CN113710102A - 为人、动物和农业目的提高天然复合多糖的糖的生物利用度的生产方法 - Google Patents
为人、动物和农业目的提高天然复合多糖的糖的生物利用度的生产方法 Download PDFInfo
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- CN113710102A CN113710102A CN202080022912.2A CN202080022912A CN113710102A CN 113710102 A CN113710102 A CN 113710102A CN 202080022912 A CN202080022912 A CN 202080022912A CN 113710102 A CN113710102 A CN 113710102A
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Abstract
公开了包含由益生菌共混物发酵的植物底物的发酵食品或食品补充剂。根据本发明的发酵食品或食品补充剂特别适用于改善自然母性、哺乳和婴儿断奶。本发明还公开了一种制备发酵食品或食品补充剂的方法,包括用益生菌共混物发酵植物底物。
Description
技术领域
本发明涉及发酵食品或食品补充剂及其制备方法。
发明背景
几个世纪以来,发酵食品在世界各地传统上用于人类营养。不同类型的食物被用作发酵工艺的底物,包括蔬菜、肉、奶、鱼等。基于这种底物在该生态位中的普遍可用性,生态环境强烈影响要发酵的底物类型。
已知发酵可以提高原始底物的营养价值以及延长所谓的食品保质期,使其即使在恶劣的环境条件下也可以长时间储存。
发酵工艺(特别是对于传统食品而言)基于细菌、真菌和酵母的自发群落的活动,这些活动一起,使细菌、真菌和酵母从底物中获利的能力最大化。一些天然微生物发酵群落代表了基因不同的微生物之间共生的极好例子。不幸的是,微生物的自然自发发酵池必须保持连续性以维持其特征,但与其发酵活动相关的代谢物和风味的产生无法控制。然而,在大多数情况下,由于监管和安全要求,这种不受控制的工艺不适用于发酵食品的工业生产。
发酵食品历来是农村社区生计以及当地传统手工生产的宝贵资源。除了作为这种中小型经济的创收工具外,发酵确保了更强的安全性要求,即使自然资源季节性变化,也允许设计适合长期储存的非常多样化的加工食品模式。(FAO,2012)。
通常发酵食品来自于由细菌、酵母、真菌和霉菌的自然群落共同作用而进行的自然和自发的食物降解过程,以充分利用底物的代谢优势。自发发酵群落的传统生产工艺通常通过文化和传统价值传递给后代。
基于谷物、水果和蔬菜的发酵食品以及发酵酒精饮料的广泛传播证明了发酵食品与人类营养的相关性。FAO对1998年至2000年世界上最著名的发酵食品进行了详细而全面的概述。
推广发酵食品的主要原因是由于底物的酸化确保了其保质期的延长,但其他好处通常与这一传统技术优势相关,其在农村社区很早开始的发酵食品的发展中得到推广。除了发酵的防腐作用外,其例如通过降低抗营养素的浓度还为营养底物提供适口性以及提高其营养价值(Selhub等人,2014年)。
传统上,通过食物与人体肠道微生物群相互作用的发酵食品以及改善肠道功能对消费者福祉的积极影响也对健康有益。然而,这些有益作用背后的机制尚未阐明。
自然发酵的微生物群落由细菌、真菌、霉菌和酵母的复合群体组成,它们有些可培养,有些不可培养,并能够利用不同的食物底物(Tamang等人,2016年)。许多研究项目都集中在对这些自然自发群落组成的理解上,世界各地的各种食品和饮料的自然自发群落组成有很大差异。无论如何,基于分类标准对不同成分的可靠识别始终是一个非常困难的问题,导致对这些生态群体的了解不完整。
适当控制的发酵可以确保有益营养素的最大化并减少由于抗营养素和污染物引起的潜在有害影响,放大发酵食品的除了营养支持作用外的有益作用。
选择用于食品受控发酵的发酵剂菌株的当前方法侧重于实现最佳发酵性能(例如用于发酵面包店面粉的酵母)和/或降低污染风险(例如用于肉类的发酵剂乳酸菌)和/或促进特殊且典型的风味和代谢物的研发(例如酸奶、葡萄酒等的发酵剂菌株)。
另一方面,益生菌是活的微生物,当给予足够的量时,能够对宿主产生有益的影响(FAO,2002)。具有经证实的有益特性的微生物菌株可作为食物(例如发酵乳)以及食品补充剂甚至药物供人类食用。发挥其有益作用的主要和必要特征是,它们必须以有活力的形式被消耗,并通过肠道通道保持其活力。无论如何,专家认为在某些危急情况下使用益生菌似乎是有问题的。
通过进行能够在对特殊人群的有益效果方面最大限度地发挥底物特性的受控发酵,精心挑选的益生菌可以与牛奶以外的不同食物底物相关联。孕妇、哺乳期的母亲、断奶期间的婴儿通常经历内源性缺乏双歧杆菌,双歧杆菌是一组关键的细菌,其有益功能主要依赖于产生结肠功能所必需的短链脂肪酸。为了促进双歧杆菌的选择性发育,一些益生元,例如低聚果糖和低聚半乳糖被外源添加到断奶食品中(Sholtens等人,2006年)。
选择的发酵剂菌株是在工业水平上开发的,以促进食品的受控发酵。然而,通常因为发酵剂菌株的技术特征例如因为它们能够快速降解底物,而不是因为它们对消费者的潜在益处和/或它们提高食物营养价值的能力而选择发酵剂菌株。
发明内容
现在已经发现,通过合适选择特异性细菌共混物和特异性发酵条件,可以获得食品和食品补充剂的新发酵成分,所述食品和食品补充剂通过来自微生物活动的有用代谢物的积累以及原始食物底物中存在的抗营养素的减少例如谷物和豆类中植酸的减少,为宿主提供有益的作用。特别地,已经发现了由格氏乳杆菌(Lactobacillus gasseri)、鼠李糖乳杆菌(Lactobacillus rhamnosus)和短双歧杆菌(Bifidobacterium breve)组成的特异性选择的细菌共混物,能够有效地发酵植物源食品(food of vegetable origin)。
因此,本发明的第一个目的包括发酵食品或食品补充剂,其包含通过由格氏乳杆菌、鼠李糖乳杆菌和短双歧杆菌组成的益生菌共混物发酵的植物底物(vegetablesubstrate)。
本发明的第二个目的包括制备发酵食品或食品补充剂的方法,该方法包括植物底物与鼠李糖乳杆菌的第一发酵步骤,随后是格氏乳杆菌和短双歧杆菌的第二发酵步骤。
本发明还涉及所获得的发酵食品或食品补充剂用于改善自然母性、哺乳和婴儿断奶的用途。
具体实施方式
根据本发明,可以以任何比例使用格氏乳杆菌、鼠李糖乳杆菌和短双歧杆菌的任何菌株。以细胞计数表示的1:1:1的比例是优选的。
优选菌株已根据布达佩斯条约以下列登录号保藏在根特大学(GhentUniversity)(比利时)的BCCM/LMG保藏中心:
格氏乳杆菌(Lactobacillus gasseri)LMG P-30998
鼠李糖乳杆菌(Lactobacillus rhamnosus)LMG P-31000
短双歧杆菌(Bifidobacterium breve)LMG P-30999
通过根据本发明的益生菌共混物进行发酵的植物底物可以选自燕麦、大麦、玉米、谷物、假谷物、无麸质蔬菜、蔬菜、水果、坚果,任选地与例如玉米油、葵花籽油或橄榄油等油组合。燕麦特别优选作为底物,单独或与玉米油、葵花籽油或其他植物油组合。
如果需要,本发明的发酵食品或食品补充剂还可含有动物奶,优选驴奶或绵羊奶。
本发明的发酵食品可以是任何适合施用的形式,例如液体、粉末或奶油形式。所述食品可以作为食品成分和/或食品补充剂,以0.5-20重量%的浓度添加到常规食品中,如酸奶、牛奶、配方奶、果汁、软饮料、婴儿食品和果酱等。
用于植物底物发酵的益生菌共混物在本发明的食品或食品补充剂中是没有活力的,本发明的食品或食品补充剂经过巴氏杀菌工艺以确保在生产工艺结束时其不含有有活力的微生物细胞。根据最终加工条件,所得成分具有不同的组成和特征,取决于某些微生物代谢物和分解代谢/合成代谢产物的失活和/或保存情况。
本发明的发酵食品或食品补充剂通过以下方法制备,包括植物底物与鼠李糖乳杆菌的第一发酵步骤,然后是使用格氏乳杆菌和短双歧杆菌的第二发酵步骤。事实上,由于双歧杆菌无法使用燕麦饮料作为生长培养基,并且仅观察到格氏乳杆菌的部分性能,因此发酵是通过先用鼠李糖乳杆菌接种培养基,然后在校正pH至6.0之后添加格氏乳杆菌和短双歧杆菌来进行。通过这种策略(两步发酵),底物首先被鼠李糖乳杆菌降解,产生短链糖及游离肽和氨基酸,能够更好地支持双歧杆菌的生长。
发酵一旦完成,立即对底物进行热处理以灭活细菌。灭活的发酵产物可直接用于动物或人类食用。否则,产品将通过不同的技术方法制成粉末,例如通过冻干、喷雾干燥或类似技术。
在冻干的情况下,将灭活的发酵底物冷冻至-20℃-80℃,然后在冻干机中干燥。定时和干燥条件取决于设备的大小。最终粉末的水分含量为3.0%至4.5%。获得白色且具有甜味的细粉末。从液体发酵底物中回收粉末的百分比为10%至30%。
在喷雾干燥的情况下,灭活的发酵底物被直接进料到喷雾干燥器并在不同的入口和出口温度(80至180℃)和不同的流量(以ml/min计)下处理。最终粉末的水分含量为2.0%至4.0%。粉末外观细腻,为白色至微黄色,味甜。从液体发酵底物中回收粉末的百分比为20%至60%。为设定最佳生产条件而进行的分析表明,较低的喷雾温度产生较高的干燥粉末的回收率。
两步发酵工艺允许通过积累有用的代谢物、降低潜在危险物质的浓度和提高天然食物,特别是蔬菜的消化率,最大化所选共混物的能力,以提高原始食物底物的质量。
本发明的发酵食品提供了许多有益效果,包括刺激双歧杆菌、减少抗营养素如植酸、改善自然母性、哺乳和婴儿断奶,通过作用于人体肠道黏膜的内源性大麻素系统来调节腹痛,促进天然植物性食物的消化率和增加某些营养素的生物利用度。通过促进双歧杆菌的生长,从而改善肠道微生物群的组成,本发明的发酵食品有效地对抗病原菌或潜在有害微生物。本发明因此丰富和改善了食物选择非常有限的特殊类别的消费者例如断奶婴儿和老年人的饮食。
新发酵食品对自然改善母性、哺乳和断奶的功效通过几项体外测试进行评价,这些测试最终通过直接接种牛奶和粪便发酵模型(FMF)测定双歧杆菌的功效(作为益生元物质的能力,确定双歧杆菌的增加),从而通过作用于人体肠道黏膜的内源性大麻素系统来调节腹痛,并减少天然植物源基质中含有的抗营养素含量,促进天然植物食品的消化率并提高营养素的生物利用度。所得结果在以下实施例中报告。
实施例1-发酵食品的制备
根据本发明的工艺,通过首先用鼠李糖乳杆菌进行发酵,然后用上述两种其他细菌物种进行发酵(两步发酵),或者通过在与发酵1报道的相同条件下同时使用三种细菌物种发酵底物,使用并比较了所获得的不同菌株的以下共混物。
共混物A
格氏乳杆菌L6
鼠李糖乳杆菌L13b
短双歧杆菌2TA
共混物B
格氏乳杆菌20243
鼠李糖乳杆菌GG
短双歧杆菌BB03
共混物C
格氏乳杆菌LG36
鼠李糖乳杆菌Sp1
短双歧杆菌M16V
母培养物的制备:通过在培养基中培养三种细菌物种、离心以除去用过的上清液、用纯水洗涤并用新鲜培养基或纯水重悬,制备构成共混物的3种菌株的洗涤纯培养物。在平板上计数母培养物以检查活力。
第1发酵(一步发酵):用最终浓度为1%的鼠李糖乳杆菌、格氏乳杆菌和短双歧杆菌清洗的培养物(10^5-10^8CFU/ml的培养基)来接种食物底物。
在微需氧条件、静态或缓慢搅拌下,在37℃下温育18-24小时。
活的鼠李糖乳杆菌、格氏乳杆菌和短双歧杆菌的最终计数各自为至少3x10^8CFU/ml,pH为3至4。
通过热处理例如在65℃x30分钟下巴氏杀菌灭活细菌的活细胞。
通过氢氧化钠(碳酸氢钠和/或其他碱性试剂也适用)将pH值校正为6.0-7.0。
第2发酵(两步发酵)
第1阶段-用1%终浓度的鼠李糖乳杆菌洗涤的培养物(10^5-10^8CFU/ml的培养基)接种食物底物。
在微需氧条件下、静态或缓慢搅拌下,在37℃下温育18-24小时。
活鼠李糖乳杆菌的最终计数各自为至少3x10^8CFU/ml,pH为3至4。
通过热处理例如在65℃x30分钟下的巴氏杀菌灭活细菌的活细胞。
通过氢氧化钠(碳酸氢钠和/或其他碱性试剂也适用)将pH校正为6.0-7.0。
第2阶段-用1%终浓度的格氏乳杆菌和短双歧杆菌洗涤的培养物(10^5-10^8CFU/ml的培养基)接种在第1阶段发酵的底物。
在微需氧条件、静态或缓慢搅拌下,在37℃下温育18-24小时。
活格氏乳杆菌和短双歧杆菌的最终计数各自为至少3x10^8CFU/ml,pH为3至4。
通过热处理例如在65℃x30分钟下的巴氏杀菌灭活细菌的活细胞。
通过氢氧化钠(碳酸氢钠和/或其他碱性试剂也适用)将pH校正为6.0-7.0。
表1报告了活细胞中的最终浓度和使用上述共混物A、B和C中指示的细菌物种在一步或两步发酵中获得的生长增加。
表1-一步发酵工艺相对于两步发酵工艺中组成共混物的不同共混物(由三种细菌物种组成)的生长比较。在两种测试工艺的发酵结束时进行活细菌的总计数,一步发酵工艺基于食品底物中三种细菌物种的同时接种,而两步发酵工艺基于使用鼠李糖乳杆菌物种的第一发酵,然后是完全灭活该物种,用格氏乳杆菌和短双歧杆菌发酵。
表1中报告的结果表明,两步发酵确保了共混物的整体性能更好,并增加了活细胞中的最终含量。即使获得的发酵食品不含活细胞而仅包含来自微生物发酵的代谢产物,发酵工艺结束时活微生物细胞中的浓度也保证了本发明的发酵食品预期的最终质量。很明显,基于物种的共混物的相同组成,不同的菌株提供了不同的结果,但所有这些都能够确保在两步发酵工艺中比一步发酵结果更好。
表2报告了组成共混物的单一细菌物种的最终浓度,该浓度通过对实验室生长培养基上的活细胞进行选择性计数来测定。同样,与三个物种同时加到底物中的一步工艺相比,两步发酵确保了短双歧杆菌和格氏乳杆菌的生长得到改善(分别为3.32log10 CFU和0.7log10 CFU)。
表2-一步发酵工艺相对于两步发酵工艺中组成共混物的三种细菌物种的生长比较(作为3个不同实验的平均值)。
实施例2-双歧杆菌产生潜力的评价(牛奶中的生长测定)
双歧杆菌产生(bifidogenic)活性是选择性支持双歧杆菌生长的特性,双歧杆菌是一种天然存在于人类肠道中的细菌属,可产生有益代谢物,如短链脂肪酸,对结肠细胞(colonocytes)的存活至关重要,双歧杆菌产生活性通过添加如实施例1获得的发酵粉末到商业牛奶配方(中国和韩国液体配方)进行测试。然后用参考双歧杆菌(菌株短双歧杆菌LMG P-30999和短双歧杆菌LMG S-29966)的洗涤培养物接种改良牛奶,并在24小时内检查和记录它们的生长。将改良牛奶的结果与之前使用相同的普通牛奶配方获得的结果进行比较。测试是针对第一阶段配方牛奶进行的。
表3–双歧杆菌(参考菌株短双歧杆菌LMG S-29966和短双歧杆菌LMG P-30999)在添加最终浓度为1%的粉状发酵成分的配方牛奶第1阶段(适用于0-6个月婴儿)中的生长。
图1显示双歧杆菌在添加1%本发明的发酵成分的第1阶段液体配方牛奶中的生长,表示为log10 CFU,归一化为添加新成分的配方相对于商业配方中微生物共混物的生长差异。
与从市场上获得的配方牛奶(基本配方)相比,发酵成分的存在使双歧杆菌在牛奶中生长得更好。图1显示,与所考虑的商业品牌无关,双歧杆菌在发酵成分的存在下生长得更好。取决于用于发酵的共混物,生长增加(growth gain)为0.31log10 CFU至0.95log10CFU。因此,在所有测试的实验条件下都证实了发酵成分支持双歧杆菌生长的能力(双歧杆菌产生能力)。
实施例3-通过粪便发酵模型(FMF)对双歧杆菌产生潜力的评价
还通过FMF粪便发酵检查了本发明的发酵粉末的双歧杆菌产生能力。采用肠道发酵的实验室模型来研究摄入发酵燕麦成分如何通过促进有益细菌来影响肠道微生物群的组成,从而对抗病原菌或潜在有害微生物。
在新鲜生长培养基存在的情况下,以1%的最终浓度添加如实施例1中获得的成分至标准化混合粪便样本,模拟消费者假定的食物。在适当条件下温育后(在37℃下厌氧生活24小时),通过定量PCR监测主要菌群的生长,比较它们在零时间时和在发酵粉存在下温育后的浓度。因此,评价主要菌群的波动,以评估新成分对人类肠道微生物群的潜在有益和/或有害影响。
厚壁菌门/拟杆菌门(F/B)的比率代表了该成分对人类肠道微生物群影响的评估的第一线。该比率的变化与严重的失调病症例如与肥胖症、肠道慢性疾病等相关的病症有关。正常值约为10,而显著变化的比率(例如,高于100的值)表明厚壁菌门异常盛行,通常与一些疾病相关。
在FMF模型中检查实施例1的发酵食品以排除其对F/B比率的影响。成分的安全性还通过以下事实得到证实:F/B比率在测定过程中没有改变,与用于天然底物发酵的共混物的组成无关。在健康人供体的粪便中在测定开始时,以8.0log10 CFU对厚壁菌门进行量化,和在葡萄糖存在下测定后为7.6。发酵成分作为碳源的存在导致值范围为7.4–7.7log10CFU。在7.8log110 CFU中发现在零时间时存在拟杆菌。测定完成后,拟杆菌在葡萄糖存在下为6.5log10,在发酵成分存在下为6.4–6.8log10。
因此,F/B比率在测定开始时计算为平均1.03,在与葡萄糖温育后增加到1.16。发酵成分的存在导致值为1.11-1.17。已确认该比例不会因发酵成分的存在而改变。
还评估了发酵成分对总细菌的影响,以排除对双歧杆菌以外的细菌群产生的潜在有害影响。观察到总细菌略有增加,正如在葡萄糖存在下预期的那样,总细菌增加了0.3log10。发酵成分决定了总细菌生长增加0.3–0.4log10,类似于葡萄糖(参考碳源)所确定的。
如上所述,通过FMF模型中双歧杆菌的直接定量证实了新发酵成分的双歧杆菌产生作用。表4提供了通过实时定量PCR对双歧杆菌进行定量的数据。
表4-通过比较作为常规碳源的葡萄糖和新发酵成分对FMF模型中双歧杆菌进行定量。还测定了时间为零时的浓度以及在非发酵天然成分存在下获得的值。
碳源 | 平均拷贝数 | FD | 每克FMF计数 | log | 生长增加(log10) |
葡萄糖 | 5.05E+06 | 46.9 | 4.74E+08 | 8.7 | 1.1 |
NF粉末 | 1.67E+06 | 46.8 | 1.57E+08 | 8.2 | 0.6 |
共混物A | 2.48E+06 | 44.5 | 2.21E+08 | 8.3 | 0.7 |
共混物B | 3.40E+06 | 46.5 | 3.17E+08 | 8.5 | 0.9 |
共混物C | 3.91E+06 | 57 | 4.45E+08 | 8.6 | 1.0 |
共混物D | 3.55E+06 | 61.4 | 4.36E+08 | 8.6 | 1.0 |
T0 | 3.66E+05 | 53 | 3.88E+07 | 7.6 | \ |
暴露于发酵成分后,双歧杆菌在很大程度上受到刺激。获得了仅略低于葡萄糖的生长增加的类似结果。
还检查了发酵成分对大范围最终浓度的影响,以证明其功效是剂量依赖性的,以及这种关联以何种方式表达。
表5-通过比较发酵成分的不同最终浓度对FMF模型中双歧杆菌进行定量。检查葡萄糖作为常规碳源的阳性对照。还测定了双歧杆菌的时间为零时的水平以及在非发酵成分存在下获得的值(用作阴性对照)。
在0.5%至5%的最终浓度范围内证实发酵成分对双歧杆菌的刺激。在激活双歧杆菌方面的最佳结果是通过3%的最终浓度实现的,这可以被认为是消费者每天假定的最佳剂量,以诱导积极的双歧杆菌产生作用。
实施例4-与相同的天然非发酵底物相比消化率的改善
评价本发明的食品成分的抗营养素含量。植酸是植物种子中肌醇和储存磷的主要来源,占总磷的70%。谷物和植物源性食品中植酸的丰度是食品和动物饲料行业中关心的问题,因为这种形式的磷由于缺乏专用于植酸脱磷酸化的酶,内源性植酸酶而无法用于单胃动物。此外,植酸的强螯合特性降低了其他必需膳食营养素的生物利用度,例如矿物质,例如Ca2+、Zn2+、Mg2+、Mn2+、Fe2+/3+、蛋白质和氨基酸。该测定是通过Megazyme试剂盒进行的,用于测定植酸化合物。
如表5所示,食物底物的抗营养物如植酸的水平在通过选择的共混物发酵后降低。通过比较未发酵的底物与根据本发明的发酵食物来比较不同的植物底物。在以燕麦为基础的发酵成分中检测到最低浓度的植酸,每100克发酵成分的值为1.34和1.17克。同样在水稻中,根据本发明的发酵后植酸水平降低。
表6-参考不同类型的植物,与非发酵天然底物相比,发酵成分中植酸的最终浓度的降低。
正如预期的那样,发现胡萝卜含有低水平的植酸,在发酵后几乎消失了。其他底物例如天然携带大量植酸的谷物实现了显著减少。用选定的共混物发酵底物导致发酵粉末的产生,这些发酵粉末在抗营养素化合物中自然降低。降低是显著的实体,因为植酸被一些测试的微生物共混物减半。
实施例5比较测试
燕麦水性底物已经与根据本发明的共混物与根据EP 1169925的共混物以及根据本发明的共混物发酵。
测试是根据US 2010098805中公开的方法或根据本发明的方法进行的。
根据US 2010098805的两步发酵
将基于燕麦的水性底物在37℃下温育24小时,以便让天然存在的微生物在底物内发育。温育后,自然发酵的生物质经过巴氏杀菌,以灭活活细菌。然后用同时混合在一起的菌株干酪乳杆菌(L.casei)ATCC334、嗜酸乳杆菌(L.acidophilus)ATCC4356、短双歧杆菌(B.breve)ATCC15700、长双歧杆菌(B.longum)ATCC15707、长双歧杆菌婴儿亚种(B.longumsub.infantis)ATCC15697(共混物B)接种第一次发酵所得的产物(第二发酵步骤)。对不同的选择性培养基进行活菌计数以精确量化接种物的大小。MRS琼脂用于总乳酸杆菌属(Lactobacillus spp.)的活菌计数。补充有10mg/ml万古霉素的MRS用于计数干酪乳杆菌ATCC334,补充有0.1和10μg/ml克林霉素和环丙沙星的MRS用于计数嗜酸乳杆菌ATCC4256。补充有50mg/l莫匹罗星的TOS丙酸盐琼脂基础培养基(Sigma-Aldrich Merck)用于双歧杆菌属的计数。
在微需氧条件下,将发酵块(fermentation bulk)在37℃下温育24小时。温育后,对上述选择性培养基进行活菌计数以评价微生物达到的最终浓度。然后如先前所述灭活发酵的生物质。在第二发酵中使用由格氏乳杆菌L6、鼠李糖乳杆菌L13b和短双歧杆菌2TA组成的共混物(共混物A)重复相同的步骤。
根据本发明的两步发酵
使用共混物A重复实施例1的工艺。共混物B也进行类似的工艺,在第一发酵步骤中使用副干酪乳杆菌(L.paracasei)ATCC334代替鼠李糖乳杆菌。事实上,副干酪乳杆菌与鼠李糖乳杆菌具有最接近的系统发育相似性。
结果
通过本发明和US 2010098805的方法的两种共混物的生长(以log10 CFU计)在表1中报告。数据是通过测试中各菌株的生长时间T24和时间T0(发酵开始)之间的平板小数计数差异获得的。已通过定量PCR测定了共混物B的三种双歧杆菌菌株。
表1中的数据表明,US 2010098805的发酵方法不允许共混物A的3个菌株中的2个生长。
相反,本发明的两步发酵方法能够使EP 1169925中公开的共混物A的三种菌株以及共混物B的菌株的生长与物种鼠李糖乳杆菌(系统发育类似于副干酪乳杆菌)和格氏乳杆菌(系统发育类似于嗜酸乳杆菌)的共混物A的生长具有相同的程度。
此外,与共混物B中包含的其他双歧杆菌相比,本发明的方法能够更有效地发酵短双歧杆菌。
当采用相同的发酵方法和条件时,共混物B无法复制共混物A的性能。
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Claims (12)
1.一种发酵食品或食品补充剂,其包含通过以下方式获得的植物底物:用鼠李糖乳杆菌(L.rhamnosus)进行第一发酵步骤,然后用由格氏乳杆菌(Lactobacillus gasseri)和短双歧杆菌(Bifidobacterium breve)组成的共混物进行第二发酵步骤以及最终灭活细菌细胞。
2.根据权利要求1所述的发酵食品或食品补充剂,其中所述植物底物选自燕麦、大麦、玉米、谷物、假谷物、无麸质蔬菜、蔬菜、水果、坚果。
3.根据权利要求1所述的发酵食品或食品补充剂,其中所述植物底物是燕麦。
4.根据权利要求1所述的发酵食品或食品补充剂,其中所述植物底物是与选自玉米油、葵花籽油或任何其他植物油的油组合的燕麦。
5.根据权利要求1至4中任一项所述的发酵食品,呈液体、粉末或奶油形式。
6.根据权利要求1至4中任一项所述的发酵食品,其用作食品成分和/或食品补充剂,终浓度为0.5至20%。
7.根据权利要求1至6中任一项所述的发酵食品或食品补充剂,其中益生菌共混物中格氏乳杆菌、鼠李糖乳杆菌和短双歧杆菌的CFU比为1:1:1。
8.根据权利要求1至5中任一项所述的发酵食品或食品补充剂,其中所述益生菌共混物由加氏乳杆菌、鼠李糖乳杆菌和短双歧杆菌组成。
9.根据权利要求1至6中任一项所述的发酵食品或食品补充剂,其为饮料、奶油、涂抹酱、粉末形式。
10.根据权利要求1至7中任一项所述的发酵食品或食品补充剂,还含有驴奶或羊奶。
11.根据权利要求1至7中任一项所述的发酵食品或食品补充剂,其用于改善自然母性、哺乳和婴儿断奶。
12.一种制备根据权利要求1至7所述的发酵食品或食品补充剂的方法,包括用鼠李糖乳杆菌发酵植物底物,然后用格氏乳杆菌和短双歧杆菌进行第二发酵步骤,然后灭活细菌细胞。
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WO2020201284A1 (en) | 2020-10-08 |
CN113710102B (zh) | 2024-05-07 |
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