Lactobacillus delbrueckii subsp. bulgaricus strain with low beta-galactase activity and application thereof
A technology of the Bulgarian subspecies, Lactobacillus delbrueckii, which is applied in applications, bacteria, dairy products, etc., to achieve the effects of reducing metabolic activity, stabilizing performance, and slowing acidification
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Embodiment 1
[0038] Example 1 Screening of Lactobacillus delbrueckii subspecies KLDS1002 with low β-galactase activity
[0039] (1) screening
[0040] Fifteen strains of Lactobacillus bulgaricus isolated from traditional dairy products were activated 2-3 times and then inoculated into 11% skim milk, fermented by a single strain, and post-ripened for 24 hours to measure the β-galactosidase activity of each strain. Determination method: first take 50mL of fermented milk and mix it with 1% EDTA (pH=12, 4°C), so that the mass of each gram of sample is greater than 20mg EDTA, the main purpose is to chelate Ca through EDTA 2+ , dissolve the coagulated milk protein in the sample, then centrifuge at 5520g for 10min (4°C), remove the supernatant, wash the bacterial cells twice with 1mL of 20mmol / L phosphate buffer (pH=7.0), add 10mL of 20mmol / L phosphate buffer solution in the bacterial cells, after centrifugation at 8000r / min for 10min, the bacterial cells were collected, and then 8mL of 20mmol / ...
Embodiment 2
[0047] Example 2 Experiment on the relationship between Lactobacillus delbrueckii subsp. bulgaricus KLDS1002 with low β-galactase activity and post-acidification
[0048] (1) Determination of β-galactosidase activity
[0049] The intracellular enzymes were crudely extracted by lysozyme wall breaking method, and then the enzyme activity of β-galactosidase was determined by ONPG colorimetry. One enzyme activity unit (U) refers to the amount of enzyme required to release 1 μmol ONPG per minute at 37°C.
[0050] (2) Test the enzyme activity of β-galactosidase in the fermented milk of the test strain Lactobacillus delbrueckii subsp. bulgaricus KLDS1002 and the control commercial strain G-495 at 4°C on the 1st, 4th, 10d and 13d respectively, pH value and acidity, the results are shown in Table 2, figure 1 and figure 2 .
[0051] (3) Enzyme activity and acidity comparison measurement results
[0052] From Table 2, figure 1 and figure 2It can be seen that the post-acidificati...
Embodiment 3
[0055] Example 3 Determination of physiological characteristics of Lactobacillus delbrueckii subsp. bulgaricus strain KLDS1002
[0056] (1) Determination of growth performance
[0057] The test strain Lactobacillus delbrueckii subsp. bulgaricus KLDS1002 and the control bacteria G-495 were activated for two generations and then inoculated in MRS liquid medium at 3% and cultured at 37°C at 0, 2, 4, 6, 8, 10, Samples were taken at 12, 14, 16, 18, 20, 22, 24, 36 and 48 hours to determine the concentration of bacteria, and the results are shown in image 3 .
[0058] (2) Determination of acid resistance
[0059] Inoculate the activated Lactobacillus delbrueckii subsp. bulgaricus KLDS1002 and G-495 in MRS liquid medium. After culturing at 37°C for 16 hours, take 3% of the inoculum from each sample and inoculate them at pH 3.0 and 4.5 respectively. , 5.5, 6.5 MRS liquid medium, cultured at 37°C for 12h. The number of viable bacteria was calculated by the plate colony counting met...
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