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含LR1的发酵奶制品生物技术的开发及其在实验中的功能特性评估

[Development of biotechnology of fermented milk product with LR1 and the evaluation of its functional property in experiment and ].

作者信息

Semenikhina V F, Rozhkova I V, Begunova A V, Fedorova T V, Shirshova T I

机构信息

All-Russian Dairy Research Institute, Moscow.

A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow.

出版信息

Vopr Pitan. 2018;87(5):52-62. doi: 10.24411/0042-8833-2018-10053. Epub 2018 Sep 13.

Abstract

In this article the biotechnology of the dairy product based on the probiotic strain of Lactobacillus reuteri LR1 is presented. The following conditions of milk fermentation were screened: fermentation by monoculture of Lactobacillus reuteri LR1, fermentation by monoculture of Lactobacillus reuteri LR1 with addition of yeast extract as growth-promoting factor, and combined fermentation by Lactobacillus reuteri LR1, Lactobacillus helveticus NK1 and Streptococcus thermophilus (HTC). It had been demonstrated that after 8 hours of cultivation the number of Lactobacillus reuteri LR1 cells in the monoculture with introduced yeast extract was up to 5,9×10 CFU/cm whereas cell count for the monoculture without yeast extract introduction was 1,6×10 CFU/cm. The optimized biotechnological parameters of the dairy product fermentation, which provided the required Lactobacillus reuteri LR1 cell count, were as follows: Lactobacillus reuteri LR1 starter dosage of 6%, HTC starter dosage of 3-4%, fermentation temperature of 37±1 °C, and fermentation duration of 6 hours. The developed product possessed an apparent antagonistic activity against test-cultures of such pathogenic and opportunistic microorganisms as E. coli ATCC 25922, Salmonella typhimurium, Staphylococcus aureus ATCC 6538 and Klebsiella pneumoniae. The survival of the test-cultures cultivated with the obtained dairy product on the first cultivation day was from 36 to 46% and on the second - from 8 to 20%, in comparison with the pure test-cultures. The investigation of the functional properties of the obtained dairy product was carried out by the single-factor experiment with the albino Wistar rat-stock (initial body weight 160±10 g, n=10 in each group). Its positive effect on the rats' microbiome composition and lipid exchange indices has been demonstrated. It had been shown that the administration of the obtained dairy product in the rats' diet (5 ml per day per os) during 30 days didn't cause any abnormalities in the health status and behavior of the laboratory animals of spf-category. The number and distribution of leucocytes and lymphocytes, and granulocytes in all rats' populations (intact, control and treatment) were within the normal range. Upon the introduction of the developed dairy product into the rats' diet, the increased levels of bifidobacteria, lactic acid bacteria and typical for normal rats' microflora enterobacteria were observed in the rats' microbiome. As for the biochemical indices that characterize rats' lipid metabolism, the rats consuming fermented dairy products (control and treatment) demonstrated statistically significant reduction of blood serum cholesterol level compared to the intact rat group; additionally the rats consuming the developed in this study dairy product (treatment) demonstrated statistically significant reduction of blood serum triglyceride level compared to the intact rats. Utilized in this study Lactobacillus reuteri LR1 is the first strain that was purified in Russia and characterized as useful for manufacturing of the probiotic food products, since currently only Lactobacillus reuteri strains of foreign origin can be seen on the market.

摘要

本文介绍了基于罗伊氏乳杆菌LR1益生菌株的乳制品生物技术。筛选了以下牛奶发酵条件:罗伊氏乳杆菌LR1单一培养物发酵、添加酵母提取物作为生长促进因子的罗伊氏乳杆菌LR1单一培养物发酵,以及罗伊氏乳杆菌LR1、瑞士乳杆菌NK1和嗜热链球菌(HTC)联合发酵。结果表明,培养8小时后,添加酵母提取物的单一培养物中罗伊氏乳杆菌LR1细胞数高达5.9×10 CFU/cm,而未添加酵母提取物的单一培养物细胞数为1.6×10 CFU/cm。提供所需罗伊氏乳杆菌LR1细胞数的乳制品发酵优化生物技术参数如下:罗伊氏乳杆菌LR1发酵剂用量6%,HTC发酵剂用量3 - 4%,发酵温度37±1℃,发酵时间6小时。所开发的产品对大肠杆菌ATCC 25922、鼠伤寒沙门氏菌、金黄色葡萄球菌ATCC 6538和肺炎克雷伯菌等致病性和机会性微生物的测试培养物具有明显的拮抗活性。与纯测试培养物相比,用所得乳制品培养的测试培养物在第一天培养时的存活率为36%至46% , 第二天为8%至20%。用白化Wistar大鼠种群(初始体重160±10 g,每组n = 10)进行单因素实验,对所得乳制品的功能特性进行了研究。已证明其对大鼠微生物群组成和脂质交换指标有积极影响。结果表明,在大鼠饮食中每天经口给予所得乳制品(5 ml)30天,未导致spf级实验动物的健康状况和行为出现任何异常。所有大鼠群体(完整组、对照组和处理组)中白细胞、淋巴细胞和粒细胞的数量及分布均在正常范围内。在大鼠饮食中引入所开发的乳制品后,在大鼠微生物群中观察到双歧杆菌、乳酸菌和正常大鼠微生物群典型的肠杆菌水平升高。至于表征大鼠脂质代谢的生化指标,与完整大鼠组相比,食用发酵乳制品的大鼠(对照组和处理组)血清胆固醇水平有统计学意义的降低;此外,与完整大鼠相比,食用本研究中开发的乳制品的大鼠(处理组)血清甘油三酯水平有统计学意义的降低。本研究中使用的罗伊氏乳杆菌LR1是俄罗斯纯化的第一株被鉴定可用于生产益生菌食品的菌株,因为目前市场上可见的罗伊氏乳杆菌菌株均为国外来源。

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