WO2008116356A1 - Procédé de préparation d'un lait fermenté stable à température ambiante présentant des taux élevés de bactéries d'acide lactique viables - Google Patents

Procédé de préparation d'un lait fermenté stable à température ambiante présentant des taux élevés de bactéries d'acide lactique viables Download PDF

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WO2008116356A1
WO2008116356A1 PCT/CN2007/002037 CN2007002037W WO2008116356A1 WO 2008116356 A1 WO2008116356 A1 WO 2008116356A1 CN 2007002037 W CN2007002037 W CN 2007002037W WO 2008116356 A1 WO2008116356 A1 WO 2008116356A1
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Prior art keywords
milk
added
lactobacillus rhamnosus
concentration
fermented
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PCT/CN2007/002037
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English (en)
French (fr)
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Haibo Ling
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Beijing Ferment Bio-Tech.Com, Ltd
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Publication of WO2008116356A1 publication Critical patent/WO2008116356A1/zh
Priority to US12/564,762 priority Critical patent/US10072310B2/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/175—Rhamnosus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the present invention relates to the field of fermented milk, and in particular to a method for preparing a fermented milk beverage which maintains a high viable cell count at normal temperature. Background technique
  • Fermented milk is a popular fermented dairy product that is popular among consumers.
  • the popularity of such products is largely related to their active lactic acid bacteria that are beneficial to the human body. It is generally accepted that at least 10 5 -10 7 cfli of active lactic acid bacteria should be contained per liter (or per gram) of product, and at least 100 liters (or grams) of product should be consumed per day in order to be effective for human health. Therefore, it is very important to ensure the survival of these lactic acid bacteria during transportation and storage.
  • Some methods have been reported to increase the number of lactic acid bacteria. These methods include: adding an oxygen scavenger (such as Vc), adding prebiotics (such as oligosaccharides), encapsulating lactic acid bacteria into microcapsules, without adding Lactobacillus bulgaricus and reducing Oxidation-reduction potential (eg, cysteine) (NP Shah, Probiotic bacteria: Selective enumeration and survival in dairy foods, J. Dairy Sci” 83:894-907, 2000; A. Lourens and BC Viljoen, Int. Dairy J., 11 : 1-17, 2001 ). However, these methods have had little effect.
  • Vc oxygen scavenger
  • prebiotics such as oligosaccharides
  • lactic acid bacteria can be fermented by lactose, and lactose fermentation produces lactic acid, which causes the pH of the milk to drop rapidly, which directly leads to a decrease in the number of lactic acid bacteria that survive.
  • lactose fermentation produces lactic acid, which causes the pH of the milk to drop rapidly, which directly leads to a decrease in the number of lactic acid bacteria that survive.
  • Streptococcus thermophilus and/or Lactobacillus bulgaricus as a fermenting agent
  • fermenting milk at around 43 °C will produce a large amount of lactic acid in 3-8 hours, causing the pH of the milk to rapidly drop below 4.5, leading to lactic acid bacteria. Decreased activity or even death. .
  • controlling the acid production of fermented milk is the key to solving the problem of fermented milk stored at room temperature and maintaining a high number of viable bacteria.
  • Lactobacillus rhamnosus (available from the American Type Culture Collection, strain number ATCC 53 103) is a probiotic isolated from healthy humans (US patent 4,839,281; US patent 5,032,399) and can be removed Endotoxin in plasma (US patent 5, 413, 785).
  • the biggest feature of this bacterium is that it does not ferment lactose, and it is resistant to acid and oxygen.
  • the optimum growth temperature for this strain is 37 degrees Celsius and the growth rate is fast. Because this bacterium has the function of preventing and treating gastrointestinal discomfort (such as diarrhea) and improving immunity, it is used in health care products and functional foods (such as yogurt).
  • gastrointestinal discomfort such as diarrhea
  • functional foods such as yogurt
  • An object of the present invention is to provide a method for preparing a fermented milk beverage which maintains a high viable cell count at a normal temperature.
  • Post-acidification After the fermentation process of yoghurt or fermented milk is stopped, the yogurt or milk yoghurt in the product continues to produce acid, which leads to acidification of the product. It is also called acidification after production or acidification after fermentation.
  • normal temperature means a temperature which can be attained in a room under normal conditions, for example, 4 to 40 ° C, usually 10 to 30 ° C, and typically 12 to 25 ° C.
  • the principle of the present invention is: first use ordinary yogurt bacteria [by Lactobacillus bulgaricus (ctob c/// ⁇ Bulgaric s) and/or a single or mixed species of Streptococcus thermophilus] fermented, diluted, formulated and sterilized, and added to Lactobacillus rhamnosus ⁇ Lactobacillus rl ninosus" under aseptic conditions ATCC 53103, which utilizes the monosaccharide produced by the fermentation of common yoghurt bacteria to proliferate and produce lactic acid, so that the product acidity is finally stabilized in the product design acidity.
  • the method of the invention firstly adds the common yoghurt bacteria to the milk sample for fermentation, fermenting to a pH of 3.8-4.8, adding a part of the sweetener, the essence and the thickening emulsion stabilizer for dilution and preparation, and then sterilizing.
  • Concentrated liquid cultures, concentrated frozen cultures or freeze-dried bacterial powders containing ATCC 53103 (purchased from the American Type Culture Collection) containing Lactobacillus rhamnosus Rantobacillus rhanmosus (acquired from the American Type Culture Collection) were added to the formulated milk under aseptic conditions.
  • the PH value during the preparation is adjusted according to the pH drop value which can be caused by the amount of the monosaccharide contained in the fermentation liquid, for example, the amount of monosaccharide contained in the fermentation liquid may cause the blending.
  • the pH value of the milk is decreased by 0.2, and the pH value of the prepared milk product is set to 4.2.
  • the pH value is set at 4.4 when the preparation is performed, because Lactobacillus rhanmosus ATCC 53103 gradually produces acid in the formulated milk.
  • the pH is stabilized at the finished product set value until the monosaccharide in the blended milk is consumed.
  • the milk sample is fresh milk or reduced milk (including but not limited to milk or goat milk or reduced milk) or concentrated whey protein is added to fresh milk or reduced milk, and the concentration of protein in the milk sample is 0.1. -7%, milk solids concentration of 1 -30%.
  • the concentration of protein in the milk sample is from 0.7 to 4.5% and the concentration of milk solids is from 6 to 20%.
  • the common yogurt bacteria are any bacteria known to those skilled in the art for producing yogurt, preferably Lactobacillus bulgaricus, and/or Streptococcus thermophihis.
  • the amount of force is 10 1 - 10 9 cfu / ml milk sample.
  • the fermentation temperature of common yogurt bacteria is 30-45 ° C, and the fermentation time is 2.5-10 hours.
  • the dilution and blending are based on fermented milk, adding a part of sweetener, sour agent, essence, thickening emulsion stabilizer and water, and the protein concentration is adjusted to 0.7-4.5%, and the fermented milk is diluted 1.5-20 times.
  • the amount of addition of ATCC 53103 of Lactobacillus rhamnosus actobaciUus rhanmosus is not subject to any theory, and according to the principle of cost-effectiveness, the addition amount of Lactobacillus rhanmosus) ATCC 53103 is preferred.
  • Addition of ATCC 53103 The amount is 10 5 - 10 7 cfu/ml milk sample.
  • the acid is gradually produced until the monosaccharide in the blended milk is consumed, and the pH varies between 0.1 and 1.0.
  • the fermented milk beverage prepared according to the method of the present invention is stored at room temperature (10-30 ° C) for 1 to 6 months, and the viable cell count is at least 10 5 C f ml milk sample, and the storage time and the viable count index are high.
  • the fermented milk beverage obtained by the conventional method is effectively solved the problem that the fermented milk beverage has a short shelf life and a low number of viable bacteria at normal temperature.
  • Example 1 Fermented milk beverage prepared by adding Lactobacillus rhamnosus after fermentation sterilization of common yogurt bacteria and its storage experiment
  • the lyophilized powder of Lactobacillus rhamnosus actobacillus rhamnosus ATCC 53103 was inoculated into the above emulsion according to a 5 10 5 cfu/ml milk sample. Inspect at 12 ⁇ 25°C The pH value and the number of viable cells were measured, and the results are shown in Table 1. The results showed that after storage for 8 weeks at 12 to 25 ° C, the viable count of Lactobacillus rhamnosus was at least 10 0 C f ml, and the pH of the product was above 3.7.
  • Example 2 Fermented milk beverage prepared by adding Lactobacillus rhamnosus after fermentation sterilization of common yogurt bacteria and its storage experiment
  • Example 2 In the same manner as in Example 1, the whole milk powder concentration was 25%, and the whole milk was reduced.
  • the common yoghurt BF-306 was inoculated (purchased from Beijing Vermont Biotechnology Co., Ltd., by Streptococcus thermophilus and Lactobacillus bulgaricus, which is fermented to a pH of 3.8.
  • the reduced fermented milk was diluted 9 times to prepare a milk beverage of protein 0.7, and the pH of the mixture was adjusted to 4.4 with 10% citric acid, homogenized and sterilized according to the method of Example 1, and the Lactobacillus rhamnosus was prepared.
  • Lactobacillus rhamnosus A concentrated culture of ATCC 53 103 was inoculated into the above emulsion according to a latex sample of 1.5 x 10 7 cfu/ml. The pH value and the viable cell count were examined at 12 to 25 ° C, and the results are shown in Table 2. The results showed that the viable count of Lactobacillus rhamnosus was at least 10 7 cfo/ml after storage for 6 months at 12 ⁇ 25 Q C. The pH is above 4.2. Table 2 Survival of lactic acid bacteria and product pH detection data
  • Example 3 Fermented milk beverage prepared by adding Lactobacillus rhamnosus after fermentation sterilization of common yogurt bacteria and its storage experiment
  • Example 2 In the same manner as in Example 1, the milk solids were added to 1.5% of fresh milk, 3.5% of whole milk powder, and 15% of whole milk, and inoculated with common yoghurt BF-301 (purchased from Beijing Vermont Bio) Technology Co., Ltd., by Streptococcus thermophilus 5"ire iococc”s And Lactobacillus bulgaricus 3 ⁇ 4 j3 ⁇ 4 ), fermented to a pH of 4.0.
  • common yoghurt BF-301 purchased from Beijing Vermont Bio
  • the reduced fermented milk was diluted 5.5 times, and the milk beverage was formulated into protein 0.7, and the pH of the mixture was adjusted to 4.3 with 10% citric acid.
  • the rhamnosose milk was prepared.
  • Bacillus actobacillus rhamnosiis) ATCC 53103 was inoculated into the above emulsion according to a 2 x 10 6 cfu/ml milk sample.
  • the pH value and viable cell count were measured at 37 ° C for 15 days, and the results are shown in Table 3. The results showed that after 15 days of storage at 37 ° C, the viable count of Lactobacillus rhamnosus was at least 10 7 cfii / ml milk sample, and the pH value of the product was above 3.9.
  • Example 2 In the same manner as in Example 1, the whole milk powder concentration was 28%, and the whole milk was reduced.
  • the yoghurt BF-208 (purchased from Beijing Vermont Biotechnology Co., Ltd., consisting of Streptococcus thermophilus) was fermented. To pH 4.5.
  • the reduced fermented milk was diluted 4 times according to the standard of 1.8% of the finished protein, and after blending and sterilizing, Lactobacillus rhamnosus actobaciUus rhamnosus) ATCC 53103 was inoculated into the above emulsion according to a 5 x 10 6 cfu/ml milk sample.
  • the pH value and the viable cell count were measured at 12 to 25 ° C, and the results are shown in Table 4. The results showed that after storage for 6 months at 12 to 25 ° C, the viable count of Lactobacillus rhamnosus was at least 10 7 cfb / ml, and the pH of the product was above 4.3.

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Description

一种在常温下保持高活菌数的发酵乳饮料的制备方法 技术领域
本发明涉及发酵乳领域， 具体地涉及一种在常温下保持高活菌数的发酵 乳饮料的制备方法。 背景技术
发酵乳， 特别是酸奶， 是广受消费者欢迎的发酵乳制品。 此类产品之所 以受欢迎， 很大程度上是与它们含有对人体有益的活性乳酸菌有关。 通常认 为， 每亳升 （或每克）产品中应含有至少 105-107cfli 的活性乳酸菌， 每天应 消费至少 100亳升（或克）产品， 才能对人体健康起效。 因此， 在运输、 贮 藏过程中确保这些乳酸菌的存活非常重要。
然而， 在目前冷链不健全的条件下， 当产品在非冷藏条件下贮藏较长时 间后，发酵乳中的乳酸菌的存活率常常很低，甚至检测不到活菌（ W.T. Hamann 和 E.H. Marth, Survival of Streptococcus thermophilus and Lactobacillus bulgaricus in commercial and experimental yoghurts, J. Food Protection, 47:781-786, 1984 )。产生这一现象1的原因是非冷链条件贮藏使得酸奶产品后酸 化严重， 而过度酸化会加速乳酸菌的死亡。 这一点已在研究报告中得到了充 分说明（郭清泉和张兰威， 引起酸奶制品发生后酸化的主要发酵剂菌及性质， 食品与机械， 87 ( 1 ): 14-16 , 2002; W.T. Hamann 和 E.H. Marth, J. Food Protection, 47:781-786, 1984 )„ 许多发展中国家缺乏完善的冷链系统， 因此发 酵奶在流通过程中常常处于非冷藏状态， 导致产品质量下降， 产品的货架期 大大缩短。
已有一些文献报道了增加乳酸菌存活数量的方法， 这些方法包括： 添加 除氧剂 （如 Vc ), 添加益生元 （如寡糖类）， 将乳酸菌包入微胶囊中， 不加保 加利亚乳酸杆菌和降低氧化还原电位（如加半胱氨酸）等（ N.P. Shah, Probiotic bacteria: Selective enumeration and survival in dairy foods, J. Dairy Sci" 83 :894-907, 2000; A. Lourens 和 B.C. Viljoen, Int. Dairy J., 11 : 1-17, 2001 )。 然 而， 这些方法收效甚微。
问题的根源在于大部分的乳酸菌都能利用乳糖发酵， 而乳糖发酵会产生 乳酸， 从而使得牛乳的 pH值迅速下降， 直接导致存活的乳酸菌数量下降。 例如釆用嗜热链球菌和 /或保加利亚乳杆菌作发酵菌剂， 在 43°C左右发酵牛 奶， 会在 3 - 8小时内产生大量的乳酸， 使得牛奶 pH值迅速下降到 4.5以下， 导致乳酸菌活性下降甚至死亡。 .
因此， 控制发酵乳产酸成为解决发酵乳常温贮藏并保持高活菌数问题的 关键。
鼠李糖乳杆菌（ Lactobacillus rhamnosus ) (可购自美国菌种保藏中心， 菌 株编号为 ATCC 53 103)是一株从健康人体分离出来的益生菌 （US patent 4,839,281 ; US patent 5,032,399), 并且可以去除血浆里的内毒素 （US patent 5， 413 , 785 ) 。 此菌的最大特点是不发酵乳糖， 而且耐酸和耐氧。 此菌的最佳 生长温度为 37摄氏度， 生长速度快。 因为此菌具有预防和治疗肠胃不适（如 腹泻） 以及提高免疫力的功能， 故此菌被用于保健品和功能食品 （如酸奶） 中。 关于此菌的详细的生物学特性和益生功能， 请参考上述专利文书和综述 文章 - M. Saxelin, Lactobacillus GG A human probiotic strain with thorough clinical documentation, Food Rev. Int., 13 :293-3 13, 1997. 发明内容
(一）要解决的技术问题
本发明的目的是提供一种在常温下保持高活菌数的发酵乳饮料的制备方 法。
(二）技术方案
在本发明的意义上， 对所涉及的技术术语作如下定义：
后酸化： 指酸奶或发酵乳生产发酵过程停止之后， 产品内的酸奶菌或乳 酸奶继续产酸而导致产品酸化， 又称生产后酸化或发酵后酸化。
在本发明中， 术语 "常温" 是指正常情况下室内所能够达到的温度， 例 如 4〜40 °C， 通常为 10〜30 °C , 典型的是 12〜25°C。
本发明的原理是：首先利用普通酸奶菌 [由保加利亚乳杆菌（ ctob c///^ bulgaric s )和 /或嗜热链球菌 Streptococcus thermophilus 的单一菌种或混合 菌种组成]进行发酵， 经稀释、 调配并灭菌后， 在无菌条件下加入鼠李糖乳杆 菌 {Lactobacillus rl ninosus") ATCC 53103, 使之利用普通酸奶菌发酵产生 的单糖来进行增殖， 并产生乳酸， 使产品酸度最终稳定在产品设计酸度。
本发明所述的方法， 是先向乳样中加入普通酸奶菌进行发酵， 发酵到 PH 值 3.8-4.8终止， 添加部分甜味剂、 香精和增稠乳化稳定剂进行稀释和调配 后灭菌， 在无菌条件下向调配乳中加入含有鼠李糖乳杆菌 Lactobacillus rhanmosus) ATCC 53103 (购自美国菌种保藏中心） 的浓缩液态培养物、 浓缩 冷冻培养物或冷冻干燥菌粉。
本发明所述的方法，所述调配时 PH值会根据发酵液中含有的单糖量所能 导致的 PH下降值设定调配时的 pH值， 例如发酵液中所含单糖量会导致调配 乳 pH值下降 0.2， 调配乳成品的 pH值欲设定在 4.2, 则调配时 pH值设定在 4.4， 因为鼠李糖乳杆菌 ( Lactobacillus rhanmosus) ATCC 53103的在调配 乳中会逐渐产酸， 直至调配乳中的单糖被消耗后 PH值稳定在成品设定值。
其中， 所述乳样为鲜奶或还原奶 （包括但不限于牛乳、 羊乳的鲜奶或还 原奶） 或在鲜奶或还原奶中添加浓缩乳清蛋白， 乳样中蛋白质的浓度为 0.1-7%, 乳固体浓度为 1 -30%。 优选地， 乳样中蛋白质的浓度为 0.7-4.5%， 乳固体浓度为 6-20%。
在本发明所述的方法中， 所述普通酸奶菌为本领域技术人员已知的任何 用于生产酸奶的菌， 优选保加利亚乳杆菌 Lactobacillus bulgaricus、和 /或嗜 热链球菌 ( Streptococcus thermophihis ), 其添力口量均是 101— 109cfu/ml乳样。
普通酸奶菌的发酵温度为 30-45°C, 发酵时间为 2.5-10小时。
所述稀释和调配是以发酵乳为基料， 添加部分甜味剂、 酸味剂、 香精、 增稠乳化稳定剂和水， 调配至蛋白质浓度为 0.7-4.5%， 发酵乳稀释 1.5-20倍。
在本发明所述的方法中， 鼠李糖乳杆菌 actobaciUus rhanmosus ) ATCC 53103 的添加量并不受到任何理论的限制， 根据经济有效的原则， 优选鼠李 糖乳杆菌 Lactobacillus rhanmosus ) ATCC 53103的添加量是 101 - 109cfu/ml 乳样， 更优选， 鼠李糖乳杆菌
Figure imgf000004_0001
r½7W70« ) ATCC 53103的添加 量是 105 - 107cfu/ml乳样。 逐渐产酸， 直至调配乳中的单糖被消耗掉， pH值变化范围在 0.1〜1.0之间。
(三）有益效果
根据本发明所述的方法制备的发酵乳饮料， 在室温下 （ 10_30°C) 贮藏 1〜6个月， 活菌数至少为 105 Cf ml乳样， 储藏时间和活菌数指标均高于釆用 常规方法发酵得到的发酵乳饮料， 有效解决了发酵乳饮料在常温下货架期短 且活菌数低的难题。 具体实施方式
以下实施例用于说明本发明， 但不用来限制本发明的范围。 实施例 1 普通酸奶菌发酵灭菌后再添加鼠李糖乳杆菌制备的发酵乳饮料 及其存放实验
将 8%的全脂奶粉和 3%的浓缩乳清蛋白 （WPC-80, 蛋白含量 80%)加 入水中，配成还原全脂牛奶，取 1000 ml还原全脂牛奶在 115°C加热 15分钟， 冷却后， 接种普通酸奶菌 BF-203 (购自北京弗蒙特生物技术有限公司， 由嗜 热链球菌 Streptococcus ίΛ »σρ?ϋ"·? 禾口保力口利亚乳杆菌 Lactobacillus bulgaricus ,发酵至 PH值 4.7, 冷却至 25°C以下。
用 400ml 70°C的水溶解酸性乳饮料稳定剂 SY-806 (购自北京弗蒙特生 物技术有限公司）6g， AK糖（购自张家港浩波化学品公司）0.5g,搅拌 20min， 冷却至 25°C以下， 水合 30min后， 按照成品蛋白浓度为 1.0%的标准 （全脂 奶粉蛋白含量按 26%计算）， 将上述还原发酵乳稀释 2.5倍， 配成蛋白 1.0的 乳饮料， 并加入上述稳定剂溶液中， 用 10%柠檬酸将混合液 pH值调至 4.4， 用水定容至 1000ml, 搅拌 5min。 预热至 70°C, 进行均质， 均质压力 20MPa。 然后灭菌， 条件为 115°C、 15min。
再冷却至 37°C以下， 将鼠李糖乳杆菌 actobacillus rhamnosus) ATCC 53103冻干菌粉按照 5 105cfu/ml乳样接种至上述乳液中。 在 12〜25°C考察 测定 pH值和活菌数， 结果见表 1。 结果表明， 在 12〜25°C贮藏 8周后， 鼠李 糖乳杆菌的活菌数至少为 100 Cf ml乳样， 产品 pH值在 3.7以上。
表 1 乳酸菌的存活情况和产品 pH值检测数据
Figure imgf000006_0001
实施例 2 普通酸奶菌发酵灭菌后再添加鼠李糖乳杆菌制备的发酵乳饮料 及其存放实验
与实施例 1方法相同， 全脂奶粉浓度为 25%配成还原全脂牛奶， 接种普 通酸奶菌 BF-306 (购自北京弗蒙特生物技术有限公司， 由嗜热链球菌 Streptococcus thermophilus 禾口保力口禾 lj亚乳杆菌 Lac tobacillus bulgaricus 构 成）， 发酵至 PH值 3.8。
将上述还原发酵乳稀释 9倍， 配成蛋白 0.7的乳饮料， 用 10%柠檬酸将 混合液 pH值调至 4.4, 按实施例 1 的方法均质并灭菌后， 将鼠李糖乳杆菌 ( Lactobacillus rhamnosus ) ATCC 53 103 的浓缩培养物按照 1. 5 x 107cfu/ml 乳样接种至上述乳液中。 在 12〜25°C考察测定 pH值和活菌数， 结果见表 2。 结果表明，在 12〜25QC贮藏 6个月后，鼠李糖乳杆菌的活菌数至少为 107cfo/ml pH值在 4.2以上。 表 2 乳酸菌的存活情况和产品 pH值检测数据
Figure imgf000007_0003
实施例 3 普通酸奶菌发酵灭菌后再添加鼠李糖乳杆菌制备的发酵乳饮料 及其存放实验
与实施例 1方法相同， 乳固体为 1 1.5%的鲜奶中再加入 3.5%的全脂奶粉 配成浓度为 15%的全脂牛奶， 接种普通酸奶菌 BF-301 (购自北京弗蒙特生物 技术有限公司， 由嗜热链球菌 5"ire iococc"s
Figure imgf000007_0001
和保加利亚乳杆菌 Lactobacillus bulgaricus ¾ j¾ ) , 发酵至 pH值 4.0。
将上述还原发酵乳稀释 5.5倍， 配成蛋白 0.7的乳饮料， 用 10%柠檬酸将 混合液 pH值调至 4.3， 按实施例 1的方法均质调配并灭菌后， 将鼠李糖乳杆 菌 actobacillus rhamnosiis ) ATCC 53103按照 2 x 106cfu/ml乳样接种至上述 乳液中。 在 37°C考察 15天测定 pH值和活菌数， 结果见表 3。 结果表明， 在 37°C贮藏 15天后， 鼠李糖乳杆菌的活菌数至少为 107cfii/ml乳样， 产品 pH 值在 3.9以上。
表 3 乳酸菌的存活情况和产品 pH值检测数据
考察时间 （天） pH值 活菌数 （ l
Figure imgf000007_0002
乳样）
0 4.3 1 0.15
1 4.17 26 4 3.94 17
7 3.98 21
10 3.93 3
15 3.93 1.1 实施例 4 普通酸奶菌发酵灭菌后再添加鼠李糖乳杆菌制备的发酵乳饮料及 其存放实验
与实施例 1方法相同， 全脂奶粉浓度为 28%配成还原全脂牛奶， 接种普 通酸奶菌 BF-208 (购自北京弗蒙特生物技术有限公司， 由嗜热链球菌 Streptococcus thermophilus构成)， 发酵至 pH值 4.5。
按照成品蛋白 1.8%的标准，将上述还原发酵乳稀释 4倍，调配并灭菌后， 将鼠李糖乳杆菌 actobaciUus rhamnosus ) ATCC 53103 按照 5 x 106cfu/ml 乳样接种至上述乳液中。 在 12〜25°C考察测定 pH值和活菌数， 结果见表 4。 结果表明，在 12〜25°C贮藏 6个月后，鼠李糖乳杆菌的活菌数至少为 107cfb/ml 乳样， 产品 pH值在 4.3以上。
表 4 乳酸菌的存活情况和产品 pH值检测数据
考察时间 （月） pH值 活菌数（107cfb/ml乳样）
0 4.8 0.5
1 4.43 27
2 4.35 29
3 4.40 32
4 4.37 22
5 4.41 15
6 4.33 7.6

Claims

1、 一种在常温下保持高活菌数的发酵乳饮料的制备方法， 其特征在于先 向乳样中加入普通酸奶菌进行发酵， 发酵到 pH值 3.8-4.8终止， 进行稀释和 调配后灭菌， 在无菌条件下向调配乳中加入鼠李糖乳杆菌 actobacilhis rhamnosus ) ATCC 53103。
权
2、 根据权利要求 1所述的方法， 其特征在于所述乳样为鲜奶、 还原奶或 在鲜奶或还原奶中添加浓缩乳清蛋白， 乳样中蛋白质的浓度为 0.1-7%， 乳固 体浓度为 1 - 30%。
3、 根据权利要求 2 所述的方法， 其特征在于乳样中蛋白质的浓度为 0.7-4.5%, 乳固体浓度为 6-20%。
4、 根据权利要求 1所述的方法， 其特征在于所述普通酸奶菌为保加利亚 杆菌 ( Lactobacillus bulgaricus ) 和 /或嗜热链球菌 ( Streptococcus thermophilus )， 其添加量均是 101 - 109cfu/ml乳样。 书
5、 根据权利要求 1 所述的方法， 其特征在于普通酸奶菌的发酵温度为 30-45°C, 发酵时间为 2.5-10小时。
6、 根据权利要求 1所述的方法， 其特征在于所述稀释和调配是将发酵乳 调配至蛋白质浓度为 0.7-4.5%, 发酵乳稀释 1.5-20倍。
7、 根据权利要求 1所述的方法， 其特征在于所述调配乳的成品 pH值最 终控制范围为 3.5-4.7。
8、 根据权利要求 1 所述的方法， 其特征在于所述鼠李糖乳杆菌 ( Lactobacillus rhamnosus ) ATCC 53103的添加量是 101 - 109ci ml乳样。
9、 根据权利要求 8 所述的方法， 其特征在于所述鼠李糖乳杆菌 ( Lactobacillus rhamnosus ) ATCC 53103的添加量是 105 - 107cfu/ml乳样。
10、 根据权利要求 1 -9中任一项所述的方法， 其中所述的鼠李糖乳杆菌 ( Lactobacillus rhamnosus ) ATCC 53103 可以是以含有鼠李糖乳杆菌 ( Lactobacillus rhamnosus ) ATCC 53103的浓缩液态培养物、 浓缩冷冻培养物 或冷冻干燥菌粉的形式被加入。
11、 根据权利要求 1 - 10 中任一项所述方法所制备的在常温下保持高活 菌数的发酵乳饮料。
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US20100009034A1 (en) 2010-01-14
US20100015285A1 (en) 2010-01-21
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US10072310B2 (en) 2018-09-11
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