添加的发酵大豆的量来实现。按此方式,可以将所说的量调整至致使每天摄取需要的异黄酮,例如对染料木素而言可以是摄取50-80mg/天。本发明食物制品中染料木素的优选的绝对含量,取决于食品的类型,并且可以是50mg/kg或更多、更优选100mg/kg或更多、200mg/kg或更多、500mg/kg或更多,最优选200-5000mg/kg。同样,大豆黄酮的绝对含量取决于食品的类型,可以是50mg/kg或更多、更优选100mg/kg或更多、200mg/kg或更多、500mg/kg或更多,最优选200-5000mg/kg。
优选,本发明的食物制品是涂抹料、肉制品、焙烤制品、早餐谷物制品、饮料或条状食品。这些制品是优选的,因为它们的食用,相比其它食物制品,可以达到更受控制地和更恒定地摄取抑制素和大豆蛋白的方式。本发明更优选的食物制品是涂抹料、条状食品或饮料。
下面将通过描述更优选食物制品的适宜的实施方案,来对本发明作进一步的举例说明。使用本文所提供的教导来制备本发明的其它产品是本领域技术人员力所能及的。
肉制品
本发明的肉制品通常具有已知肉制品的组成,然而肉制品中的一部分蛋白质是含抑制素的大豆蛋白。肉制品类型的实例是汉堡包;香肠,如发酵香肠;熟香肠,如法兰克福式香肠;馅饼(pate)等。
发酵香肠的一个实例的组成是:绞碎肉40-70wt%,菌蛋白0-25wt%,5-30wt%含抑制素的发酵大豆蛋白,0.1-1wt%酪蛋白酸钠,0.1-1wt%转谷氨酰胺酶,亚硝酸盐0.5-5wt%,葡萄糖0.1-1wt%,小球菌属培养混合物0.001-0.010wt%和0-1wt%增香剂,盐等等。
法兰克福式香肠的一个实例的组成是:10-40wt%绞碎猪肉,5-20wt%绞碎牛肉,5-25wt%含抑制素的发酵大豆蛋白,0-3wt%盐,0-2wt%亚硝酸盐,0-3wt%磷酸盐,20-50wt%水。
肉制品的典型量值可以是20-200g,通常是40-100g。这种产品中抑制素和蛋白质的优选的含量是:15-350mg/kg抑制素和30-300g/kg大豆蛋白。对这些含量来说,更优选的范围分别是50-200mg/kg抑制素和50-400 g/kg大豆蛋白。
饮料
本发明的优选的食物制品是饮料,例如茶、果汁、软饮料、代餐饮品等。
饮料可以是即饮饮品,或者是需要加入水才能制备成即饮饮料的粉状饮料。
代餐饮品将在下面作更详细描述。显然,对于其它含抑制素和大豆蛋白的饮料来说可以使用相似的含量和组成。一份饮料的典型量值是取200ml。
代餐饮品一般来说是以液体基料为基础,可以例如借助于树胶或纤维将其增稠并且可以向其中添加矿物质和维生素的混合物。可以将这种饮品调味至合意的味道,如果味或巧克力味。其一份的典型量值可以为330ml或330克。
代餐饮料的一个实例的组成是:约70-85wt%水,2-10wt%含抑制素的大豆蛋白,5-15wt%糖(例如蔗糖),0-4wt%脂肪,0.1-1wt%维生素和矿物质,0.01-2wt%风味剂,0.1-0.5wt%增稠剂。
饮料中抑制素和蛋白质的优选含量是:20-100mg/kg抑制素和20-80g/kg大豆蛋白。对这些含量来说,更优选的范围分别是30-70mg/kg抑制素和30-50g/kg大豆蛋白。
条状食品,包括谷物制条状食品
这种产品中经常含有可食物料的基质,其中可以掺加抑制素和大豆蛋白。例如基质可以是脂肪基料的(例如糖皮或巧克力)或者可以是以焙烤制品为基料的(面包,面团,曲奇等)。优选,食物制品是谷物制条状食品,其中基质是以附聚的谷物颗粒为基料(稻米,谷粒,果仁,葡萄干,水果颗粒)。
条状食品的基质物质的量可以占条状食品重量的60-95wt%,优选70-90wt%,最优选75-85wt%。以条状食品的总重量计,基质物料中可以含有5-30wt%的含抑制素的大豆成分,优选10-25wt%,更优选15-25wt%。
谷物制条状食品中的其它成分可以是淀粉、糖(例如0-10wt%)、糖浆、蜂蜜、水、乳固体(0-10wt%)、盐(例如0-5wt%)、碳酸钙(例如0-5wt%)、维生素、增香剂和着色剂。
通常,将成分混合并且熟化(例如,通过熟化挤出),形成需要的形状和尺寸,来生产(谷物制)条状食品。
条状食品的典型量值可以是20-100g,通常是40-60g。条状食品中抑制素和蛋白质的优选含量是:50-500mg/kg抑制素和50-300g/kg大豆蛋白。对这些含量来说,更优选的范围分别是100-350mg/kg抑制素和80-250g/kg大豆蛋白。
乳制品型制品
本发明的乳制品的实例是乳制涂抹料、乳脂干酪、乳型饮品和酸奶,其中乳固体部分或全部地被大豆发酵成分、优选发酵的大豆蛋白分离物所代替。
酸奶型制品的一个实例的组成是:约50-80wt%水,3-12wt%含抑制素的大豆蛋白,0-15wt%乳清粉末,0-15wt%糖(例如,蔗糖),0.01-1wt%酸奶培养物,0-15wt%水果,0.05-0.5wt%维生素和矿物质,0-2wt%风味剂,0-5wt%稳定剂(增稠剂或胶凝剂)。
一份酸奶型制品的典型量值可以是125-250g,通常是150-225g。酸奶型制品中抑制素和大豆蛋白的优选含量是:40-200mg/kg抑制素和20-100g/kg大豆蛋白。对这些含量来说,更优选的范围分别是40-80mg/kg抑制素和30-60g/kg大豆蛋白。
冷冻糖食制品
本发明中,术语“冷冻糖食制品”包括含乳的冷冻糖食,如冰淇淋、冻酸奶、冰糕、加有果酒的冰山(sorbet)、冰乳和蛋奶冻、冰果子露、granitas和冻果泥。
冷冻糖食中的固体的含量(例如糖,脂肪,增香剂等等)优选大于3wt%,更优选从10至70wt%,例如40-70wt%。
冰淇淋中一般含有2-20wt%脂肪、2-20wt%含抑制素的大豆成分、甜味剂、0-10wt%非脂肪乳组分和非必须的组分,诸如乳化剂、稳定剂、防腐剂、增香成分、维生素、矿物质等,其余是水。一般来说,冰淇淋是经过充气的,例如至膨胀度为20-400%,更特别是40-200%,并且冷冻至-2至-200℃,更特别是-10至-30℃。冰淇淋中正常情况下含有约0.1wt%的钙。
本领域技术人员以普通知识为基础,使用适宜量的含抑制素的大豆蛋白作为成分,可以制备其它本发明的食物制品。这些食物制品的实例是焙烤品、乳制品型食品、小吃(例如,烤果仁)等等。
食物制品的制备
根据本发明,将从大豆和/或其成分,如脱脂大豆、大豆蛋白浓缩物或分离物、质构化大豆蛋白等中制备的底物,用丝状真菌发酵,并且使用这种发酵产物来制备食物制品。下面将举例说明这些步骤。在此举例说明中,丝状真菌是红曲霉属真菌。
发酵按已知方式进行。发酵在至少一个发酵容器(发酵罐)中进行,其中在发酵容器中存在含有大豆和/或其成分的培养基。通过添加红曲霉属真菌孢子的悬浮液(接种物)来使发酵开始进行(接种),其中所说的悬浮液是通过将红曲霉真菌在分离(separate)培养基中发酵制成。发酵可以间歇式进行或者以连续式过程来进行。
发酵包括以下步骤,按所给的顺序进行:
a)制备接种物用的培养基和亟待在发酵罐中使用的培养基
b)将培养基、发酵罐和辅助设备灭菌
c)生产接种物
d)在发酵罐中,向含有大豆和/或其成分的培养基中添加接种物
e)进行发酵
f)从发酵罐中取出发酵产物。
将此发酵产物用于制备本发明的食物制品。
非必须地,在将发酵产物用于制备食物制品之前,可以进行以下附加的加工步骤:
g)将发酵产物灭菌
h)将发酵产物(或经过灭菌的发酵产物)干燥
i)进行一步或更多步分离步骤,例如萃取,以便从发酵产物中的红曲霉生物质中分离抑制素和大豆活性物。
发酵罐中所用的培养基可以是固体或液体。有利地,培养基是固体,首选的培养基基本上由经过破碎的整大豆组成(将其用水浸泡直至含水量达20-35wt%水为止)。当培养基是液体时,通常是存在水作为培养基的主要构成成分。
优选使用整大豆作为发酵用的培养基。整大豆的典型组成是42wt%蛋白质、20wt%油、35wt%总碳水化合物、5wt%灰分和5.5wt%粗纤维(Kawamura,S.,Tech.Bull.Faculty Agric.,Kagawa Univ.,18,117(1967))。
可以在发酵罐用的培养基中使用大豆的部分或成分来代替整大豆,例如使用大豆蛋白(包括质构化的植物蛋白质)、大豆乳、大豆薄片等。需要注意的是,培养基中应当含有可以提供利于红曲霉属真菌生长的碳源和氮源的化合物。
本发明中所用的红曲霉属真菌可以是能够产生抑制素的任何红曲霉属真菌。优选真菌选自红色红曲霉。首选的是红色红曲霉F125M1-4。
菌株F125和F125 M1-4保藏在the Centraal Bureau voorSchimmelculturen(CBS),保藏号为CBS 109070(2000年11月14日保藏)和CBS 109269(2001年1月23日保藏)。
这些保藏是布达佩斯条约规定下,根据微生物保藏的国际承认,用于专利程序及其细则目的而作出的(布达佩斯条约)。
保藏材料的利用不能解释为是在违背任何政府的主管部门根据其专利法所授权的权利的前提下实践本发明的许可。
通常情况下,在发酵前要对培养基进行灭菌,例如通过热处理,如巴氏灭菌法。
发酵罐中的培养基可以含有其它可能有助于发酵的物质,例如糖、氨基酸和维生素。
发酵可以按本领域技术人员根据发酵技术普通常规知识来确定的方式进行。下面描述一个优选的实施方案作为举例说明。
发酵温度是重要的。温度优选在10-37℃的范围内,更优选20-30℃。我们发现,在37℃和更高温度下,抑制素的产生降低。
优选,在发酵过程中,给培养基充气,例如通过搅拌、摇动等。充气可以通过让空气吹过发酵培养基来实施。优选当采用固态发酵时,将空气用水蒸气完全或部分地饱和。这样可以避免发酵培养基的干燥。
抑制素对大豆活性物的相对含量取决于发酵的时间。因此发酵时间依发酵产物中抑制素的所需量而定。优选的发酵时间是1-60天,更优选1-50天,进一步优选15-40天,最优选20-30天。
在添加到食物制品中之前,可以对发酵产物进行分离步骤的处理,以便从发酵产物中的红曲霉生物质中分离抑制素和大豆活性物。
这种分离可以用已知的分离技术来进行,例如过滤或离心。
还可以将发酵产物萃取,并且将萃取物用于制备食物制品。
可以对发酵产物进行萃取。或者,可以在萃取之前将红曲霉生物质从发酵产物中分离,例如通过过滤。可以对这种红曲霉生物质单独进行萃取,并且所得的萃取物也可以用于制备食物制品。
可以将萃取物原样直接用于制备食物制品。优选,可以从萃取物中除去萃取溶剂,例如将萃取溶剂蒸发。或者,可以将发酵产物稀释,例如通过添加大豆成分。
有利地,可以使用植物油作为萃取剂。当用植物油,如大豆油,萃取发酵产物时,据发现抑制素可被有效萃取并且可获得含有基本上所有抑制素的油相。所得的萃取物,即植物油,非常适合直接用作食品成分。
最有利的是,在发酵过程中,将萃取剂,例如植物油,添加到发酵培养基中。我们发现,在萃取剂的存在下,发酵过程中抑制素的产生有可观地增加。通过在发酵过程中添加植物油,相比不加萃取剂的发酵,可以使所产生的抑制素的量增加至少10倍,更优选至少40倍。优选萃取剂应当不干扰发酵,特别是它对丝状真菌而言应当不是有毒的。
在本发明的制备食物制品的过程中,可以将这种发酵产物(包括萃取物等)直接添加到食物制品成分中。然而,可以要求将发酵产物用巴氏法灭菌、干燥等,以便将其保存或改良,之后进一步用于制备食物制品。可以将其添加到食物制品组合物的其它成分中,或者可以将它添加到一部分成分中,然后加入其它成分。如果食物制品中存在有一个以上的相,则发酵产物可以存在于这些相的一个或更多个相中。优选,发酵产物基本上存在于油相中,如果这种油相存在的话。
本发明食物制品中的大豆蛋白含量可以按照Agater等,J.Sci.Food Agric.(1986),37(3),317-31中描述的方法测定。
本发明将在以下的实施例中作进一步举例说明。
实施例
实施例1
A.制备红曲霉菌株F125 M1-4
将红色红曲霉菌株F125在麦芽汁水液体培养基中于30℃下培养4天。从此培养物中,取1ml作为用于Hybond-N滤器(Amersham,UK)的接种物使用,将其中所说的滤器放在YE平板上(4%葡萄糖,0.3%KH2PO4,1.0%酵母提取物(Difco),1.5%琼脂)。经过3天的在30℃下的保温培养后,通过用10ml含0.1%吐温80的生理盐水洗涤滤器,收获得孢子。将孢子通过Mira滤布过滤4次,获得不含菌丝的孢子悬浮液。使用此悬浮液用于随后的诱变。
将孢子稀释至108个孢子/ml的浓度,然后暴置于100焦耳/m2强度的UV光下。将此诱变处理的孢子放在马铃薯葡萄糖琼脂(Oxoid)上,并且保温培养3周。选择所得集落中的一个比其它集落颜色较浅的集落,并且在此定义作红曲霉菌株F125 M1-4。
将该菌株F125 M1-4于2000年11月14日保藏在the CentraalBureau voor Schimmelculturen(CBS)并且保藏号为CBS 109070。
B.制备红曲霉孢子
通过从PDA(马铃薯葡萄糖琼脂)(oxoid)斜面培养基中收获红曲霉菌丝体,来制备红曲霉f125M1-4孢子,即用5ml生理盐水洗涤斜面培养基并且将菌丝体在150ml麦芽汁水(oxoid)中于30℃下保温培养4天。通过Mira滤布过滤收获得到孢子。
C.发酵
C.1.制备接种物
在含有质构化植物蛋白质(TVP)的摇瓶中,接种悬浮于含0.1wt%吐温80(聚氧乙烯脱水山梨醇脂肪酸酯,得自ICI SpecialtyChemicalsTM)生理盐水溶液中的丝状孢子。将此摇瓶保温培养,让霉菌生长。由此得到红曲霉孢子悬浮液,通过稀释调整至1×106个孢子/ml的浓度。
C.2.制备发酵培养基
将1kg TVP浸泡在自来水(50℃)中30分钟。浸泡过后,将TVP小碎块用冷自来水漂洗。随后,将50g批量的浸泡TVP(35wt%干物质)送入300ml锥形烧瓶中。
C.3.发酵
给每个摇瓶接种1ml的制备的红曲霉孢子悬浮液并且在30℃下保温培养30天。每周从摇瓶中取样,来监测抑制素的产生。
发酵后,剩留约600g质构化大豆蛋白(含水量28%)。将此大豆发酵成分在85℃下巴氏灭菌30分钟,冷却并且储存用于制备食物制品。
D.发酵产物的抑制素分析
将发酵产物样品在50ml管(Falcon)中,通过添加6ml乙腈、水和磷酸的混合物(1∶1∶0.05,v/v/v)来萃取。将混合物与Ultraturrax共混1分钟。然后,将混合物在室温下于rollerbank上保温培养24小时以上。之后将样品离心并且使用上清液用于HPLC分析。在Shimadzu仪器上按照Morovian等,J.chromatogr.A 763(1997)165-172中所述的方法,使用HPLC分析来分离样品。该系统由Shimadzu SCL-10A系统控制器、CTO-10AS柱烘箱、LC-10AT vp泵系统、RID10A折射率检测器、SPD-M1OA二极管阵列检测器和SIL-10AD自动进样器组成。对于色谱测定抑制素,使用Waters NovaPak C18(150×3.9mm I.D.,4μm)柱,在25℃下操作。洗脱液是乙腈-0.1%磷酸(50∶50,v/v)溶液,流速为1.5ml/min。洗脱进行15分钟。使用二极管阵列检测器从190nm至最高800nm进行检测。测定光谱中属于抑制素的所有峰的面积的总和。与标准样(Mevinolin,Sigma)比较,计算抑制素含量(按mg/kg被分析产物计)。
分析结果得到抑制素含量为1200mg/kg。
F.制备发酵香肠
制备具有如下组成的发酵香肠:
表4:发酵香肠的组成
成分 | wt%样品a | wt%样品b |
红曲霉发酵的TVP | 10 | 20 |
Quorn(菌蛋白基料的肉代用品,Marlow foods Limited制造) | 25 | 15 |
盐腌的绞碎猪肉 | 35.6 | 35.6 |
盐腌的腌肉 | 25 | 25 |
酪蛋白酸Na | 0.4 | 0.4 |
转谷氨酰胺酶活性物 | 0.4 | 0.4 |
亚硝酸盐 | 1.96 | 1.96 |
抗坏血酸钠 | 0.06 | 0.06 |
葡萄糖 | 0.4 | 0.4 |
小球菌属发酵剂,Quest Int.出品 | 0.016 | 0.016 |
水 | 0.024 | 0.024 |
总计 | 100 | 100 |
发酵香肠的制备是通过将所有成分混合。将所得的混合物灌装至肠衣中并且将香肠在2℃下储存24小时。
然后,在28℃下进行香肠的发酵约24小时,直至pH为5.2为止。将所得的香肠在20℃下干燥4天。香肠的pH为4.7并且干物质含量为72wt%。
通过上述方法分析,此发酵香肠中含有分别为65g/kg(样品a)、135g/kg(样品b)的大豆蛋白和分别为110mg/kg(样品a)、245mg/kg(样品b)的抑制素。
实施例2
按照实施例1进行步骤A、B和D。
C.发酵
C.1.制备接种物
在含有脱皮大豆的摇瓶中,接种悬浮于含0.1wt%吐温80(聚氧乙烯脱水山梨醇脂肪酸酯,得自ICI Specialty ChemicalsTM)生理盐水溶液中的丝状孢子。将此摇瓶保温培养,让霉菌生长。由此得到红曲霉孢子悬浮液,通过稀释调整至1×106个孢子/ml的浓度。
C.2.制备发酵培养基
将1kg脱皮大豆浸泡在自来水(50℃)中30分钟,并且随后用空气干燥2小时。随后,将50g批量的干燥脱皮大豆送入300ml锥形烧瓶中。
C.3.发酵
给每个摇瓶接种1ml制备的红曲霉孢子悬浮液并且在30℃下保温培养30天。每周从摇瓶中取样,来监测抑制素的产生。
发酵后,剩留约600g大豆。根据步骤D的分析,抑制素含量为2800mg/kg产物。
E.发酵产物的异黄酮分析
按照Franke A.A.,等(1998):食品和人流体中异黄酮类化合物和其它酚类试剂的HPLC分析(HPLC analysis of isoflavonoids andother phenolic agents from foods and human fluids);Proceed.Soc.Exp.Biol.Med;217(3),274-280中描述的方法测定异黄酮浓度。
测试两种样品。第一种(对比)样品取自如上步骤C2所制备的未发酵的发酵培养基。第二种样品为相等(固体)重量,但取自发酵产物。这些样品的异黄酮测定结果示于下表3。
表3.发酵和未发酵大豆中异黄酮的浓度
异黄酮 | 异黄酮浓度(g/kg) | |
发酵培养基(未发酵的) | 发酵产物 | |
大豆苷 | 1.107 | 0.304 |
染料木苷 | 1.608 | 0.476 |
大豆黄酮 | 0.057 | 0.9 |
染料木素 | 0.085 | 0.494 |
总计 | 2.9 | 2.2 |
表3显示了发酵产物中的总的异黄酮的量比未发酵物料有所降低,但是意想不到的是,染料木素和大豆黄酮的量有所增加。发酵产物中除抑制素外还含有显著量的异黄酮。发酵产物中的抑制素含量为2800mg/kg、蛋白质含量为27.8wt%,它含有900mg/kg大豆黄酮和494mg/kg染料木素,适合作为食品成分。
Claims (10)
1.每平均一份中含有至少5克大豆蛋白的食物制品,其特征在于,该食物制品中含有至少5mg/kg的抑制素。
2.权利要求1的食物制品,其特征在于,食物制品中含有大豆发酵成分。
3.含有抑制素、大豆蛋白以及染料木素和染料木苷的食物制品,其中,抑制素的量为5-500mg/kg,大豆蛋白的量为10-500g/kg,并且染料木素的量为染料木素和染料木苷总量的10-99wt%。
4.权利要求3的食物制品,其中,染料木素的量为染料木素和染料木苷总量的20-80wt%。
5.权利要求1-4任一项的食物制品,其中,大豆发酵成分是一种或多种丝状真菌的发酵产物,其中所说的丝状真菌选自红曲霉属、曲霉属、青霉属、灰侧耳菌属、腐霉属、隐壳霉属、拟青霉属、Eupenicillium和束孢霉属。
6.权利要求5的食物制品,其特征在于,大豆发酵成分是红曲霉的发酵产物。
7.权利要求1-6任一项的食物制品,其中,食物制品是涂抹料、肉制品、焙烤制品、饮料或条状食品。
8.权利要求7的食物制品,其中,食物制品是条状食品、饮料或肉制品。
9.权利要求8的食物制品,其中,食物制品是具有如下组成的肉制品:绞碎肉60-80wt%,菌蛋白15-25wt%,2-10wt%含抑制素的大豆蛋白,0.1-1wt%酪蛋白酸钠,0.1-1wt%转谷氨酰胺酶,亚硝酸盐0.5-5wt%,葡萄糖0.1-1wt%,小球菌属培养混合物0.001-0.010wt%0-1wt%和非必须的其它成分。
10.权利要求8的食物制品,其中,食物制品是具有如下组成的肉制品:10-40wt%绞碎猪肉,5-20wt%绞碎牛肉,2-10wt%含抑制素的大豆蛋白,0-5wt%盐,0-5wt%亚硝酸盐,0-5wt%磷酸盐,20-50%水和非必须的其它成分。
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WO2005058054A2 (en) * | 2003-12-18 | 2005-06-30 | Unilever Plc | Confectionery product |
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BRPI0513939A (pt) * | 2004-08-23 | 2008-05-20 | Unilever Nv | composição, formador de creme e/ou clareador, composição de alimento, processo para a manufatura de uma composição e uso de uma composição |
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