Lactobacillus plantarum for reducing antigenicity of beta-lactoglobulin of cow milk and application thereof
A technology of Lactobacillus plantarum and lactoglobulin, applied in the field of Lactobacillus plantarum AHQ-14
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Embodiment 1
[0021] Embodiment 1: Lactobacillus plantarum AHQ-14 screening
[0022] I. strain isolation method: prepare MRS medium: according to 20g / L glucose, 10g / L peptone, 5g / L yeast extract powder, 10g / L beef extract, 5g / L sodium acetate, 2g / L diamine hydrogen citrate, 2g / L dipotassium hydrogen phosphate, 0.58g / L MgSO 4 ·7H 2 O, 0.25g / L MnSO 4 ·5H 2 O and 1mL / L Tween-80 Weigh the raw materials, mix them evenly, dissolve them in deionized water, and sterilize the obtained solution for 15 minutes at a temperature of 121°C to obtain the MRS medium; add 15g / L agar powder to the medium , and sterilized at a temperature of 121° C. for 15 minutes to obtain an MRS agar medium.
[0023] The collected Xinjiang traditional fermented yoghurt sample was mixed with a vortex oscillator, 0.5g or 0.5ml sample was weighed and put into 4.5ml concentration of 0.85% sterilized physiological saline, and the sample was diluted tenfold (10 -4 , 10 -5 , 10 -6 ), then smeared on MRS agar medium or M17 ag...
Embodiment 2
[0025] Example 2: Morphological characteristics and strain identification of Lactobacillus plantarum AHQ-14
[0026] I. Colony Morphological Characteristics: Lactobacillus plantarum AHQ-14 of the present invention forms milky white colonies on MRS agar medium, opaque, round, neat edges, central bulge, about 1.3~1.6mm in diameter, specifically see appendix figure 1 .
[0027] II. Morphology of bacteria: Lactobacillus plantarum AHQ-14 of the present invention has the following morphological characteristics: the cells are long rod-shaped, with blunt ends, 4.0-8.0 μm in length, and 0.9-1.1 μm in width, and are single or chain-shaped. For details, see attached figure 2 .
[0028] III. Molecular biological identification: Universal primers are used, the forward primer is 27f: 5′-AGAGTTTGATCCTGGCTCAG-3′; the reverse primer is 1492r: 5′-GGTTACCTTGTTACGACTT -3′, and the bacterial DNA is used as the amplification template for PCR amplification The 16S rRNA gene region, the amplified...
Embodiment 3
[0030] Embodiment 3: the bile salt hydrolyzing enzyme activity of plantarum lactobacillus AHQ-14
[0031] The strain was supplemented with 0.2% (w / v) sodium thioglycolate, 0.3% (w / v) sodium taurodeoxycholate hydrate and 0.2% (w / v) CaCl 2 cultured on MRS agar plates. Add the above-mentioned sterilized MRS medium into a sterile petri dish, and evenly place sterile filter paper sheets with a diameter of about 3-4 mm on the solidified medium. Adsorb 10 μL of activated bacterial liquid, slowly add to filter paper until the liquid no longer flows, and culture at 37°C for 3 days under anaerobic conditions. The precipitated area around the colony shows the bile salt hydrolase activity of the strain.
[0032] MRS culture medium was used as a negative control, and LGG was used as a positive control for the qualitative determination of bile salt hydrolytic enzyme activity. Figure 4 A large amount of deoxycholic acid precipitated and diffused into the medium around AHQ-14, which prove...
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