KR-20260065076-A - Novel Lacticaseibacillus paracasei LP02 strain, and uses thereof

Abstract

The present invention relates to a novel Lacticaseibacillus paracasei LP02 strain and its uses, wherein a newly isolated and identified Lacticaseibacillus paracasei The LP02 strain completely degrades 1-kestose ( GF2 ), nystose ( GF3 ), and 1F -fructofuranosylnystose ( GF4 ), grows in a medium containing glucose or fructooligosaccharides (FOS) as a carbon source, and confirms that β -fructosidase derived from the LP02 strain degrades 1-kestose ( GF2 ), nystose ( GF3 ), and 1F - fructofuranosylnystose ( GF4 ), thereby the LP02 strain A new probiotic capable of degrading fructooligosaccharides and a health functional food composition for improving the digestion of fructooligosaccharides can be provided, and a novel β-fructosidase derived from the LP02 strain is provided as a new enzyme that completely degrades 1-kestose (GF 2 ), nystose (GF 3 ), and 1- F - fructofuranosylnystose (GF 4 ).

Inventors

• 박천석
• 김예진
• 정동현

Assignees

• 경희대학교 산학협력단
• 전남대학교산학협력단

Dates

Publication Date

20260508

Application Date

20241031

Claims (10)

1. Lacticaseibacillus paracasei, deposited under accession number KACC 92612P, which degrades fructooligosaccharides (FOS) LP02 strain.
2. A strain according to claim 1, characterized in that the fructooligosaccharide is one or more selected from sucrose (GF), 1-kestose (GF 2 ), nystose (GF 3 ), and 1F - fructofuranosylnystose (GF 4 ).
3. A strain according to claim 1, characterized in that the strain has a fosE gene consisting of a nucleotide sequence represented by SEQ ID NO. 3.
4. A strain according to claim 1, characterized in that the strain produces β-fructosidase consisting of an amino acid sequence represented by SEQ ID NO. 4.
5. Lacticaseibacillus paracasei deposited under accession number KACC 92612P β-fructosidase enzyme produced by the LP02 strain.
6. In claim 6, the enzyme is characterized by being composed of an amino acid sequence represented by SEQ ID NO. 4.
7. In claim 6, the enzyme is characterized by degrading one or more selected from sucrose (GF), 1-kestose (GF 2 ), nystose (GF 3 ), and 1F - fructofuranosylnystose (GF 4 ).
8. Lacticaseibacillus paracasei deposited under accession number KACC 92612P A functional probiotic composition containing the LP02 strain.
9. Lacticaseibacillus paracasei deposited under accession number KACC 92612P A health functional food composition for improving digestion of fructooligosaccharides containing live bacteria of the LP02 strain or a culture solution thereof as an active ingredient.
10. Lacticaseibacillus paracasei deposited under accession number KACC 92612P A feed additive containing live bacteria of the LP02 strain or a culture solution thereof as an active ingredient.

Description

Novel Lacticaseibacillus paracasei LP02 strain, and uses thereof The present invention relates to a novel Lacticaseibacillus paracasei LP02 strain and its use. The gastrointestinal tract of mammals is inhabited by a diverse array of microorganisms known as gut microbes, which play a vital role in maintaining the host's health. In particular, they are involved in nutrient absorption, immune regulation, and disease control, and interact with the host's epithelial cells and immune cells. Diet is one of the major factors influencing the composition of the gut microbial community. Consequently, dietary prebiotics are increasingly being used in human food and animal feed. As prebiotics, indigestible carbohydrates induce microbial competition, selectively stimulate the growth and activity of beneficial gut microbes, and exhibit an effect of reducing pathogens. Fructans are a group of indigestible carbohydrates with a linear structure composed of (2→1)-linked β-D-fructofuranosyl units, mostly ending in glucose residues. Fructans with short chain lengths are known as fructooligosaccharides (FOS) and are commercially used as prebiotics in foods and nutritional supplements. The global FOS market is expected to reach $7.36 billion by 2031, with inulin-type FOS being the most popular. Commercially, two different types of inulin-type FOS mixtures have been produced based on the transfructosylation process or inulin degradation. The main components of commercial FOS in inulin-type FOS are sucrose (GF), 1-kestose (GF 2 ), nystose (GF 3 ), and 1F - fructofuranosylnystose (GF 4 ). FOS that is not digested in the upper GIT of mammalian hosts reaches the large intestine and is fermented by intestinal microorganisms. During the fermentation of FOS by intestinal microorganisms, short-chain fatty acids (SCFAs), such as acetic acid, propionic acid, and n-butyric acid, are released into the intestinal lumen. SCFAs serve not only as an energy source for intestinal cells but are also major factors involved in mucosal proliferation, inflammation, colorectal cancer carcinogenesis, the removal of nitrogen compounds, and the regulation of mineral absorption. Furthermore, SCFAs are presumed to play an important role in the neuro-immunoendocrine regulation of the brain and various organs. In particular, FOS is generally considered a specific prebiotic for symbiotic lactic acid bacteria and bifidobacteria in the gut. It has been believed that FOS can selectively stimulate the growth and activity of these species. As beneficial symbiotic gut microbiota, lactic acid bacteria have been extensively studied for their use as probiotics because various species can provide health-promoting benefits, such as genitourinary health management, anti-obesity, anti-inflammatory, anticancer, and anti-allergic activities. They are also used in the production of various fermented foods. Figure 1 shows the results of thin-layer chromatography (TLC) analysis of the degradation patterns of 13 types of lactic acid bacteria isolated from fecal samples against fructooligosaccharides (FOS). G represents glucose, F represents fructose, S represents sucrose, and MRS represents the medium without the strains inoculated. Figure 2 shows Lacticaseibacillus paracasei in a medium containing glucose or fructooligosaccharides (FOS) as a carbon source This is the result of analyzing the growth rate and carbon source degradation pattern of the LP02 strain by TLC. G represents glucose, F represents fructose, and S represents sucrose; the black circles represent the results in a medium containing FOS as a carbon source, and the white circles represent the results in a medium containing glucose as a carbon source. Figure 3 shows Lacticaseibacillus paracasei in a medium containing fructooligosaccharides (FOS) as a carbon source LP02 strain and Lacticaseibacillus paracasei This is the result of analyzing the degradation pattern of FOS by TLC after inoculating the KCTC 3510 T strain. G represents glucose, F represents fructose, and S represents sucrose; Lane 1 is the result for the LP02 strain, and Lane 2 is the result for the KCTC 3510 T strain. Figure 4 shows Lacticaseibacillus paracasei LP02 strain and Lacticaseibacillus paracasei The FOS operon of the KCTC 3510 T strain is the result of comparative analysis. The numbers above/below the gene and in parentheses indicate the chromosomal location and gene size, respectively. The percentage between genes indicates nucleotide similarity. Figure 5 shows Lacticaseibacillus paracasei This is the result of analyzing the top 20 genes that are upregulated in the presence of FOS in the LP02 strain. Expression levels were expressed as log 2 fold changes (log 2 FC), and the alphabet (ae) grouped before the Locus tag indicates that the genes were clustered together. Figure 6 shows Lacticaseibacillus paracasei This is the result of analyzing the FOS degradation ability by TLC after constructing a recombinant protein (rFosE) of β-fructosidase der