CN103327827B - 用于活性成分的保护性水胶体 - Google Patents
用于活性成分的保护性水胶体 Download PDFInfo
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- CN103327827B CN103327827B CN201180066052.3A CN201180066052A CN103327827B CN 103327827 B CN103327827 B CN 103327827B CN 201180066052 A CN201180066052 A CN 201180066052A CN 103327827 B CN103327827 B CN 103327827B
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- protein
- sorghum
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Abstract
(经修饰的)高粱蛋白质被用作用于活性成分的新颖保护性水胶体,所述活性成分尤其是脂溶性活性成分和/或着色剂。本发明包括组合物及其生产方法,所述组合物包含(经修饰的)高粱蛋白质和至少一种活性成分,本发明还包括(经修饰的)高粱蛋白质本身及其生产方法。这些组合物用于食品、饮料、动物饲料、个人护理组合物和药物组合物的富集、强化和/或着色,本发明还涉及分别包含这种(经修饰的)高粱蛋白质和这种组合物的食品、饮料、动物饲料、个人护理组合物和药物组合物。
Description
本发明涉及(经修饰的)高粱蛋白质作为活性成分的新颖保护性水胶体的用途,所述活性成分尤其是脂溶性活性成分和/或着色剂。此外,本发明涉及下述组合物及其生产方法,所述组合物包含(经修饰的)高粱蛋白质和至少一种活性成分,本发明还涉及(经修饰的)高粱蛋白质本身及其生产方法。本发明还涉及此类组合物用于对食品、饮料、动物饲料、个人护理组合物或药物组合物进行富集、强化和/或着色的用途,以及分别含有此类(经修饰的)高粱蛋白质和此类组合物的食品、饮料、动物饲料、个人护理组合物和药物组合物。
活性成分,尤其是脂溶性活性成分或着色剂,通常不原样添加至食品、饮料、动物饲料、个人护理组合物和药物组合物中,而是以保护性水胶体中活性成分制剂的形式添加,这是为了增强例如化学稳定性、(水)溶解度、自由流动性和受控释放等性质。已知的保护性水胶体为例如不同来源(禽、牛、猪、鱼)的明胶和淀粉。由于宗教方面或过敏反应方面的原因,动物来源的保护性水胶体通常是人们不期望的,而对无麦麸(gluten)和无玉米(corn)产品感兴趣的消费者不喜欢基于淀粉的保护性水胶体,因此,人们仍需要替代性的保护性水胶体。尽管如此,保护性水胶体应该具有良好的营养值和广泛的应用领域。
现已发现高粱蛋白质可以被用作这种保护性水胶体。
在许多国家,高粱是一种重要的粮食作物。有许多种高粱,例如黑高粱(sorghumalmum)、sorghumamplum、sorghumangustum、sor-ghumarundinaceum、二色高粱(sorghumbicolour)、sorghumbicolorsubspeciesdrummondii(苏丹草)、sorghumbrachypodum、sorghumbulbosum、sorghumburmahicum、sorghumecarinatum、sorghumexstans、sorghumgrande、假高粱(sorghumhalepense)(詹森草)、sorghuminterjectum、sorghumintrans、sorghumlaxiflorum、sorghumleiocladum、sorghummacrospermum、sorghummatarankense、光高粱(sor-ghumnitidum)、sorghumplumosum、拟高粱(sorghumpropinquum)、sorghumpurpureosericeum、sorghumstipoideum、sorghumtimorense、sorghumtrichocladumand多色高粱(sorghumversicolor)。其中最常见的高粱之一是二色高粱。
二色高粱是一种重要的世界作物,用于食品(作为谷物,以及用于高粱糖浆或“高粱糖蜜”中)、草料、酒精饮料的生产、以及生物燃料。大多数品种是耐旱耐热的,并且在谷物是穷人或农村居民的主食或主食之一的干旱地区是特别重要的。在许多热带地区,它们形成了牧场的重要组成部分。高粱在非洲、中美洲和南亚是重要的粮食作物,并且是全世界第五重要的谷类作物。
高粱的蛋白质含量可以改变。但通常在9重量-%和15重量-%之间。
高粱蛋白质在营养品质方面排名很高。高粱蛋白质被认作是有营养的且低过敏性的,并因此可以是用于活性成分制剂的保护性水胶体的合适的替代性来源。然而,在中性pH下高粱蛋白质的高度不溶性和较差的功能性限制了其作为功能性成分在食品和药物产品中的工业应用。本发明克服了这些限制,并包含(经修饰)的高粱蛋白质作为活性成分(尤其是脂溶性活性成分和/或着色剂)制剂的保护性水胶体。
占主要地位的高粱蛋白质是疏水的。可以从高粱籽粒的核心(胚乳蛋白)以及从其他部分分离蛋白质。
提取的蛋白质本质上是高度不可溶的,并且在蛋白质分离中所使用的条件进一步降低它们的溶解度,由此它们在作为功能性成分方面的应用是受限制的。可通过碱提取然后在蛋白质的等电点pH下沉淀,来从高粱粉中获得高蛋白的高粱产品。通常使用淀粉水解酶,例如α-淀粉酶、葡糖淀粉酶和支链淀粉酶(pullulanase),通过溶解和除去淀粉来分离高粱粉中的蛋白质。除了淀粉水解酶之外,还已经用纤维素酶和半纤维素酶来进一步增加高粱蛋白质浓缩物中的蛋白质含量。然而,关于合适的提取方法和此类分离物功能性的信息是有限的。使用经批准的食品级酶和化学物质的高效提取方法对于高粱蛋白质的商业生产和应用来说是必要的。
包含高粱蛋白质和活性成分的本发明的组合物满足了这种需要。
发明详述
本发明的组合物可以是固体组合物,即稳定的水溶性粉末或水可分散性粉末,或者它们可以是液体组合物,即上述粉末的水性胶体溶液或水包油分散体。可通过下文所述的方法或通过类似手段,来制备经稳定的水包油分散体,其可以是水包油乳液或者可以是悬浮的(即,固体)颗粒和乳化的(即,液体)液滴的混合物。
更具体地,本发明涉及粉末形式的稳定组合物,其包含处于(经修饰的)高粱蛋白质基质中的一种或多种(脂溶性)活性成分和/或一种或多种着色剂。
优选地,基于组合物的总量,(经修饰的)高粱蛋白质的量为1至70重量%、更优选地5至50重量%、甚至更优选地10至40重量%、最优选地10至20重量%(其中20重量%是最优选的量),和/或,(脂溶性)活性成分和/或着色剂的量为0.1至90重量%、优选地1至80重量%、更优选地1至20重量%。如果存在额外佐剂和/或赋形剂,例如生育酚和/或棕榈酸抗坏血酸酯,它们以基于组合物总量的0.01至50重量%的量、优选地以0.1至30重量%的量、更优选地以0.5至10重量%的量存在。
(经修饰的)高粱蛋白质
在本发明的一些优选实施方式中,高粱蛋白质是经修饰的高粱蛋白质,其生产方法如下文所述。一种尤其优选的高粱蛋白质是通过下述步骤获得的:碱提取,(酶促修饰,尤其是使用碱性蛋白酶的酶促修饰),离心和超滤。如果进一步用途需要,还可以干燥由此获得的经修饰的高粱蛋白质。
然而,本发明并不仅限于由此生产的经修饰的高粱蛋白质的用途。
甚至更优选的是具有≥220ml/g、优选≥350ml/g、更优选≥500ml/g、甚至更优选500ml/g至1000ml/g的乳化能力的(经修饰的)高粱蛋白质。此外,在本发明的优选的实施方式中,所用的(经修饰的)高粱蛋白质具有≥0.2、优选地≥0.45、更优选地≥0.5、甚至更优选地0.5至1.0的乳化活性。对乳化能力的测定以及对乳化活性的测定如下所述。本发明还涉及这些(经修饰的)高粱蛋白质本身。
乳化能力的测定
按照Gbogouri等JournalofFoodScience2004第69卷,Nr.8,615的方法,基于油滴定,来测定(经修饰的)高粱蛋白质的乳化能力。在去离子水中制备(经修饰的)高粱蛋白质的分散体(0.1%w/w,50mL,pH7.0)。用设定为1的均化器(VirtishearTempest,TheVirtisCo.,Gardiner,N.Y.,U.S.A.)来均化蛋白质溶液。使用蠕动泵,以大约17g/min的流速向蛋白质溶液中加入玉米油。通过电导率仪连续记录乳液的电导率,将其用作测定乳化转化点(inversionpoint)的参数。用加至转化点的油的量来计算乳化能力。乳化能力被表示为乳化的油减去空白后相对于样品中蛋白质的量的比。空白是在50mL去离子水中相转化之前加入的油的量。
或者可以按照Vuillemard和他人的方法,基于油滴定测定乳化能力。在蒸馏水中制备蛋白质分散体(0.5%w/w,40mL,pH7.0)。用设定为6的均化器(VirtishearTempest,TheVirtisCo.,Gardiner,N.Y.,U.S.A.)来均化蛋白质溶液。使用泵,以12g/min的流速向蛋白质溶液中加入玉米油。通过电导率仪连续记录乳液的电导率,将其用作测定乳化转化点的参数。用加至转化点的油的量来计算乳化能力。乳化能力被表示为乳化的油减去空白后相对于样品中蛋白质的量的比。空白是在40mL蒸馏水中相转化之前加入的油的量。
乳化活性的测定
通过PearceandKinsella,JournalofAgricFoodChem.1978,26:716-722的浊度方法来测定乳化活性。用设定为6的超声仪(VirtishearTempest,TheVirtisCo.,Gardiner,N.Y.,U.S.A.),对6ml0.1%的(经修饰的)高粱蛋白质在10mMpH7.0的磷酸盐缓冲液中的溶液与2ml玉米油的混合物进行1分钟的均化。均化后0和10分钟时,将50微升混合物转入5ml0.1%(w/v)的SDS水溶液中。用分光光度计(ShimadzuModelUV-1601,Kyoto,日本)测定500nm处溶液的吸光度。均化后时间0时的吸光度就是(经修饰的)高粱蛋白质的乳化能力。
在本发明的其他优选的组合物中,(经修饰的)高粱蛋白质与至少一种选自由还原糖、糖蛋白和糖肽组成的组的化合物交联。
活性成分
一般而言,活性成分是具有药理学作用的那些成分或者对人或动物体提供健康益处的那些。优选地,活性成分是脂溶性活性成分和/或着色剂。
脂溶性活性成分和/或着色剂优选地选自由以下组成的组:胡萝卜素和结构相关的多烯化合物、脂溶性维生素、辅酶Q10、多不饱和脂肪酸(例如二十碳五烯酸(EPA)和二十二碳六烯酸(DHA))及其酯(例如,乙酯或甘油三酸酯(含有相同或不同的脂肪酸))、富含多不饱和脂肪酸的甘油单/二/三酸酯、脂溶性UV-A滤光剂、UV-B滤光剂以及其生理学可接受的衍生物(例如其酯,尤其是与C1-20碳酸的酯)以及它们的任何混合物。
最优选的脂溶性维生素是维生素A或维生素E(以及它们的衍生物)。
胡萝卜素和结构相关的多烯化合物的优选的例子是类胡萝卜素,例如α-胡萝卜素、β-胡萝卜素、8’-阿朴-β-胡萝卜醛(carotenal)、8’-阿朴-β-胡萝卜素酸酯(例如乙酯)、斑蝥黄(canthaxanthin)、虾青素、番茄红素、叶黄素、玉米黄质、藏红花酸、α-玉米胡萝卜素(zeacarotene)、β-玉米胡萝卜素及它们的生理学可接受的衍生物(例如它们的酯,尤其是与C1-20碳酸的酯)以及它们的任何混合物。
最优选的类胡萝卜素是β-胡萝卜素。
术语“β-胡萝卜素”包括全顺式异构体和全反式异构体以及所有可能的混合顺反异构体。这同样应用于其它类胡萝卜素。
术语“玉米黄质”包括天然的R,R-玉米黄质以及S,S-玉米黄质、内消旋玉米黄质以及它们的任何混合物。这同样适用于叶黄素。
(脂溶性)活性成分可以是天然来源的,即,从植物中分离/提取的、纯化和/或浓缩的,以及通过化学和/或微生物(发酵)途径合成的那些。
其它组分
除了活性成分和(经修饰的)高粱蛋白质之外,优选地,本发明的组合物还可含有至少一种水溶性抗氧化剂和/或脂溶性抗氧化剂。
水溶性抗氧化剂可以例如是抗坏血酸或其盐(优选抗坏血酸钠),水溶性多酚,例如羟基酪醇(hydroxytyrosol)和橄榄多酚配醣体(oleuropeinaglycon);表没食子儿茶素没食子酸酯(EGCG)或橄榄或迷迭香的提取物。
脂溶性抗氧化剂可以例如是生育酚,例如dl-α-生育酚(即,合成生育酚)、d-α-生育酚(即,天然生育酚)、β-生育酚或γ-生育酚或它们中两种或多种的混合物;丁基化羟基甲苯(BHT);丁基化羟基茴香醚(BHA);乙氧喹啉、没食子酸丙酯;叔丁基羟基喹啉;或6-乙氧基-1,2-二羟基-2,2,4-三甲基喹啉(EMQ)或脂肪酸的抗坏血酸酯,优选棕榈酸抗坏血酸酯或硬脂酸抗坏血酸酯。
本发明的组合物还可包含选自下组的共乳化剂(co-emulgator),所述组由以下组成:脂肪酸的甘油单酸酯和二酸酯、脂肪酸的多甘油酯、卵磷脂;N-酰化氨基酸及其衍生物、具有烷基或烯基的N-酰化肽及其盐;烷基或烯基醚或硫酸酯及其衍生物和盐;聚氧乙烯化烷基或烯基脂肪醚或酯;聚氧乙烯化烷基或烯基羧酸及其盐;N-烷基或N-烯基甜菜碱;烷基三甲基铵或烯基三甲基铵及其盐;多醇烷基或烯基醚或酯;及其混合物。
多醇烷基或烯基醚或酯的优选的例子是被至少20个环氧乙烷单元聚氧乙烯化的山梨聚糖烷基或烯基酯,例如,山梨聚糖棕榈酸酯20EO或聚山梨糖40(Seppic公司以商品名Montanox40DF销售)、山梨聚糖月桂酸酯20EO或聚山梨糖20(ICI公司以商品名Tween20销售)和山梨醇酐单硬脂酸酯。
根据本发明的制剂还可被压成片剂,其中可以加入选自由单糖、二糖、寡糖和多糖、甘油和甘油三酸酯组成的组的一种或多种赋形剂和/或佐剂。
可存在于本发明组合物中的单糖和二糖的优选的例子是蔗糖、转化糖、木糖、葡萄糖、果糖、乳糖、麦芽糖、甘蔗糖(saccharose)和糖醇。
寡糖和多糖的优选的例子是淀粉、改性淀粉和淀粉水解产物。淀粉水解产物的优选的例子是糊精和麦芽糊精(尤其是具有5至65范围内的葡糖当量(DE)的那些)和葡萄糖浆(尤其是具有20至95范围内的DE的那些)。术语“葡糖当量”(DE)指水解程度,其是还原糖的量的量度,被计算为基于干重的D-葡萄糖;该标度基于:DE接近于0的天然淀粉和DE为100的葡萄糖。
合适地,甘油三酸酯是植物油或脂肪,优选玉米油、葵花籽油、大豆油、红花油、油菜籽油、花生油、棕榈油、棕榈仁油、棉籽油、橄榄油或椰油。
固体组合物还可以包含抗结块剂(例如硅酸或磷酸三钙等)以及至多10重量%、通常为2-5重量%的水。
组合物的制造
本发明的一个目的还是制造本发明组合物的方法,所述方法包括下述步骤:
I)制备(经修饰的)高粱蛋白质的水性溶液或胶体溶液,
II)任选地,向步骤I)中制备的溶液中至少加入水溶性赋形剂和/或佐剂,
III)制备至少有活性成分(优选地,至少有脂溶性活性成分和/或着色剂)以及任选地至少脂溶性佐剂和/或赋形剂的溶液或分散体,
IV)将步骤I)至III)中制备的溶液互相混合,
V)将由此得到的混合物均化,
VI)任选地加入用来交联(经修饰的)高粱蛋白质的交联剂,
VIa)任选地在已经进行步骤VI)后将所获得的混合物进行酶促处理或热处理,来交联(经修饰的)高粱蛋白质,
VII)任选地将步骤V)和/或VI)中获得的分散体转化成粉末,
VIII)任选地干燥步骤VII)中所获得的粉末,
IX)任选地对干燥的粉末进行热处理或酶促处理,来交联(经修饰的)高粱蛋白质,
同时满足下述条件:在进行步骤VI)的情况下,仅进行步骤VIa)或步骤IX)之一,而非两者都进行。
作为替代性方法,还可以在修饰后但乳化步骤前交联蛋白质。
步骤I
该步骤通过将水加入(经修饰的)高粱蛋白质或者反过来操作来简单地进行,任选地,在搅拌下进行。或者,可以通过超声处理来进行均化。
优选地,使用具有上文所述的优选级(preferences)的(经修饰的)高粱蛋白质。
步骤II
可加入水溶性赋形剂和/或佐剂,例如单糖、二糖、寡糖和多糖、甘油和水溶性抗氧化剂。它们的例子在上文中给出。
步骤III
活性成分是上文描述过的那些。
(脂溶性)活性成分和/或着色剂以及任选的脂溶性赋形剂和佐剂或者被原样使用,或者溶解或悬浮于甘油三酸酯和/或(有机)溶剂中使用。
合适的有机溶剂是卤代脂肪族碳氢化合物、脂肪族醚、脂肪族碳酸酯和环状碳酸酯、脂肪族酯和环状酯(内酯)、脂肪族酮和环状酮、脂肪族醇及其混合物。
卤代脂肪族碳氢化合物的例子是单卤代或多卤代的、线性或支链的或环状的C1至C15的烷烃。尤其优选的例子是单氯代或多氯代或单溴代或多溴代的、线性或支链的或环状的C1至C15的烷烃。更优选的是单氯代或多氯代的、线性或支链的或环状的C1至C15的烷烃。最优选的是二氯甲烷和氯仿。
脂肪族酯和环状酯(内酯)的例子是乙酸乙酯、乙酸异丙酯和乙酸正丁酯以及γ-丁内酯。
脂肪族酮和环状酮的例子是丙酮、二乙基甲酮和异丁基甲基甲酮;以及环戊酮和异佛乐酮。
环状碳酸酯的例子尤其是碳酸亚乙酯和碳酸亚丙酯及其混合物。
脂肪族醚的例子是二烷基醚,其中烷基部分具有1至4个碳原子。一个优选的例子是二甲基醚。
脂肪族醇的例子是乙醇、异丙醇、丙醇和丁醇。
此外,任何油(甘油三酸酯)、橙油、柠檬油精(limonen)等和水可被用作溶剂。
可加入的脂溶性赋形剂和/或佐剂是,例如,玉米油、脂肪酸的甘油单酸酯或脂肪酸的甘油二酸酯、多甘油脂肪酸酯以及中链甘油三酸酯(“MCT”)。
步骤IV
在本发明的一种替代性方法中,不进行步骤III),而是直接向步骤I)或II)的溶液中加入活性成分和可选的脂溶性赋形剂和/或佐剂。
步骤V
为进行均化,可以应用常规技术,例如高压均化、高剪切力乳化(转子-定子系统)、微粉化、湿碾磨、微通道乳化、膜乳化或超声波处理。用来制备包含(脂溶性)活性成分和/或着色剂的,用于食品、饮料、动物饲料、化妆品或药物组合物的富集、强化和/或着色的组合物的其它技术公开于EP-A0937412(尤其是[0008]、[0014]、[0015]、[0022]至[0028]段)、EP-A1008380(尤其是[0005]、[0007]、[0008]、[0012]、[0022]、[0023]至[0039]段)和US6,093,348(尤其是第2栏第24行至第3栏第32行;第3栏第48-65行;第4栏第53行至第6栏第60行)中,这些文献的内容都通过引用并入本文。
步骤VI
交联剂优选地选自由还原糖、糖蛋白和糖肽组成的组。因此,(经修饰的)高粱蛋白质和糖蛋白/糖肽的糖或糖部分之间形成分子间交联。交联剂的优选的例子是还原糖,例如葡萄糖、果糖、甘蔗糖和木糖。
步骤VIa
交联可以通过对额外包含上文所述交联剂的混合物进行热处理,使得糖与蛋白质在Maillard型的反应中交联来实现,即,通过热处理,优选在大约30℃至大约160℃的温度、更优选地大约70℃至大约100℃的温度、最优选地大约80℃至大约90℃的温度下进行。
(经修饰的)高粱蛋白质与交联剂的交联还可通过用交联酶(酰基转移酶,EC2.3,例如,转谷氨酰胺酶,EC2.3.2.13,蛋白质-谷氨酰胺:γ-谷氨酰转移酶)处理来实现,即通过酶促处理来实现,其方便地进行于大约0℃至大约70℃的温度下,优选地,大约20℃至大约40℃的温度下。优选地,根据步骤VIa)的酶促处理是用交联酶、特别是转谷氨酰胺酶进行的处理。
酶促交联导致产生稳定的含蛋白质多糖网络,在用转谷氨酰胺酶的情况下,这是通过ε-(γ-谷氨酰)-赖氨酸异肽键的形成来实现的。对于酶促交联来说,使用糖蛋白或糖肽是优选的。
使糖和蛋白质在Maillard型反应中交联的热处理和酶促交联这两种技术都可用于将亲脂性部分并入,并且可以在干燥形式的组合物(步骤IX)或水性溶液或悬浮液(步骤VIa)中进行。酶促交联优选在水性溶液或悬浮液中进行。
步骤VII
由此获得的分散体(其是水包油分散体)可在除去有机溶剂(如果存在的话)之后被转化为固体组合物,例如干粉,这通过使用任何常规技术来进行,例如,喷雾干燥,喷雾干燥与流床制粒(后者通常称为流式喷雾干燥或FSD)的组合或者通过粉末捕捉技术(喷雾的乳液液滴由此被捕捉于吸收剂的床中并随后被干燥,所述吸收剂例如为淀粉、硅酸钙和二氧化硅)来实现。
喷雾干燥可在大约100℃至大约250℃、优选大约150℃至大约200℃、更优选大约160℃至大约190℃的进口温度和/或大约45℃至大约160℃、优选大约55℃至大约110℃、更优选大约65℃至大约95℃的出口温度(产物温度)下进行。
步骤VIII
对步骤VII中获得的粉末的干燥优选在≤100℃的温度下、优选20℃至100℃的温度下、更优选60℃至70℃的温度下进行。如果干燥在真空下进行,温度会更低。
步骤IX
如上文中已经针对步骤VIa所描述的,通过热处理进行交联。这同样应用于酶促处理,但是酶促处理优选在溶液/悬浮液中进行。
(经修饰的)高粱蛋白质的制造
本发明还涉及从经碾磨的高粱开始来制造(经修饰的)高粱蛋白质的方法,所述方法包括下述步骤a)至e),并且满足下述条件:步骤b)、c)和d)中至少要进行一个。可以在碾磨之前除去高粱麸。
所述方法包括:
a)制备经碾磨高粱的水性溶液或悬浮液(其中可在碾磨之前除去高粱麸),其中基于水性溶液或悬浮液的总量,溶液或悬浮液优选地具有0.1至30重量%、优选10至15重量%的干物质含量;
b)任选地移出经碾磨高粱的非蛋白质部分或蛋白质部分(从而在碾磨之前除去高粱麸),来获得高粱蛋白质;
c)任选地修饰经碾磨的高粱的蛋白质部分(其中在碾磨之前可除去高粱麸),来获得经修饰的高粱蛋白质;
d)任选地分离(经修饰的)高粱蛋白质;
e)任选地将(经修饰的)高粱蛋白质转化为固体形式。
在本发明的上下文中,“高粱蛋白质”尤其表示通过进行以下任何所获得的产物:步骤a)和b);或步骤a)和d);或步骤a)、b)和e);或步骤a)、b)和d);或步骤a)、d)和e);或步骤a)、b)、d)和e)。优选地的是进行步骤a)和b)(和d)和/或e))的实施方式,尤其优选的是进行步骤a)、b)和d)(和e))的实施方式。
在本发明的上下文中,“经修饰的高粱蛋白质”尤其表示进行步骤c)获得的产物,即,通过进行以下任何所获得的产物:步骤a)和c);或步骤a)、b)和c);或步骤a)、c)和d);或步骤a)、c)和e);或步骤a)、b)、c)和d);或步骤a)、c)、d)和e);或步骤a)、b)、c)和e);或步骤a)、b)、c)、d)和e)。
步骤a)
经碾磨的高粱(其中在碾磨之前除去高粱麸)也被表示成“高粱粉”。
该步骤通过向高粱粉中加入水或者反过来操作简单地进行,任选地,通过剧烈搅拌(用机械搅拌器)来进行,直到高粱粉完全分散,或者通过用均化器对高粱粉悬浮液进行均化(例如,室温下5分钟)来进行。
步骤b)
移除非蛋白质部分
优选地,步骤b)可这样来实现:用非蛋白质降解酶处理高粱粉,例如,用的0.5%水性悬浮液在90℃下处理2小时,然后用纤维素酶的0.1%水性悬浮液在50℃下处理30分钟——没有任何pH调节(pH6-7),使酶失活,从高粱粉的蛋白质部分分离并移除非蛋白质部分。
非蛋白质降解酶的优选的例子是淀粉降解酶,例如,α-淀粉酶和纤维素酶(即,纤维素降解酶)及其混合物。α-淀粉酶的优选的例子是120,L型,其可从NovoNordiskBiochem,NorthAmerica,Inc.,USA商业获得。其它优选的例子是Supra(可从NovoNordiskBiochem,NorthAmerica,Inc.,USA商业获得)、AmylaseS″Amano″35G(可从AmanoPharmaceuticalCo.Ltd.,Nagoya,Japan商业获得)、MultifectCellulase(可从GenencorInternational,Inc.,USA商业获得)和CellulaseT″Amano″4(可从AmanoPharmaceuticalCo.Ltd.,Nagoya,Japan商业获得)。
如果使用无机酸(例如氢氯酸)或有机酸(例如柠檬酸)或碱时,可通过对溶液或悬浮液加以中和来终止酶反应,或者通过加热使酶变性来终止酶反应。
可通过将溶液加热至80℃至95℃的温度、优选地80℃至85℃的温度(尤其是在3.5至4.5的低pH下)10-15分钟来实现变性。之后溶液可被冷却至50℃。
可通过离心(5000g15分钟)(由此使得非蛋白质部分在水相中),接着用去离子水洗涤来实现对非蛋白质部分的分离。高粱蛋白质留在沉淀团中。
移除蛋白质部分
或者,在离心或过滤之前可以进行所谓的“碱提”或所谓的“盐提”。
“碱提”指的是先在40至60℃用碱溶液(例如NaOH水溶液)将高粱粉溶液或悬浮液的pH调节3小时至7至12的值、优选地8至10的值、更优选地大约9的值。
当蛋白质产率比蛋白质功能性更重要的情况下,将pH优选地调节至8至12、更优选9至12、甚至更优选10至12的数值可能是有利的。
优选地,此类碱具有大约0.1至5M、优选地大约0.5至大约2M的浓度。碱可以是无机碱。无机碱的例子是碱(土)金属氢氧化物,例如氢氧化钠(优选的)、氢氧化钾和氢氧化钙。
“盐提”与“碱提”类似,但是除碱之外,还使用盐,例如氯化钠。在本发明的一种优选实施方式中,使用0.08M氯化钠水溶液(用NaOH调节至pH11)作为提取溶剂。
在两种情况(碱提或盐提)下,蛋白质部分都被转移至水相。然后可通过离心或过滤将蛋白质部分与非蛋白质部分分离。
步骤c)
可通过用(商业可获得的)食品级碱性、中性和/或酸性蛋白酶处理高粱粉(的蛋白质部分),来实现对高粱粉的修饰。实施例中给出了针对一些蛋白酶的酶规格和最优条件。
蛋白酶可以来自细菌或真菌,以及来自水果,或者可以是动物来源的。
碱性蛋白酶的例子是商业可获得的(NovoNordiskBiochem,Franklinton,NC,USA)、Alkaline(EnzymeDevelopmentCorporation,NewYork,NY,USA)、Protex(BacterialAlkalineProtease,GenencorInternational,Inc.,Rochester,NY,USA)和Protease899(GenencorInternational,Inc.,Rochester,NY,USA)。
中性蛋白酶的例子是商业可获得的(EnzymeDevelopmentCorporation,NewYork,NY,USA)、(EnzymeDevelopmentCorporation,NewYork,NY,USA)和细菌中性蛋白酶(GenencorInternational,Inc.,Rochester,NY,USA)。中性蛋白酶的另一例子是商业可获得的、DSMFoodBeverages,Delft,荷兰的其是从Carica木瓜(一种植物)生产的,即水果来源的酶。
酸性蛋白酶的例子是胃蛋白酶(Sigma,USA)和AcidProtease(AmanoPharma-ceuticalCo.Ltd.,Nagoya,日本)。
在本发明的方法的一种优选实施方式中,随后通过两种不同的碱性蛋白酶,在7至10的pH范围下,40至60℃的温度下,对高粱粉的蛋白质部分进行10至80分钟的处理。
优选地,这些蛋白酶之一是丝氨酸特异性蛋白酶,例如, 或Alkaline另一种是半胱氨酸特异性蛋白酶,例如,或
该步骤还可改变为:不一次加入酶,而是分部分地加入它们(随后或同时)。
步骤c)还可以在步骤d)之后进行,即,先分离高粱蛋白质,然后再对其加以修饰。
步骤d)
优选地,通过离心和/或过滤、优选地通过超滤来进行步骤d)。超滤可以在之前不进行离心的情况下进行。
酶失活后,可以优选地对水解产物进行低速离心(1000g10分钟),以分离不可溶的蛋白质和杂质。
然后可经过Whatman#.4滤纸过滤蛋白质水解产物的可溶级分,并对滤液依次进行超滤(UF)和渗滤(DF),采用截留分子量(MWCO)≥5kDa的膜来进行、优选地采用截留分子量(MWCO)≥30kDa的膜来进行、更优选地采用截留分子量(MWCO)≥50kDa的膜来进行、最优选地采用截留分子量(MWCO)为50至750kDa的膜来进行。
超滤和渗滤可在室温下20至25psi的进口压强和10psi的出口压强下进行。然后可将溶液超滤至浓缩因子为5。超滤后,可立刻用两倍体积的去离子水对滞留物进行两次渗滤。在超滤和渗滤期间,溶液pH可保持为pH8.0至9.0之间,以使得蛋白质保持为可溶状态。
步骤e)
可通过本领域技术人员已知的任何干燥方法来实现向固体形式(例如干粉)的转化。优选的是喷雾干燥或冷冻干燥。喷雾干燥优选地在大约200℃至大约210℃的进口温度和大约70℃至大约75℃的出口温度下进行。冷冻干燥优选地在大约-20℃至大约-50℃的温度下进行10至48小时。
本发明还有一个目的是可通过上文所述的任何方法获得的(经修饰的)高粱蛋白质。
工业应用
本发明涉及上文所述的组合物用于食品、饮料、动物饲料、个人护理组合物和药物组合物的富集、强化和/或着色的用途,以及含有上文所述的组合物的食品、饮料、动物饲料、个人护理组合物和药物组合物本身。
本发明还涉及含有上文所述的(经修饰的)高粱蛋白质的食品、饮料、动物饲料、个人护理组合物和药物组合物,以及此类(经修饰的)高粱蛋白质(优选地,例如上文所述的)作为活性成分(尤其是脂溶性活性成分和/或着色剂)的保护性水胶体的用途。
在本发明的上下文中,动物(包括人)除人之外还尤其包括农场动物,例如绵羊、牛、马、禽类(肉鸡和蛋的色素沉着)、虾和鱼(尤其是鲑鱼和虹鳟鱼)以及宠物,例如猫、狗、鸟(例如,火烈鸟)和鱼。
其中可使用本发明的组合物(尤其是作为着色剂或功能性成分)的饮料可以是碳酸饮料(例如调味苏打水、软饮或矿物饮料)以及非碳酸饮料(例如,调味水、果汁、水果宾治)以及这些饮料的浓缩形式。它们可以基于天然水果或蔬菜汁,或者基于人工香料。其还包括酒精饮料和速溶饮料粉。此外,还包括含糖饮料、无糖饮料(含有无卡路里的人工增甜剂)。
此外,从天然来源获得的或合成的奶制品也在其中可使用本发明的组合物(尤其是作为着色剂或功能性成分)的食物产品的范围内。此类制品的典型例子是奶饮料、冰淇淋、奶酪、酸奶等。奶替代制品,例如豆奶饮料和豆腐制品也包括于此应用的范围内。
本发明还包括含有作为着色剂或功能性成分的本发明组合物的甜品,例如甜食制品、糖果、口香糖、甜点,例如冰淇淋、果冻、布丁、速溶布丁粉等。
本发明还包括含有作为着色剂或功能性成分的本发明组合物的谷类、零食、曲奇、通心粉、汤和酱料、蛋黄酱、沙拉酱等。此外,用于奶制品和谷类的水果制品也包括在内。
基于食物组合物的总重,根据待着色或强化的具体食物产品以及想要着色或强化的程度,经由本发明的组合物而被加入到食物产品中的(脂溶性)活性成分和/或着色剂的最终浓度可以是0.1至500ppm、具体地1至50ppm。
优选地通过以下方法获得本发明的食物组合物:将本发明组合物形式的(脂溶性)活性成分和/或着色剂加入到食物产品中。为对食品或药物产品进行着色或强化,可按照本身已知的用于本发明的水可分散性固体组合物的应用的方法,来使用本发明的组合物。
通常,根据特定的应用,组合物可以作为水性贮液、与其他合适的食品成分的预混合物或干粉混合物来添加。根据最终应用的配方,可以例如使用干粉掺合机、低剪切力混合机、高压均化器或高剪切力混合机来进行混合。显然,此类技术是本领域技术人员已知的。
其中组合物用作着色剂的药物组合物(例如片剂或胶囊)也在本发明的范围内。对片剂的着色可以如下进行:向片剂包衣混合物中以液体或固体着色剂组合物的形式单独加入本发明的组合物,或者将着色组合物加入片剂包衣混合物的一种组分中。着色的硬壳胶囊或软壳胶囊可通过将着色剂组合物掺入胶囊物质的水性溶液来制备。
其中组合物用作活性成分的药物组合物,例如片剂(例如可咀嚼片剂、泡腾片或膜衣片)或胶囊(例如硬壳胶囊),也在本发明的范围内。典型地,本发明的组合物作为粉末加入到压片混合物中,或者以本身已知用于生产胶囊的手段填装进胶囊。
其中组合物用作用于色素沉着(例如用于蛋黄、肉禽、肉鸡或水生动物,尤其是虾、鲑鱼、虹鳟鱼)的着色剂或用作活性成分的动物饲料产品,例如,营养成分的预混物、配合饲料(compoundfeeds)、奶替代品、液体膳食或饲料制剂,也在本发明的范围内。
其中组合物被用作着色剂或用作活性成分的个人护理组合物:化妆品、清洁用品和皮肤产品,即皮肤和头发护理产品,例如,霜、化妆水、浴液、唇膏、香波、护发素、喷雾或凝胶,也在本发明的范围内。
Claims (12)
1.制造经修饰的高粱蛋白质的方法,所述方法包括:
a)制备经碾磨高粱的水性溶液或悬浮液,其中基于所述水性溶液或悬浮液的总量,所述溶液或悬浮液具有0.1至30重量%的干物质含量;
b)任选地移出所述经碾磨高粱的非蛋白质部分或蛋白质部分,来获得高粱蛋白质;
c)修饰经碾磨的高粱的蛋白质部分,来获得经修饰的高粱蛋白质,其中随后通过两种不同的碱性蛋白酶,在7至10的pH范围下,40至60℃的温度下,对高粱粉的蛋白质部分进行10至80分钟的处理;
d)任选地分离经修饰的高粱蛋白质;
e)任选地将经修饰的高粱蛋白质转化为固体形式。
2.如权利要求1所述的方法,其中基于所述水性溶液或悬浮液的总量,所述溶液或悬浮液具有10至15重量%的干物质含量。
3.如权利要求1或2所述的方法,其中在步骤a)、b)和/或c)中可在碾磨之前除去高粱麸。
4.粉末形式的组合物,其包含处于经修饰的高粱蛋白质基质中的一种或多种活性成分,所述经修饰的高粱蛋白质是通过如权利要求1-3中任意一项所述的方法获得的。
5.如权利要求4所述的组合物,其中所述活性成分包含脂溶性活性成分。
6.如权利要求4所述的组合物,其中所述活性成分包含着色剂。
7.如权利要求5所述的组合物,其中所述脂溶性活性成分为胡萝卜素或结构相关的多烯化合物、脂溶性维生素、富含多不饱和脂肪酸的甘油三酯、油溶性UV-A滤光剂、UV-B滤光剂或它们的混合物。
8.如权利要求6所述的组合物,其中所述着色剂为胡萝卜素或结构相关的多烯化合物、脂溶性维生素、富含多不饱和脂肪酸的甘油三酯、油溶性UV-A滤光剂、UV-B滤光剂或它们的混合物。
9.如权利要求7-8中任意一项所述的组合物,其中所述胡萝卜素或结构相关的多烯化合物为类胡萝卜素。
10.如权利要求9所述的组合物,其中类胡萝卜素为α-胡萝卜素、β-胡萝卜素、8’-阿朴-β-胡萝卜醛、8’-阿朴-β-胡萝卜素酸酯、斑蝥黄、虾青素、番茄红素、叶黄素、玉米黄质、藏红花酸、α-玉米胡萝卜素、β-玉米胡萝卜素或它们的混合物。
11.如权利要求9所述的组合物,其中所述类胡萝卜素为β-胡萝卜素。
12.如权利要求7-8中任意一项所述的组合物,其中所述脂溶性维生素为维生素A或维生素E。
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PCT/EP2011/069862 WO2012069322A1 (en) | 2010-11-26 | 2011-11-10 | Protective hydrocolloid for active ingredients |
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CN103636918B (zh) * | 2013-12-13 | 2015-11-04 | 东北农业大学 | 一种富含亚油酸的改性分离蛋白制备方法 |
CN113768028B (zh) * | 2021-09-14 | 2024-01-23 | 辽宁省农业科学院 | 一种从高粱黑粉菌中分离蛋白的方法及其应用 |
CN116121357A (zh) * | 2022-10-14 | 2023-05-16 | 中山大学附属第一医院 | 免疫性肾病标志物及应用 |
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US20130317098A1 (en) | 2013-11-28 |
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AU2011334017A1 (en) | 2013-06-20 |
WO2012069322A1 (en) | 2012-05-31 |
US20140357709A1 (en) | 2014-12-04 |
CN103327827A (zh) | 2013-09-25 |
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