CN104780765B - 用于将谷蛋白降解制备酸面团的微囊化细菌菌群以及该酸面团的制备方法 - Google Patents
用于将谷蛋白降解制备酸面团的微囊化细菌菌群以及该酸面团的制备方法 Download PDFInfo
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Abstract
本发明涉及一种用于谷蛋白降解的微囊化细菌菌群,其包含:(a)市售可得的三种不同的乳酸菌菌株;b)包囊剂;c)益生元;d)海藻糖;再结合一种细菌来源的蛋白水解酶和一种真菌来源的蛋白水解酶。首选微囊化细菌菌群包括:a)植物乳杆菌ATCC 8014;b)旧金山乳杆菌ATCC 27652;c)短乳杆菌ATCC 14869;d)蛋白质含量为90%的分离乳清蛋白;e)葡萄糖值为10的麦芽糖糊精;f)***胶;g)龙舌兰蜜以及h)海藻糖;再结合一种细菌来源的蛋白水解酶和一种真菌来源的蛋白水解酶。本发明还描述微囊化细菌菌群的制备工艺、由微囊化细菌菌群制备酸面团的工艺以及利用上述酸面团制作烘焙食品。
Description
发明领域
本发明涉及制作面包和面包衍生品的食品行业,更具体而言,本发明涉及一种用于酸面团中谷蛋白降解的细菌菌群及其制备工艺,还涉及使用上述无谷蛋白酸面团制作烘焙食品。
发明背景
麦胶性肠病是一种与遗传因素有关、由摄入谷蛋白诱发的免疫***紊乱,谷蛋白是存在于小麦、黑麦和大麦中的一种蛋白质。这种蛋白质在人体上消化道内消化不良。谷蛋白由麦醇溶蛋白和麦谷蛋白两部分组成。麦醇溶蛋白具有醇溶性并包含最多对麦胶性肠病患者有害的成分(Green,P.H.R.和Cellier,C.2007.麦胶性肠病;《新英格兰医学期刊》(N.Engl.J.Med.);357卷:1731-1743)。
麦胶性肠病患者在食用一些含谷蛋白的食物时,他们的免疫***会以损害或破坏肠绒毛的方式做出反应,造成食品的营养成分未被吸收,从而导致营养不良。根据患者的年龄情况,麦胶性肠病的症状可能存在差异,儿童患者多见腹泻、腹胀、呕吐和体重减轻,而成年患者的主要症状是缺铁性贫血、疲劳、骨骼疼痛、关节炎、骨质疏松症等(《美国国家消化道疾病信息交流中心》(National Digestive Diseases Information Clearinghouse),美国国家卫生研究院出版物,2008年第08期:4269)。
最近,曾被认为是罕见疾病的麦胶性肠病成为最常见的不耐症之一,全球每150个新生儿中就有一例麦胶性肠病患者,据估计,只有9%的患者得到确诊。根据墨西哥Salvador Zubirán国家医学科学和营养研究所(Instituto Nacional de Ciencias Médicas y Nutrición)提供的数字,潜在的麦胶性肠病患者有将近260万人(http://celiacosdemexico.org.mx/manifiesto-celiaco)。
目前,麦胶性肠病唯一有效的疗法是在生活中保证无谷蛋白饮食。根据国际食品法典委员的说法,谷蛋白含量小于20ppm的食物被认为是不含谷蛋白的食物。然而,这意味着营养上的负面影响,如减少多糖的摄入,因而降低了能量摄入,减少了对人体健康有益的肠道菌群,从而增加了机会性致病菌的存在。无谷蛋白饮食导致有益菌群减少,对免疫刺激活性产生负面影响并减少抗炎化合物的分泌。
因此,需要提供适合麦胶性肠病患者食用的无谷蛋白产品并相应地增加食品种类以改善患者的饮食。
人类日常饮食中最重要的食品之一是主要使用小麦面粉制作的面包。
面团特性主要取决于谷蛋白蛋白。最近几年,各种不同的处理工艺被应用来提高这些蛋白质的质量或改进制作加气面包的气体保留工艺(Arendt et al.,2007.酸面团对面包口感的影响。《食品微生物学》24:165-174)。这些工艺之一就是使用酸面团制作面包。
这一工艺的主要作用是使面团发酵,使其具有更高的气体含量,从而制作具有柔软面包屑的蓬松面包。
使用面粉(小麦、燕麦、大米等)和水与酵母以及乳酸菌(LAB)的已发酵混合物制作酸面团,其中乳酸菌通常属于乳酸菌属(De Vuyst,L.和Vancanneyt,M.2007.酸面团乳酸菌的生物多样性和识别。《食品微生物学》24.120-127)。酸面团的使用为焙烤工艺提供诸多巨大优势,如发酵过程中pH值降低,气体保持能力更好,抗谷蛋白网络能力更强,抑制面粉淀粉酶,水与谷蛋白及淀粉颗粒结合,戊糖膨胀,植酸盐复合物溶解以及避免发酵不足(DiCagno,R.et al.2002.酸面团乳酸菌蛋白质水解作用:对小麦面粉蛋白以及与人体谷类食物不耐受有关的麦胶蛋白肽的作用。《应用与环境微生物学》(Applied and EnvironmentalMicrobiology)68:623-633)。将这些酸面团与用于制作面包的新鲜面团混合。
此外,面包制作过程中,利用酸面团发酵时使用的乳酸菌(LAB)可实现谷蛋白改性,因此,可实现去除对麦胶性肠病患者有害的蛋白组分。然而,并不是所有的乳酸菌均可将谷蛋白残留含量降低到谷蛋白不耐受者(麦胶性肠病患者)可接受的剂量,因此,需要选择乳酸菌(LAB)类型并找到合适的乳酸菌组合,同时在谷蛋白水解过程中,使用蛋白水解酶以共同发挥作用。
在发酵过程中,乳酸菌(LAB)蛋白酶水解体系释放小分子多肽和氨基酸,可促进微生物的代谢活动,有助于获得更佳口感并减少过敏肽(De Angelis,M.et al.2005.VSL#3“益生菌制剂有能力水解麦醇溶蛋白多肽(脂肪痢益生菌和谷蛋白不耐症的诱因)。”《生物化学与生物物理学报》(Biochimica et Biophysica Acta)1762(1):80-93)的含量,这为对麦醇溶蛋白组分过敏的麦胶性肠病患者带来巨大希望,麦醇溶蛋白是此类患者一直不能摄入即使谷蛋白含量非常少的食品的原因(Wieser,H.2007.“谷蛋白蛋白的化学过程”,《食品微生物学》24:115-119;Di Cagno et al.,2002)。对使用酸面团进行面包制作的过程所做的研究表明:在特定工艺条件下(长时间发酵和半液体),乳酸菌(LAB)可水解小麦醇溶蛋白组分(Di Cagno,R.et al.2004.“脂肪痢患者可耐受用小麦和无毒副作用面粉制作的酸面团面包,开始时进行了乳酸菌选择。”《应用与环境微生物学》70(2):1088-1096.;DeAngelis et al.,2005)。
Rizzello et al.,(“食品加工中,乳酸菌和真菌来源的蛋白水解酶实现高效谷蛋白降解:麦胶性肠病的新视角。”《应用与环境微生物学》73(14):4499-4507,2007)利用酸面团中乳酸菌的非商业菌株(基于其水解麦醇溶蛋白的能力提前选择)和真菌来源的蛋白水解酶的混合物(以其不同组合)在相对较长的发酵过程中去除小麦面粉的毒性。其中还指出乳酸菌水解动力学非常高效,另外,从酸面团中提取的蛋白质诱导了干扰素γ活化;白蛋白、球蛋白、醇溶蛋白完全水解,同时酸面团中依然存在20%的麦谷蛋白。
同样地,美国专利申请第US 2008/0131556号描述了一种由至少六种市售可得的乳酸菌(LAB)和/或双歧杆菌菌种组成的混合物。此混合物可用于酸面团制备。同样,当在这些配方中添加足量的烘焙中常用的细菌来源的蛋白水解酶时,发酵的酸面团中的谷蛋白含量低于200ppm,这可能不适合麦胶性肠病患者食用。同样地,实现这一降解的混合物非常复杂,因为需要使用大量的菌种种类,原因在于使用的乳酸菌(LAB)种类越多,麦醇溶蛋白降解率就越高。
另一方面,国际公布第WO 2010/073283号专利描述了一种包含两种类型乳酸菌(LAB)与一个或多个真菌来源的蛋白水解酶结合的混合物。12小时发酵后,未检测到麦醇溶蛋白和麦谷蛋白。此外,谷蛋白残余含量低于20ppm。
由此可见,在先前的谷蛋白降解技术中已知的细菌培养菌存在某些缺点,例如,使用乳酸菌(LAB)(市售可得的菌株)的复杂混合物(至少六种),或使用特别选择的菌株,则涉及活化、繁殖、清洗并将培养菌悬液(菌种)添加至面团中,需要每天制备它们,且需要高度专业化的管理来避免污染,以便使培养菌在同一发育阶段保持活性,同时应在乳酸菌(LAB)菌种之间保持适当的配比、充足数量,这意味着存在潜在的污染风险。
另一种降低与每次制备菌种有关的一系列难度的方法是将培养菌微囊化,这样可制备足够量的培养菌,即可减少培养菌操作,从而降低与污染及乳酸培养菌活力相关的风险,并提高面团降解的工艺效率。
乳酸菌(LAB)活力和活性由一系列因素决定,包括培养菌增殖速度、乳酸和醋酸的产酸能力、总固体含量、培养温度及时间,所使用菌种的数量以及抗生素、消毒剂或洗涤剂残留物数量(Picot与Lacroix,2003;2005;Desmond et al.,2002;Favaro-Trinidad and Grosso,2002;Lian et al.,2003;Akalin et al.,2004;Cerdeira et al.,2005;Iyer与Kailasapathy,2005;Ozer et al.,2005;Ann et al.,2007)。
存在一些不同的提高益生元抗逆性的方法,如微囊化以及将微营养素和益生元功能结合在一起(Picot与Lacroix,2003;Chandramouli et al.,2004;Talwalkar与Kailasapathy,2004;Cerdeira et al.,2005;Iyer与Kailasapathy,2005;Ann et al.,2007)。
由于益生元对高浓度的氧气、生产、存储、冷冻、通过胃肠道过程中的酸碱性条件具有高度敏感性,微胶囊技术被建议用来增加益生元的活力(Favaro-Trinidad与Grosso,2002;Picot与Lacroix,2003;Chandramouli et al.,2004;Iyer与Kailasapathy,2005;Crittenden et al.,2006;Ann et al.,2007)。
尽管乳酸菌(LAB)的微胶囊技术在现有技术中已逐渐被人们熟知,微胶囊技术可确保为待微囊化的物质提供良好的保护,而且在酸面团发酵期间将用于降解谷蛋白,但是选择良好的干燥条件和优良的胶囊介质至关重要,在现有技术中未对胶囊介质做出描述,因为微胶囊囊壁设计是一个关键性问题,其取决于待保护物质的特定类型、微胶囊的应用环境以及微胶囊所保护的活性物质的释放机制。
发明目的
考虑到先前技术的缺陷,本发明的目的之一是提供一种微囊化的乳酸菌(LAB)菌株的微生物菌群,其结合一种细菌来源的蛋白水解酶和一种真菌来源的蛋白水解酶,适于制备酸面团,另外,其十分稳定,可快速活化且易于操作,可直接添加至小麦面团中降解谷蛋白,包括降解对麦胶性肠病患者有害的组分。
此外,本发明的另一个目标是使用上述微囊化的微生物菌群制备酸面团,其中,谷蛋白得到降解,其工艺非常简单并且可用于工业规模。
本发明的另外一个目标是使用酸面团制备适合麦胶性肠病患者食用的不含谷蛋白的烘焙食品。
发明概述
为此,发明了一种用于降解谷蛋白的微囊化细菌菌群,其包含:(a)市售可得的三种不同的乳酸菌菌株;b)包囊剂;c)益生元;d)海藻糖;再结合一种细菌来源的蛋白水解酶和一种真菌来源的蛋白水解酶。
本发明的其他方面考虑用于谷蛋白降解的微囊化细菌菌群的制备工艺;利用上述细菌菌群结合细菌来源的蛋白水解酶和真菌来源的蛋白水解酶制作的酸面团,其中谷蛋白含量低于20ppm;该酸面团的制备方法以及由该酸面团制作的不含谷蛋白的烘焙食品。
发明说明
本发明研究期间,意外发现一种微囊化细菌菌群在结合使用包囊剂、益生元、海藻糖、细菌来源的蛋白水解酶和真菌来源的蛋白水解酶时,可在酸面团发酵过程中用于谷蛋白降解。其中,这种微囊化细菌菌群由三种不同的市售可得的乳酸菌菌株组成;包囊剂可为细菌菌群提供高水平保护,该包囊剂可在面团***中快速溶解,因此可迅速开始其活力;益生元可让菌群达到最高存活率;海藻糖作为细菌的特定营养成分,而细菌来源的蛋白水解酶和真菌来源的蛋白水解酶两者均可市场上可买到。
首选的乳酸菌菌株是植物乳杆菌ATCC 8014,旧金山乳杆菌ATCC 27652以及短乳杆菌ATCC 14869。
包囊剂可从一组制剂中选择,具体包括:蛋白质含量为90%的分离乳清蛋白、葡萄糖值为10的麦芽糖糊精、***胶、牧豆树胶、海藻酸钠、果胶、大豆分离蛋白、酪蛋白及其任意组合,首选蛋白质含量为90%的分离乳清蛋白、葡萄糖值为10的麦芽糖糊精、***胶及其任意组合。
另一方面,益生元可以是聚葡萄糖、菊粉和龙舌兰蜜,首选龙舌兰蜜。
细菌来源的蛋白水解酶和真菌来源的蛋白水解酶可以是烘焙常用酶。
在本发明的一个具体实施例中,微囊化细菌菌群包含:a)植物乳杆菌ATCC 8014;b)旧金山乳杆菌ATCC 27652;c)短乳杆菌ATCC 14869;d)蛋白质含量为90%的分离乳清蛋白;e)葡萄糖值为10的麦芽糖糊精;f)***胶;g)龙舌兰蜜以及h)海藻糖;再结合一种细菌来源的蛋白水解酶和一种真菌来源的蛋白水解酶。
更可取地,该微囊化细菌菌群包括:a)植物乳杆菌ATCC 8014;b)旧金山乳杆菌ATCC 27652;c)短乳杆菌ATCC 14869;d)40%至60%的蛋白质含量为90%的分离乳清蛋白;e)10%至20%的葡萄糖值为10的麦芽糖糊精;f)20%至80%的***胶;g)1%至10%的龙舌兰蜜以及h)0.5%-5%的海藻糖;再结合一种0.005%至0.03%的细菌来源的蛋白水解酶和一种0.001%至0.02%的真菌来源的蛋白水解酶。
微胶囊技术采用现有技术中的众所周知的工艺,如喷雾干燥。例如,可按照如下步骤制备微囊化细菌菌群:
a)分别再活化每株乳酸菌;
b)一旦每个菌株均活化,在液体培养基内分别进行培养,直至每个菌株达到所需的浓度;
c)移除每个菌株中多余的培养基,以浓缩微生物,优选通过离心法,获得沉淀物;
d)分别在盐水悬液中重新悬浮每个菌株获得的沉淀物并调整至所需体积;
e)混合所需数量的每种菌株,并调至最后体积;
f)以适当比例将包囊剂溶解在水中,以使得具有20%至30%的溶解性固体;
g)添加益生元和海藻糖;
h)接种此三株乳酸菌的混合物,以得到粉末状微囊化的上述三株乳酸菌混合物,每克浓度约为1010CFU;
i)干燥,优选采用喷雾干燥法,入口温度介于110℃至160℃之间,出口温度介于60℃至80℃之间,进给速率介于20mL/min至50mL/min之间。
作为更可取的方案,可在步骤e)中以10%-50%比率的植物乳杆菌ATCC 8014混合是、40%比率的旧金山乳杆菌ATCC 27652,10%-50%比率的短乳杆菌ATCC 14869进行混合。
最好采用喷雾干燥器进行干燥处理,入口温度150℃,出口温度80℃,进给速率22mL/min。
本发明的另一个方面考虑一种使用本发明的微囊化细菌菌群结合细菌来源的蛋白水解酶和真菌来源的蛋白水解酶制作的酸面团,其中,该酸面团的谷蛋白含量低于20ppm,最佳条件下可低于10ppm,适合制作不含谷蛋白的烘焙食品。
若要在酸面团中获得这一谷蛋白含量,最好发酵三个小时以上,最佳发酵时间为31小时以上。
本发明的酸面团制备工艺包括如下步骤:
1)将30%至60%的面粉与40%至80%的水混合,制成面团得率介于150至475之间的面团,优选面团得率为150至160;
2)添加提前溶解于和面用水中的0.005%至0.03%的细菌来源的蛋白水解酶和0.001%至0.02%的真菌来源的蛋白水解酶;
3)加入微囊化细菌菌群,使得每克面粉中含有约1010CFU三株乳酸菌混合物;
4)混合;和
5)发酵3至48个小时,优选发酵28至35个小时,温度为30至37℃,相对湿度为70%至92%,优选相对湿度为75%至80%。
步骤1)中使用的面粉可以是小麦面粉、黑麦面粉和燕麦面粉,首选小麦面粉。
关于发酵,最好在温度35℃、相对湿度76%的条件下发酵31小时。
如上所述,本发明中以上述工艺制备的酸面团的谷蛋白含量低于20ppm,最佳条件下谷蛋白含量可低于10ppm。若要在酸面团中获得这一谷蛋白含量,最好发酵三个小时以上。
使用这些酸面团可制作烘焙食品,特别适宜制作适合麦胶性肠病患者食用的不含谷蛋白甜面包。
通过以下示例可更好地理解本发明,这些示例仅用作说明用途,以实现对本发明优选实施例的正确理解,并非意味着不能基于上文的详细描述实施其他非说明性实施例。
示例
示例一
菌种制备
从美国菌种保藏中心(ATCC)的一个供应商处获得植物乳杆菌ATCC 8014、短乳杆菌ATCC 14869和旧金山乳杆菌ATCC 27652三种菌株,前两种菌株保存于固体培养基中,对第三种菌株进行冻干处理。
为了将菌株再活化,将包含于Man Rogosa Sharpe(MRS)液体培养基(DifcoTM,USA)中的菌种放置于带螺旋盖的试管内,以35至37℃的温度培养24至48个小时,或者观察其生长,直至浊度增加。
当培养菌显现这一特征时,将它们转移至有盖培养皿内的MRS固体培养基中,以相同的温度再培养24至48个小时。
一旦微生物长成,再将它们分别转移至含有50ml MRS培养基的125mL锥形烧瓶中,以37℃的温度培养8至12个小时,同时以50至100rpm的转速搅拌。此即为预制菌种。
经过上述培养时间之后,再次在无菌条件将该培养菌转移至含有500至1000mlMRS培养基的2800mL冯巴赫瓶(Fembach flask)中,以37℃的温度培养10至16个小时以上,同时以50至150rpm的转速搅拌。对每个烧瓶进行取样,以测定培养基吸光度(浊度)。
为了移除剩余的培养基并对微生物进行浓缩,将该培养菌放置于经过酒精消毒的250mL离心瓶中,随后以2000至5000rpm的转速进行离心机离心处理,时长20分钟。
将获得的沉淀物再悬浮于无菌生理盐水中(氯化钠,9g/L)。一旦微生物完成再悬浮且已调整至所需的体积,依然在无菌条件下将所需数量的每种菌株混合在一起并达到最终体积,以此制备待微囊化的菌种。
示例二
细菌菌群的微囊化
蛋白含量90%的乳酸分离乳清蛋白(具备氧气阻隔性能)、葡萄糖值为10的麦芽糖糊精以及***胶被选作包囊剂(以不同配比)。
为支持菌群中细菌的再活化,对添加的三种类型的益生元制剂进行分析:聚葡萄糖、菊粉及龙舌兰蜜。而且,海藻糖也被用作特定的营养成为,帮助细菌形成菌群。
以适当比例将包囊剂溶解在水中,其中含有20%至30%的溶解性固体。添加益生元和海藻糖,接着为该分散体接种适量的细菌菌群,以此获得1010CFU/g粉末状微胶囊。
使用喷雾干燥器对之前的混合物进行干燥处理,入口温度150℃,出口温度80℃,进给速率22mL/min。
因此,即获得了粉末状微囊化的微生物菌群。
示例三
益生元配比测定有助于乳酸菌(BAL)发育
如上所述,对添加至微囊化菌群中的聚葡萄糖(P)、菊粉(I)、龙舌兰蜜(M)进行计算。为此,在实验中采用了单一质心混合物设计,如表1所示:
表1、用于微生物菌群微囊化的益生元配方计算
将每种混合物添加至相应乳酸菌(LAB)分散体和壁材中,随后通过上述的喷雾干燥法进行干燥处理。
一旦获得微胶囊,取1g该粉末,使用无菌水将其再水化,至30℃,同时搅拌。取1ml试样,对其进行系列稀释,并实施平板接种,以测定最大可能数。细菌存活率结果如表2所示。
表2、乳酸菌(LAB)存活率取决于益生元组合
根据以上数据,可见当益生元为龙舌兰蜜时可获得最大存活率(98%)。
示例四
酸面团制备
采用市场上可买到、适于手工烘焙的白面粉以及根据示例二获得的微囊化细菌菌群(浓度为1010CFU/g),除了包囊剂和海藻糖之外,还包含龙舌兰蜜作为益生元。
至于蛋白水解酶,采用一种细菌来源的蛋白水解酶(HT蛋白水解200,EnmexS.A.de C.V.,墨西哥)和另一种真菌来源的蛋白水解酶(Harizyme G,Enmex S.A.de C.V.,墨西哥)。
就酸面团制备而言,使用小麦面粉制备MY(面团得率)介于150与160之间的面团,混合400g面粉和150mL水。添加0.08g细菌来源的蛋白水解酶(HT蛋白水解200,EnmexS.A.de C.V.,墨西哥)和0.12g真菌来源的蛋白水解酶(Harizyme G,Enmex S.A.de C.V.,墨西哥),两种蛋白水解酶均提前溶解于和面用水中。接着,加入本发明的微生物菌群微胶囊,悬浮于水中,以此获得1010CFU/g浓度。
在标准和面机中揉搓8分钟,然后在温度35℃、相对湿度76%的条件下发酵31小时。通过电泳分析(西方墨点法)监测发酵期间的pH值以及发酵结束时的谷蛋白降解强度。结果如表3所示。
表3、接种微囊化细菌菌群的酸面团的降解动力学
根据上表所示的数据,可以看到在发酵的最初三个小时之内在本发明的微囊化细菌菌群的影响之下,再结合蛋白水解酶的作用,谷蛋白醇溶蛋白发生密集降解。
示例五
使用经乳酸发酵的酸面团精心制作阿斯托加(Astorga)杯形甜面包
使用根据示例四制备的酸面团制作不含谷蛋白的杯形甜面包。为此,使用搅拌混匀器将黄油、人造奶油和糖玻璃搅打至糊状,起初低速搅打直至打发状态,随后增加搅打速度,继续搅拌20分钟,将最大量的空气掺入该混合物中。
将三个蛋黄逐步加入其中,直至其完全打发,随后加入牛奶、调味品(鲜橙、柠檬和巧克力)和山梨酸钾。
将根据示例四制备的发酵酸面团、玉米淀粉、经碱液处理(nixtamalized)的面粉、小苏打和发酵粉在塑料袋中预混合之后,掺入之前的混合物中。加入水,随后高速搅拌8分钟。
将蛋白单独搅打至硬身全发,随后将其与其余的面团混合在一起,以中速混合,以免破坏泡沫。
将液态混料倒入杯形纸杯中,放入烤箱烤盘中,以200℃烘焙10分钟。
示例六
消费者评估阿斯托加(Astorga)杯形蛋糕
为了确定根据示例五(使用微囊化微生物菌群并结合一些蛋白水解酶制备的酸面团)制作的鲜橙、柠檬和巧克力口味的杯形蛋糕相对于采用传统面团制作的杯形蛋糕的受欢迎度,于是面向消费者展开了一项评估。
为此,对伊比利亚美洲大学(Universidad Iberoamericana)墨西哥城校园的70位阿斯托加(Astorga)杯形蛋糕消费者展开了一项测试。参与者中男女比例各占一半,年龄从15岁至45岁以上不等,这些参与者是甜面包食品的消费者,至少每15天食用一次甜面包。该项测试的显著性和置信水平分别是0.05%和95%。结果如表4所示。
表4、采用含有微囊化细菌菌群(结合蛋白酶)的发酵酸面团制成的杯形蛋糕的消费者测试结果
表4中的结果表明,就原味杯形蛋糕而言,在95%的置信水平下所有的评估特征(卖相、口感和口味)不存在显著性差异(显著性仅0.05%),而且对于鲜橙口味和巧克力口味杯形蛋糕的总体接受度也不存在显著性差异。
相反,相比原味杯形蛋糕而言,在95%的置信水平下柠檬口味杯形蛋糕的确在所有的评估特征上呈现显著性差异,消费者更偏爱原味杯形蛋糕。
按照上述内容,可以看到用于酸面团中谷蛋白降解的微囊化细菌菌群和该酸面团的制备工艺,以及使用这种酸面团制作的烘焙制品均设计用于制作可能适合麦胶性肠病患者食用的不含谷蛋白食品。本领域的技术人员显然会认识到,如前所述,用于酸面团中谷蛋白降解的微囊化细菌菌群以及酸面团制备工艺的所述实施例仅为本发明的说明性、非限制性实施例,可以在不脱离本发明范围的情况下对其细节进行各种变更考虑。
因此,除先前技术所要求的限制以及所附权利要求中所指出的限制之外,本发明不应被视为限制性的。
Claims (24)
1.一种用于谷蛋白降解的微囊化细菌菌群,其特征在于该微囊化细菌菌群包括:(a)以下三种不同种类的乳酸菌:植物乳杆菌ATCC 8014、旧金山乳杆菌ATCC 27652和短乳杆菌ATCC 14869;b)包囊剂;c)益生元;和d)海藻糖;
该微囊化细菌菌群与细菌来源的蛋白水解酶和真菌来源的蛋白水解酶组合。
2.根据权利要求1所述的微囊化细菌菌群,其中,所述包囊剂选自:蛋白质含量为90%的分离乳清蛋白、葡萄糖值为10的麦芽糖糊精、***胶、牧豆树胶、海藻酸钠、果胶、大豆分离蛋白、酪蛋白及其组合。
3.根据权利要求2所述的微囊化细菌菌群,其中,所述包囊剂是蛋白质含量为90%的分离乳清蛋白、葡萄糖值为10的麦芽糖糊精、***胶及其组合。
4.根据权利要求1所述的微囊化细菌菌群,其中,所述益生元选自聚葡萄糖、菊粉和龙舌兰蜜。
5.根据权利要求4所述的微囊化细菌菌群,其中,所述益生元是龙舌兰蜜。
6.根据权利要求1所述的微囊化细菌菌群,其中,所述细菌来源的蛋白水解酶和真菌来源的蛋白水解酶选自烘焙常用酶。
7.根据权利要求1所述的微囊化细菌菌群,其包括:a)植物乳杆菌ATCC8014;b)旧金山乳杆菌ATCC 27652;c)短乳杆菌ATCC 14869;d)蛋白质含量为90%的分离乳清蛋白;e)葡萄糖值为10的麦芽糖糊精;f)***胶;g)龙舌兰蜜以及h)海藻糖;
该微囊化细菌菌群与细菌来源的蛋白水解酶和真菌来源的蛋白水解酶组合。
8.根据权利要求7所述的微囊化细菌菌群,其包括:a)植物乳杆菌ATCC8014;b)旧金山乳杆菌ATCC 27652;c)短乳杆菌ATCC 14869;d)40%至60%的蛋白质含量为90%的分离乳清蛋白;e)10%至20%的葡萄糖值为10的麦芽糖糊精;f)20%至80%的***胶;g)1%至10%的龙舌兰蜜以及h)海藻糖;
该微囊化细菌菌群与0.005%至0.03%的细菌来源的蛋白水解酶和0.001%至0.02%的真菌来源的蛋白水解酶组合。
9.一种获得权利要求1所述的用于谷蛋白降解的微囊化细菌菌群的制备工艺,其特征在于该制备工艺包括:
a)分别再活化每株乳酸菌;
b)一旦每个菌株均活化,在液体培养基内分别进行培养,直至每个菌株达到所需的浓度;
c)移除每个菌株中多余的培养基,以浓缩微生物,获得沉淀物;
d)分别在盐水悬液中重新悬浮每个菌株获得的沉淀物并调整至所需体积;
e)混合所需量的每种菌株,并调至最后体积;
f)以适当比例将包囊剂溶解在水中,以使得具有20%至30%的溶解固体;
g)添加益生元和海藻糖;
h)接种所述三株乳酸菌的混合物,以得到粉末状微囊化的上述三株乳酸菌混合物,每克浓度为1010CFU;和
i)喷雾干燥,入口温度介于110℃至160℃之间,出口温度介于60℃至80℃之间,进给速率介于20mL/min至50mL/min之间。
10.根据权利要求9所述的微囊化细菌菌群的制备工艺,其中通过离心法移除多余的培养基。
11.根据权利要求9所述的微囊化细菌菌群的制备工艺,其中,采用喷雾干燥器进行喷雾干燥处理,入口温度150℃,出口温度80℃,进给速率22mL/min。
12.一种由权利要求1所述的微囊化细菌菌群结合细菌来源的蛋白水解酶和真菌来源的蛋白水解酶而制备的酸面团,其特征在于具有低于20ppm的谷蛋白浓度。
13.根据权利要求12所述的酸面团,其具有低于10ppm的谷蛋白浓度。
14.根据权利要求12所述的酸面团,其中,经过3小时发酵之后获得所述低于20ppm的谷蛋白浓度。
15.根据权利要求12所述的酸面团,其中在31小时发酵后获得所述低于20ppm的谷蛋白浓度。
16.一种由权利要求1所述的用于谷蛋白降解的微囊化细菌菌群结合细菌来源的蛋白水解酶和真菌来源的蛋白水解酶来制备酸面团的工艺,其包括以下步骤:
i)将30%至60%的面粉与40%至80%的水混合,制成面团得率介于150至475之间的面团;
ii)添加提前溶解于和面用水中的0.005%至0.03%的细菌来源的蛋白水解酶和0.001%至0.02%的真菌来源的蛋白水解酶;
iii)加入微囊化细菌菌群,使得每克面粉中的含量为1010CFU;
iv)混合;和
v)发酵3至48个小时,温度为30至37℃,相对湿度为70%至92%。
17.根据权利要求16所述的酸面团制备工艺,其中在步骤i)中的面团得率为150至160。
18.根据权利要求16所述的酸面团制备工艺,其中步骤i)中使用的面粉选自小麦面粉、黑麦面粉和燕麦面粉。
19.根据权利要求18所述的酸面团制备工艺,其中所述面粉是小麦面粉。
20.根据权利要求16所述的酸面团制备工艺,其中步骤v)的发酵进行28至35个小时。
21.根据权利要求16所述的酸面团制备工艺,其中步骤v)的发酵在相对湿度75%至80%下进行。
22.根据权利要求16所述的酸面团制备工艺,其中所述发酵是在温度35℃、相对湿度76%的条件下发酵31小时。
23.由权利要求12所述的酸面团制成的烘焙产品,其不含谷蛋白。
24.根据权利要求23所述的烘焙产品,其为甜面包。
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