Strain for high yield of NAD < + > and aging resistance, microbial agent and application

Abstract

The invention discloses a bacterial strain for high yield of NAD < + > and anti-aging, a microbial agent and application, the bacterial strain is lactobacillus fermentum HLF67, is preserved in Guangdong Microbial Culture Collection Center, and has the preservation number of GDMCC NO: 65709; the preservation time is January 2, 2025; the lactobacillus mucilaginosus is separated from a breast milk source, the lactobacillus mucilaginosus can produce NAD < + > at high yield, the lactobacillus mucilaginosus is inoculated into a fermentation culture medium for fermentation treatment, the yield of NAD < + > is greatly increased, the lactobacillus mucilaginosus has excellent anti-aging performance, specifically, the content of NAD < + > in fermentation liquor reaches up to 27.6 mg / g when the lactobacillus mucilaginosus is inoculated into the fermentation culture medium, and it can be seen that the lactobacillus mucilaginosus has good application prospects. According to the technical scheme, the strain can better produce NAD < + >, the production cost of the NAD < + > is reduced, the anti-aging property is improved, and market requirements can be better met.

Description

Technical Field

[0001] The present invention relates to the technical field of NAD+ production, and in particular to a strain, a bacterial agent and applications for high-yield NAD+ and anti-aging. Background Art

[0002] NAD or NAD+ (Nicotinamide Adenine Dinucleotide) is a biological substance that transfers electrons. It is nicotinamide adenine dinucleotide in Chinese and is an oxidized coenzyme I. It is a coenzyme in many dehydrogenases in the body. Its function is to connect the tricarboxylic acid cycle and the respiratory chain, and to transfer hydrogen removed during the metabolic process to flavin proteins. Its reduced form is NADH. More precisely, NADH and H + .

[0003] The study showed that with the help of sodium ions, the Slc12a8 protein transports nicotinamide mononucleotide (NMN) directly into cells and quickly works to generate NAD. When NAD levels drop, cells also enhance the expression of the Slc12a8 gene, increasing their ability to transport xNMN.

[0004] Experimental results show that taking nicotinamide riboside can increase the content of nicotinamide adenine dinucleotide (NAD+) in the human body by 60%. NAD+ is an indispensable substance for activating acetylase and decreases with age. At present, NAD+ can be produced by strains, but the yield of NAD+ produced by existing strains is low and cannot better meet market demand.

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] The object of the present invention is to provide a strain for high-yield NAD+ and anti-aging and its application. The fermented mucus lactobacillus (Limosilactobacillus fermentum) HLF67 screened by the present invention has a high NAD+ yield, saving the NAD+ production cost.

[0007] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0008] The first aspect of the present invention provides a strain for high production of NAD+ and anti-aging, wherein the strain is Limosilactobacillus fermentum HLF67, which is deposited in Guangdong Provincial Microbiological Culture Collection Center with a deposit number of GDMCC NO: 65709; the deposit time is January 2, 2025.

[0009] The second aspect of the present invention provides a microbial agent containing the strain for high production of NAD+ and anti-aging.

[0010] Preferably, the microbial agent is obtained by inoculating the strain for high NAD+ production and anti-aging into a culture medium for cultivation.

[0011] The third aspect of the present invention provides an application of the above-mentioned strain for high production of NAD+ and anti-aging or the microbial agent in NAD+ production.

[0012] A fourth aspect of the present invention provides a method for producing NAD+ by fermentation, the method comprising the following steps:

[0013] The strain used for high NAD+ production and anti-aging is expanded and cultured, and then inoculated into a fermentation medium for fermentation culture.

[0014] Preferably, during the fermentation culture process, the pH value of the fermentation medium is controlled to be 6.0-6.5 by adding a pH regulator.

[0015] Preferably, the pH regulator is a calcium hydroxide solution with a mass volume concentration of 20%.

[0016] Preferably, the fermentation culture temperature is 35-38°C.

[0017] Compared with the prior art, the beneficial effects of the present invention include at least:

[0018] The present invention discloses a fermentation bacillus isolated from breast milk source. The fermentation mucus lactobacillus can produce a high amount of NAD+. After being inoculated into a fermentation medium and fermented, the production of NAD+ is greatly improved, and the fermentation liquid has excellent anti-aging properties. Specifically, when the fermentation medium is inoculated, the NAD+ content in the fermentation liquid is as high as 27.6 mg / g. It can be seen that the strain in the technical solution of the present application can better produce NAD+, reduce the production cost of NAD+, improve the anti-aging properties and better meet market demand.

[0019] In the present invention, NAD+ can participate in the tricarboxylic acid cycle and the respiratory chain, synthesize ATP by chemical osmotic coupling, and provide energy for cells. Adequate supply of energy helps to maintain the normal function of cells and delay the aging process. At the same time, it can also help repair damaged NDA, reduce gene mutations and cell damage, and thus delay aging. DETAILED DESCRIPTION

[0020] The following embodiments of the technical solution of the present invention are described in detail in conjunction with the embodiments. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and are therefore only used as examples, and cannot be used to limit the protection scope of the present invention.

[0021] It should be noted that, unless otherwise specified, the technical terms or scientific terms used in this application should have the common meanings understood by those skilled in the art to which the present invention belongs.

[0022] The embodiment of the present invention provides a strain for high production of NAD+ and anti-aging, wherein the strain is Limosilactobacillus fermentum HLF67, which is deposited in Guangdong Microbiological Culture Collection Center with a deposit number of GDMCC NO: 65709; the deposit time is January 2, 2025.

[0023] The above-mentioned strain of the present invention is isolated from a breast milk source. The fermented mucus lactobacillus can produce a high amount of NAD+. When inoculated into a fermentation medium and subjected to fermentation treatment, the yield of L-lactic acid quality NAD+ is greatly improved, and the production cost of NAD+ is reduced.

[0024] In the present invention, NAD+ can participate in the tricarboxylic acid cycle and the respiratory chain, synthesize ATP by chemical osmotic coupling, and provide energy for cells. Adequate supply of energy helps to maintain the normal function of cells and delay the aging process. At the same time, it can also help repair damaged NDA, reduce gene mutations and cell damage, and thus delay aging.

[0025] Another embodiment of the present invention provides a microbial agent containing the strain for high production of NAD+ and anti-aging.

[0026] In one embodiment, the microbial agent is obtained by inoculating the strain for high NAD+ production and anti-aging into a culture medium for cultivation.

[0027] Another embodiment of the present invention provides the use of the above-mentioned strain for high production of NAD+ and anti-aging or the microbial agent in NAD+ production.

[0028] Another embodiment of the present invention provides a fermentation production method of NAD+, wherein the fermentation production method of NAD+ comprises the following steps:

[0029] The strain used for high NAD+ production and anti-aging is expanded and cultured, and then inoculated into a fermentation medium for fermentation culture.

[0030] In one embodiment, during the fermentation culture process, the pH value of the fermentation medium can be controlled to any value between 6.0 and 6.5 by adding a pH regulator.

[0031] In one embodiment, the pH adjuster is a calcium hydroxide solution with a mass volume concentration of 20%.

[0032] In one embodiment, the fermentation culture temperature may be any value between 35 and 38°C.

[0033] The technical solution of the present invention is further described in detail below through specific embodiments.

[0034] Example 1

[0035] This example is the screening of Limosilactobacillus fermentum HLF67;

[0036] Five portions of breast milk from children born within 1 to 2 months were selected, and after gradient dilution, they were applied to LBS culture medium supplemented with 1% mycophenolate mofetil, and enriched and cultured in an anaerobic incubator for 60 hours. After culture, colonies were picked with an inoculation loop and streaked for purification based on the color, size, edge shape, etc. of the colonies, and 17 strains were obtained.

[0037] Example 2

[0038] This example is a study on the performance of NAD+ production by fermentation of different strains:

[0039] The 17 strains obtained in Example 1 were inoculated into LB liquid culture medium for expansion culture at a temperature of 37 degrees Celsius for 42 hours;

[0040] Then, the expanded cultured bacteria were counted and inoculated into MRS medium for fermentation at a concentration of 1×10 6 / ml, the fermentation temperature is 37°C, the pH value is controlled between 6 and 6.5 during the fermentation process, the culture time is 96h, and the fermentation liquid is obtained;

[0041] The concentration of NAD+ in the fermentation broth was detected, and the test results are shown in Table 1;

[0042] Table 1

[0043]

[0044]

[0045] From Table 1, we can see that:

[0046] The second strain had a higher production of NAD+ and was identified as fermentative mucus Lactobacillus (denoted as fermentative mucus Lactobacillus HLF67), and its morphological identification characteristics were as follows: rod-shaped, orange colonies, and irregular.

[0047] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. These modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in the scope of the claims and specification of the present invention.

Claims

1. A strain for high production of NAD+ and anti-aging, characterized in that: The strain is Limosilactobacillus fermentum HLF67, which is preserved in Guangdong Microbiological Culture Collection Center with a preservation number of GDMCC NO: 65709 and a preservation time of January 2, 2025.

2. A microbial agent containing the strain for high NAD+ production and anti-aging according to claim 1.

3. The microbial agent according to claim 2, characterized in that: The microbial agent is obtained by inoculating the strain for high NAD+ production and anti-aging into a culture medium for cultivation.

4. Use of the strain for high NAD+ production and anti-aging according to claim 1 or the microbial agent according to any one of claims 2 to 3 in NAD+ production.

5. A fermentation production method of NAD+, characterized in that: The steps include: The strain for high NAD+ production and anti-aging according to claim 1 is expanded and cultured, and then inoculated into a fermentation medium for fermentation culture.

6. The fermentation production method of NAD+ according to claim 5, characterized in that: During the fermentation culture process, the pH value of the fermentation medium is controlled to be 6.0-6.5 by adding a pH regulator.

7. The fermentation production method of NAD+ according to claim 6, characterized in that: The pH regulator is a calcium hydroxide solution with a mass volume concentration of 20%.

8. The fermentation production method of NAD+ according to claim 5, characterized in that: The temperature of the fermentation culture is 35-38°C.

Images