Novel lactic acid bacteria and its applications in immunomodulation and anti-inflammation
A kind of lactic acid bacteria, anti-inflammatory technology, applied in the field of lactic acid bacteria and compositions comprising the same
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example 1
[0039] Example 1 Isolation and gene classification of Lactobacillus plantarum subsp.Plantarum K21
[0040] Lactobacillus subspecies germ K21 is isolated from Taiwan's traditional fermented food Fucai. The 16S rRNA gene and pheS gene DNA of K21 were analyzed by direct sequencing of PCR amplified DNA fragments.
[0041]PCR was performed according to the conditions described in the table below, using 16S rRNA gene primers (Bact-8F (II): 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID NO: 3)); 15R: 5'-AAGGAG GTGATCCAACCGCA-3' (SEQ ID NO :4) or phe primer (pheS-forward: 5'-CAYCCNGCHCGYGAYATGC-3' (SEQ ID NO:5; pheS-reverse: 5'-CCWARVCCRAARGCAAARCC-3' (SEQ ID NO:6)) for PCR. The obtained Sequences were input into the sequence alignment software provided by the National Center for Biotechnology Information (NCBI) (http: / / www.ncbi.nlm.nih.gov / ) on the Internet for manual sequence alignment, and Align with representative pheS gene DNA sequences of organisms belonging to Firmicutes. For comparison,...
example 2
[0055] Example 2 Identification of bacterial strains using RAPD-PCR
[0056] K21 was compared with RAPD markers of two other Lactobacillus plantarum strains (ATCC14917T and ATCC17638T). Under the conditions listed in Table 3, PCR was performed using random primer 1254 (5'-CCGCAGCCAA-3', SEQ ID NO: 7). DNA extracted from each of these strains was used as a template. The resulting amplified products were subjected to electrophoresis and compared to patterns, such as figure 1 shown. This result shows that K21 has a specific PCR fingerprint derived from its genome, representing that K21 is a new strain.
[0057] Table 3: Composition of PCR reaction solution (25 μl)
[0058] ingredients
[0059] PCR conditions: 94°C, 2 minutes; 5 cycles (94°C, 30 seconds; 36°C, 1 minute; 72°C, 1.5 minutes); 30 cycles (94°C, 20 seconds; 36°C, 30 seconds; 72°C, 1.5 minutes); 72°C, 3 minutes.
example 3
[0060] Example 3 Analytical Profile Index (Analytical Profile Index; API) Typing
[0061] The sugar utilization rate of K21 used in the present invention was studied using the API50CHL kit (bioMerieux, France), and the results are shown in Table 4. Fermentation tests indicated that K21 had biochemical properties similar to Lactobacillus plantarum subsp. germ.
[0062] Table 4: Results of fermentation tests a
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PUM
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