Lactobacillus plantarum LLY-606 and application thereof
A technology of LLY-606 and Lactobacillus plantarum, applied in the direction of application, bacteria, microorganisms, etc., can solve the problems of low acceptance, achieve strong acid resistance and bile salt resistance, reduce cholesterol and triglycerides, and excellent probiotic effects Effect
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Embodiment 1
[0044] In this example, Lactobacillus plantarum LLY-606 was isolated from Inner Mongolia farmhouse fermented yoghurt food with an optimized medium, and its identification and bacterial safety test were carried out. The details are as follows:
[0045] (1) Medium formula
[0046] In this example, optimized MRS solid medium and MRS culture medium were prepared.
[0047] MRS solid medium, referred to as MRS medium, the formula for 1L is: casein peptone 10.0g / L, beef extract 10.0g / L, yeast extract 5.0g / L, glucose 20.0g / L, dipotassium hydrogen phosphate 2.0g / L, Tween 80 1.0g / L, triammonium citrate 2.0g / L, sodium acetate 5.0g / L, magnesium sulfate 0.1g / L, manganese sulfate 0.05g / L, agar 17.5g
[0048] MRS broth medium, that is, the formula of MRS culture medium is only that agar is not added to MRS medium, and the rest are the same.
[0049] The prepared medium and culture solution were adjusted to pH 6.5, sterilized at 121°C for 20 minutes, and then refrigerated at 4°C for use. ...
example 16
[0052] The primer sequences for 16s DNA amplification in this example are shown in SEQ ID NO.1 and SEQ ID NO.2.
[0053] SEQ ID NO.1: 5'-AGAGTTTGATCATGGCTCAG-3'
[0054] SEQ ID NO.2: 5'-TAGGGTTACCTTGTTACGACTT-3'
[0055] Primers were synthesized by Shenzhen BGI Institute.
[0056] The PCR reaction system is, 50 μL reaction solution includes: 1 μL of 10 mM dNTPs, 5 μL of 10× buffer, 10 μL of 10 mM upstream and downstream primers, 5 μL of bacterial liquid PCR template, 1 μL of 5U / μL Taq enzyme, supplemented with ddH 2 O supplemented to 50 μL.
[0057] The PCR reaction conditions are: 95°C pre-denaturation for 5 minutes, and then enter 35 cycles: 95°C for 45s, 55°C for 45s, 72°C for 30s, after the cycle ends, extend at 72°C for 5min, and stand by at 4°C.
[0058] The PCR amplification products were recovered by gel cutting, and the recovered PCR products were sequenced. In this example, the kit TaKaRa MiniBEST agarose GeL DNA Extraction kit was used for gel extraction and rec...
Embodiment 2
[0087] In this example, Lactobacillus plantarum LLY-606 isolated in Example 1 was tested for cholesterol-lowering function in vitro, and the standard strain Lactobacillus plantarum 299v purchased from CGMCC was used as a control. Among them, Lactobacillus plantarum 299v is a commercially available strain with cholesterol-lowering function. The specific test is as follows:
[0088] First prepare the MRS culture solution containing cholesterol (abbreviated as MRSO-CHOL culture solution) and the MRS culture solution without cholesterol.
[0089] MRSO-CHOL culture medium: composed of MRS culture medium, sodium thioglycolate, bile salt and cholesterol, wherein the concentration of sodium thioglycolate in MRSO-CHOL culture medium is 2g / L, and the concentration of bile salt in MRSO-CHOL culture medium The concentration is 0.3% by weight, and the concentration of cholesterol is 120 μg / mL.
[0090] Lactobacillus plantarum LLY-606 and Lactobacillus plantarum 299v were respectively tra...
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