CN111184133A - Pet health product and preparation method thereof - Google Patents

Abstract

The application provides a pet health product which comprises β -1, 0.01-50% of 3-glucan and 0.01-50% of nicotinamide mononucleotide in percentage by weight, and the pet health product is beneficial to improving intestinal tracts, improving cell activity, promoting metabolism and delaying senescence.

Description

Pet health product and preparation method thereof

Technical Field

The application relates to the technical field of pet foods, in particular to a pet health product and a preparation method thereof.

Background

With the increase of economy, the pet industry has been rapidly developed, and the pet food on the market is also good and uneven. Because of the quality of the pet food, the nutrition intake of the pet is determined and is related to the health of the pet; good diet is essential to ensure the health of pets, and the choice of food items for good pets is very critical to them. Research shows that pets with aging and low immunity are easy to be infected with diseases, and in order to improve the immunity of pets, special pet food is required to be fed to the pets except for paying attention to sanitary feeding habits. At present, the traditional pet formula food or formula food lacks products with real efficacy, and most of the traditional antioxidant raw materials such as vitamin E and the like are taken as main materials; or the raw materials surrounding the addition of various trace elements. Therefore, the development of the pet health care product which is beneficial to delaying senility and enhancing immunity is of great significance.

Disclosure of Invention

In order to solve the above technical problems, the present application provides a pet health product having effects of improving intestinal tracts, increasing cell activity, promoting metabolism, and delaying aging, and a method for preparing the same.

In a first aspect, the present application provides a pet health product comprising β -1, 3-glucan 0.01-50% and Nicotinamide Mononucleotide (NMN) 0.01-50% by weight.

Optionally, the mass ratio of β -1,3 glucan to nicotinamide mononucleotide is 1 (1-100).

Optionally, the mass ratio of β -1,3 glucan to nicotinamide mononucleotide is 1 (1-50), further optionally, the mass ratio of β -1,3 glucan to nicotinamide mononucleotide is 1 (2-10), for example, the mass ratio of β -1,3 glucan to nicotinamide mononucleotide is specifically 1 (1-5), 1 (1-10), 1 (2-30), 1 (1-40), 1 (1-50), 1 (1-60), 1 (1-70), 1 (1-80), 1 (1-90) or 1 (1-100).

According to the pet health product composition, intestinal health can be effectively maintained, β -1,3 glucan and nicotinamide mononucleotide in the pet health product play a synergistic effect, β -1,3 glucan can obviously improve an antioxidant index, so that immunity of a pet is improved, aging is delayed, nicotinamide mononucleotide provides an energy source ATP for cells, metabolism and cell renewal are enhanced, absorption and function implementation of glucan are facilitated, the nicotinamide mononucleotide also has an aging delaying function, the function is improved after being matched with glucan, and the whole pet health product can show a more prominent anti-aging effect.

Specifically, β -1,3 glucan can induce the proliferation, secretion and phagocytic activity of macrophages, wherein the induced IL-1 (interleukin-1) can indirectly stimulate Th cells (helper T cells) to secrete anticancer cytokines such as IL-2 (interleukin-2), and the like, the induction of the cytokines can start a series of biological reactions, such as IL-1 promoting the proliferation and activation of cells such as T, B, NK and the like, IL-2 inducing LAK (killer cell activated by lymphokine) and TIL (tumor infiltrating lymphocyte) and the like to kill pathogenic microorganisms, β -1, 3-glucan has an obvious bidirectional characteristic on the regulation of immune response, namely shows positive regulation (immune enhancement) under the condition of adaptive dosage, but shows negative regulation (immune inhibition) under the condition of excessive dosage.

In the present application, nicotinamide mononucleotide is NAD⁺(nicotinamide adenine dinucleotide) synthesized precursor, after entering cells, synthesizes NAD through a one-step metabolic process⁺It can improve the vitality and immunity of cells by promoting the cells to produce Adenosine Triphosphate (ATP).

Optionally, the nicotinamide mononucleotide is more than 99% pure the β -1,3 glucan is more than 70% pure, for example, the β -1,3 glucan is 70% -95% pure.

Optionally, further active components are included, the further active components including at least one of vitamins, antioxidants and minerals.

Optionally, the vitamins include one or more of vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, and vitamin E; the antioxidant comprises one or more of Butylated Hydroxyanisole (BHA), dibutyl hydroxy toluene (BHT), Propyl Gallate (PG), D-sodium erythorbate and tea polyphenol; the mineral comprises mineral elements including one or more of magnesium, iron, zinc, copper, calcium, iodine and manganese.

Optionally, in the mineral, magnesium can be derived from one or more of magnesium sulfate, magnesium chloride and magnesium citrate; the iron can be one or more of ferrous gluconate, ferrous sulfate, ferrous lactate, ferric citrate and ferric pyrophosphate; the zinc can be one or more of zinc sulfate, zinc gluconate, zinc lactate and zinc chloride; the copper may be derived from copper sulfate; the calcium can be selected from one or more of milk calcium, calcium citrate, calcium phosphate, calcium chloride, calcium carbonate and calcium lactate; iodine may be derived from potassium iodide; the manganese may be derived from manganese sulfate.

Optionally, the pet care product further comprises one or more of a filler, an excipient, and a preservative. Optionally, the bulking agent comprises one or both of a plant starch and a dextrin; the excipient comprises one or more of calcium carbonate, sucrose powder, lactose powder and pectin; the preservative comprises one or more of potassium sorbate, sodium benzoate, calcium propionate, and sodium propionate.

Optionally, in the daily dosage of the pet health product, the β -1, 3-glucan is 1-1000 micrograms/kg of body weight, and the nicotinamide mononucleotide is 1-2000 micrograms/kg of body weight.

The pet health product can be in the form of capsules and can also be in the form of pet feed.

In a second aspect, the present application also provides a method for preparing a pet health product according to the first aspect of the present application, comprising:

weighing the components according to the formula proportion, putting the components into a stirring pot, and stirring to obtain a mixed material; and then, after forming and drying, cooling to room temperature, and then accommodating in a packaging container and sealing to obtain the pet health product.

Optionally, after the mixture is obtained after the stirring, before the molding, the grinding of the mixture is further performed. Wherein, the particle size of the mixed material after grinding treatment can be adjusted based on practical application. In one embodiment, the particle size of the mixture after the grinding process is 100-.

Alternatively, the packaging container may be a capsule, and the packaging container may also be other forms of packaging. For example, the pet care product described herein is shown in fig. 1.

Optionally, the β -1,3 glucan is prepared by a fermentation process, and the specific fermentation process comprises:

preparing culture solution, inoculating zymocyte into the culture solution, fermenting at 28-32 deg.C until OD600 value of the culture solution is greater than 0.6-0.9, separating and extracting β -1,3 dextran, wherein the culture solution contains potassium dihydrogen phosphate (KH)₂PO₄) Sodium nitrate (NaNO)₃) Magnesium sulfate (MgSO)₄) Calcium chloride (CaCl)₂) Ferrous chloride (FeCl)₂) Sucrose and agar.

Optionally, the mass fractions of the components in the culture solution are: KH (Perkin Elmer)₂PO₄1‰，NaNO₃3‰，MgSO₄0.2‰，CaCl₂0.07‰，FeCl₂0.0125 per mill, 20 per mill of cane sugar and 0.9 percent of agar. The "% o" here is a thousandth, representing a few thousandths of a word.

Optionally, the pH of the culture broth is 7.0-7.2. The activity of the zymophyte in the pH range is more prominent, and the fermentation efficiency and the fermentation quality are improved.

Alternatively, the fermenting bacteria may be, but not limited to, Agrobacterium ZX09(Agrobacterium sp. zx09).

Continuously, in the fermentation process, the culture solution inoculated with the zymophyte is continuously stirred, and air is introduced, wherein the stirring speed is 100-300rpm, and the ventilation volume is 0.3-0.5 vvm.

Optionally, the fermentation time can be adjusted based on the OD600 value of the culture solution during the fermentation process. In one embodiment, the fermentation time is 16-30 hours.

Further, optionally, during the fermentation process, the fermentation temperature may be specifically 30 ℃.

According to the application, β -1,3 glucan with the purity of more than 70% can be obtained after β -1,3 glucan prepared by fermentation is separated and extracted, since β - (1,3) glucan is usually extracted from shiitake mushroom, lucid ganoderma, yeast or microalgae in the prior art, a large amount of raw materials are consumed, the yield is very low, the purity is low, and the production cost is very high, secondly, a large amount of acid, alkali and organic solvent are applied in the extraction process, so that a large amount of waste water is unfavorable to the environment, meanwhile, β - (1,3) glucan from yeast and microalgae is not water-soluble, the biological activity is low, compared with the prior β -1,3 glucan preparation process, β - (1,3) glucan prepared by the method has the characteristics of high yield and high purity, and the raw material cost of β - (1,3) glucan can be greatly reduced.

Optionally, the method for synthesizing nicotinamide mononucleotide comprises:

constructing a biological enzyme recombinant plasmid, and performing protein expression and purification to obtain biological enzymes, wherein the biological enzymes comprise a first biological enzyme, a second biological enzyme, a third biological enzyme and a fourth biological enzyme; the first biological enzyme comprises the amino acid sequence as set forth in SEQ ID NO:1 and the second biological enzyme comprises the nucleotide sequence shown as SEQ ID NO: 2 and the third biological enzyme comprises the nucleotide sequence shown as SEQ ID NO: 3 and the fourth biological enzyme comprises the nucleotide sequence shown as SEQ ID NO: 4;

preparing a reaction solution, wherein the reaction solution contains inosine, adenosine diphosphate and adenosine triphosphate, adding the first biological enzyme, the second biological enzyme and the third biological enzyme into the reaction solution, generating 5-phosphoribosyl pyrophosphate after biocatalysis, adding nicotinamide and the fourth biological enzyme, and separating to obtain nicotinamide mononucleotide after biocatalysis.

Alternatively, the vector plasmid used for the recombinant plasmid of the biological enzyme is pET22b (+). The specific process of preparing the biological enzyme can be as follows: after the biological enzyme recombinant plasmid is transfected into the microbial strain, inducing the microbial strain containing the biological enzyme recombinant plasmid in a microbial culture solution to express the biological enzyme. The microbial strains comprise one or two of Rosetta (DE3) Escherichia coli and BL21(DE3) Escherichia coli. Further optionally, the microbial strain comprises Rosetta (DE3) escherichia coli.

The biological enzyme may be produced intracellularly by the microorganism strain, or may be produced intracellularly by the microorganism strain and then released into the culture medium of the microorganism. And collecting the microbial strains capable of expressing the biological enzyme, cleaning, breaking cells and collecting to obtain the crude enzyme solution, wherein the crude enzyme solution contains the biological enzyme. Then obtaining the biological enzyme after the subsequent protein purification process. In one embodiment, the purified biological enzyme protein may be obtained by nickel column purification.

Optionally, the first biological enzyme comprises a sequence as set forth in SEQ ID NO: 5. The second biological enzyme comprises the amino acid sequence shown as SEQ ID NO: 6. The third biological enzyme comprises the amino acid sequence as shown in SEQ ID NO: 7. The first biological enzyme comprises the amino acid sequence as set forth in SEQ ID NO: 8.

Alternatively, the genes encoding the biological enzymes including the first biological enzyme, the second biological enzyme, the third biological enzyme and the fourth biological enzyme should take into account degenerate bases. Taking the first biological enzyme as an example, the coding gene of the amino acid sequence shown in SEQ ID NO. 5 comprises the nucleotide sequence shown in SEQ ID NO. 1, and the protection scope should also protect the nucleotide sequence with base degeneracy with the SEQ ID NO. 1, and the corresponding amino acid sequence of the nucleotide sequence is still SEQ ID NO. 5.

The gene encoding the biological enzyme may be inserted into a multiple cloning site of a vector plasmid, for example, between the sites of BamHI, EcoRI or Xho I. When the coding gene of the biological enzyme is inserted into a pET22b (+) vector plasmid, an initiation codon (such as ATG) can be added at the 5' end of the coding gene of the biological enzyme; a stop codon (e.g., TAA) may be added to the 3' end. In one embodiment, the gene encoding the biological enzyme may be inserted between the cleavage sites of "CATATG" and "CTCGAG" of the vector plasmid.

Optionally, the reaction liquid further comprises sodium carbonate, magnesium chloride and potassium chloride. Sodium carbonate, magnesium chloride and potassium chloride in the reaction solution can be used for assisting enzyme catalysis reaction, and the activity of the biological enzyme is improved.

The nicotinamide mononucleotide synthesized by using the biological enzyme catalysis is high in yield and purity; the cost can be greatly saved; the purity of the prepared nicotinamide mononucleotide can reach more than 99%.

The beneficial effects of the application comprise the following aspects:

1. the pet health product can effectively maintain intestinal health, β -1,3 glucan and nicotinamide mononucleotide in the pet health product play a synergistic effect, β -1,3 glucan can obviously improve the level of short-chain fatty acid in cecum and improve the health level of intestinal tract, so that immunity of pets delays aging, nicotinamide mononucleotide provides energy source ATP for cells, metabolism and cell renewal are enhanced, absorption and function implementation of glucan are facilitated, nicotinamide mononucleotide has the function of delaying aging, the function is improved after being matched with glucan, and the whole pet health product can show more prominent anti-aging effect.

2. The preparation method of the pet health product is simple in process, environment-friendly and widely suitable for industrial mass production, and the β -1, 3-glucan and nicotinamide mononucleotide in the raw materials is low in preparation cost and high in purity, so that the production cost of the pet health product can be further reduced, and the anti-aging effect of the pet health product is improved.

Advantages of the present application will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the embodiments of the present application.

Drawings

In order to more clearly explain the content of the present application, the following detailed description is given in conjunction with the accompanying drawings and specific embodiments.

FIG. 1 is a schematic view of a pet health product provided in an embodiment of the present application;

FIG. 2 is a SDS-PAGE pattern of a first biological enzyme according to an embodiment of the present application;

FIG. 3 is an SDS-PAGE pattern of a second biological enzyme according to an embodiment of the present application;

FIG. 4 is an SDS-PAGE pattern of a third biological enzyme according to an embodiment of the present application;

FIG. 5 is an SDS-PAGE pattern of a fourth biological enzyme according to an embodiment of the present application;

FIG. 6 is a diagram of a specific bio-enzyme catalytic process pathway of nicotinamide mononucleotide, provided in an embodiment of the present application.

Detailed Description

While the following is a preferred embodiment of the embodiments of the present application, it should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the embodiments of the present application, and such improvements and modifications are also considered to be within the scope of the embodiments of the present application.

Unless otherwise specified, the raw materials and other chemical agents used in the examples of the present application are commercially available.

(1) β preparation of 1, 3-glucan

The β -1, 3-glucan is prepared by fermentation method, and the method comprises preparing culture solution, inoculating zymocyte ZX09 into the culture solution, wherein the culture solution is KH₂PO₄1‰，NaNO₃3‰，MgSO₄0.2‰，CaCl₂0.07‰，FeCl₂0.0125 per mill, 20 per mill of cane sugar and 0.9 percent of agar, wherein the pH value of the culture solution is 7.0-7.2, the culture solution is fermented in a fermentation tank at the temperature of 30 ℃, the stirring speed is 200rpm, the ventilation quantity is 0.4vvm, the fermentation is carried out for 1 day until the OD600 value of the culture solution is more than 0.9, and β -1,3 glucan obtained by separation and extraction is obtained, wherein the purity of the extracted β -1,3 glucan is more than 70%.

(2) Preparation of nicotinamide mononucleotide

(a) Construction of pET22b (+) -NMN recombinant plasmid

The gene sequence of the biological enzyme is synthesized by the whole gene, the biological enzyme comprises a first biological enzyme, a second biological enzyme, a third biological enzyme and a fourth biological enzyme, and the gene sequences are respectively shown as SEQ ID NO: 1-SEQ ID NO: 4, respectively and correspondingly constructing a plasmid pET22b (+), wherein a biological enzyme gene of the nucleotide sequence is inserted between Nde I and Xho I enzyme cutting sites in the vector plasmid; when the biological enzyme gene is inserted into a vector plasmid, a stop codon (such as TAA) can be added at the 3' end of the coding sequence of the biological enzyme gene to be connected with an XhoI enzyme cutting site in the vector plasmid.

(b) Expression of biological enzymes

Escherichia coli containing the recombinant plasmid pET22b (+) -NMN was inoculated at 1% inoculum size into a 50mL Erlenmeyer flask containing 10mL LB medium (100. mu.g/mL ampicillin (Amp)), maintained at 37 ℃ with a constant shaking rate of 200rpm, overnight culture was performed, then the inoculum size of 1% inoculum size was transferred to a 2L Erlenmeyer flask containing 1L LB medium (100. mu.g/mL Amp), incubation was continued at 37 ℃ until the OD600 value in the medium reached about 0.6, isopropyl- β -D-thiogalactoside (IPTG) inducer (final concentration of 0.5mM) was added, the cells were cultured at 37 ℃ for 8 hours, centrifuged and collected, then resuspended in 4-fold mass of 100mM potassium phosphate buffer (pH 6.5) and sonicated for 20min to obtain a crude NMN-synthesizing bio-enzyme solution.

And (3) carrying out protein purification on the NMN biological enzyme crude enzyme solution obtained by expression. Purifying the obtained various biological enzymes by a nickel column separation and purification method, and eluting by using imidazole with different concentrations; and then identified by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), see FIGS. 2-5. Wherein the molecular weights of the first biological enzyme, the second biological enzyme, the third biological enzyme and the fourth biological enzyme are respectively about 32KDa, 21KDa and 58 KDa. The biological enzyme protein obtained by nickel column purification is preserved for standby.

(c) Catalytic synthesis of nicotinamide mononucleotide

Preparing reaction liquid, taking inosine, adenosine diphosphate, adenosine triphosphate and nicotinamide as raw materials, taking sodium carbonate, magnesium chloride and potassium chloride as auxiliary materials, and preparing the nicotinamide mononucleotide under the catalysis of biological enzymes, wherein the synthesis route is shown in figure 6, inosine in the reaction liquid generates 1-phosphoribosyl under the catalysis of first biological enzymes, and then 1-phosphoribosyl, adenosine diphosphate and adenosine triphosphate generate 5-phosphoribosyl pyrophosphate under the catalysis of second biological enzymes and third biological enzymes. The obtained 5-phosphoribosyl pyrophosphate and the added nicotinamide generate nicotinamide mononucleotide under the catalysis of fourth biological enzyme, and then the nicotinamide mononucleotide is obtained by separation and extraction. The purity of the nicotinamide mononucleotide can reach more than 99%.

Example 1

A preparation method of a pet health product comprises the following steps:

β -1, 3-glucan 5mg, nicotinamide mononucleotide 10mg and vitamin C3 mg prepared by the preparation method are uniformly mixed and then filled into capsules to obtain the pet health care product.

Example 2

A preparation method of a pet health product comprises the following steps:

β -1, 3-glucan 5mg, nicotinamide mononucleotide 10mg and antioxidant D-sodium erythorbate 0.5mg prepared by the preparation method are uniformly mixed and then filled into capsules to obtain the pet health care product.

Example 3

A preparation method of a pet health product comprises the following steps:

β -1, 3-glucan 5mg, nicotinamide mononucleotide 10mg and mineral 0.2mg prepared by the preparation method are uniformly mixed and then filled into capsules to obtain the pet health care product.

Effects of the embodiment

(1) Regulation and control of dog serum antioxidant enzyme activity and alleviation of oxidative damage by pet health product

The material and the method are characterized in that 40 adult dogs (older than 5 years) are selected and randomly divided into 5 treatment groups, 10 treatment groups are selected in each group, the average age and the weight of each group of dogs are close to 30-35 kg.5 treatment groups, (1) a control group 1 is normal dog food, (2) a control group 2 is normal dog food and β -1,3 glucan, (3) a control group 3 is normal dog food and nicotinamide mononucleotide, the health care product is administrated by gastric lavage for 30 days and 60 days, specifically referring to table 1, forelimb vein blood collection and serum centrifugation are carried out, 4) an experimental group 1 is normal dog food and pet health care products, (5) an experimental group 2 is normal dog food and high-content pet health care products, and the serum oxidation resistance index measurement comprises glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) activity and Malondialdehyde (MDA) content.

Wherein, data statistics and analysis: single-factor analysis of variance was performed using SPSS 2.0 statistical software and multiple comparisons were performed using Duncan, with P <0.05 indicating significant differences, and the results are shown in table 2.

TABLE 1 feed rates (mg/kg BW) for various treatment groups for various components of pet health care product

TABLE 2 Effect of Pet health products on canine serum antioxidant enzyme activity and MDA content

Note: the differences (a, b, c) among the same row of shoulder letters indicate significant differences (P < 0.05).

As can be seen from the data in Table 2, the activity of GSH-Px and SOD enzyme in the canine serum of experimental group 1-2 is significantly higher than that of the control group 1-3(P <0.05), and the activity of GSH-Px and SOD enzyme in the canine serum of control group 1 and control group 3 is close, indicating that the activity of nicotinamide mononucleotide on GSH-Px and SOD enzyme is not significant, the activity increasing effect of β -1,3 glucan in control group 2 on GSH-Px and SOD enzyme is limited, significantly lower than that of experimental group 1 and 2(P <0.05), and the trend of results of 30 days and 60 days is the same, meanwhile, the MDA content in the canine serum of experimental group 1-3 is significantly lower than that of experimental group 1 and 2(P <0.05), and the canine serum content of experimental group 1 and 2 is also significantly lower than that of control group 2 and 3(P <0.05), the trend of results of 30 days and 60 days is the same, the canine serum MDA content in experimental group 1 and 2 of this embodiment is significantly lower than that of the antioxidant lipid-increasing effect of nicotinamide mononucleotide in canine serum β, and the antioxidant effect of nicotinamide mononucleotide can be further increased by the synergistic effect of antioxidant lipid-antioxidant effect of the antioxidant lipid-increasing the antioxidant lipid-antioxidant effect of the antioxidant lipid-antioxidant effect of the antioxidant lipid-lipid.

As can be seen from the data in Table 2, compared with the control group 1, the activities of GSH-Px and SOD enzyme in the serum of the dog in the control group 2 are increased to a certain extent, the MDA content is reduced (P <0.05), the activities of GSH-Px, SOD enzyme and MDA content in the serum of the dog in the nucleotide group are not obviously changed, the results of 30 days and 60 days are consistent, which shows that β -1 and 3 glucan fed alone have certain antioxidation, and nicotinamide mononucleotide fed alone has no obvious antioxidation, but the activities of GSH-Px and SOD enzyme in the serum of the dog in the experiment group 1 are obviously higher than those of the control group 2, and the MDA content is obviously lower than that of the control group 2(P <0.05), which shows that β -1,3 glucan and nicotinamide mononucleotide in the pet health care product in the experiment group 1 of the application have synergistic effect, and can greatly improve the antioxidation capability of the dog.

In the experimental group 1 and the experimental group 2 of the embodiment, the pet health product can be fed for more than 30 days to remarkably improve the activity of the antioxidant in the serum of the dog and reduce the content of lipid peroxide in the blood.

The pet health product composition can effectively maintain intestinal health, β -1,3 glucan and nicotinamide mononucleotide in the pet health product play a synergistic effect, β -1,3 glucan can obviously improve an antioxidant index, so that immunity of a pet is improved, and aging is delayed, the nicotinamide mononucleotide provides an energy source ATP for cells, metabolism and cell renewal are enhanced, absorption and function implementation of glucan are further facilitated, the nicotinamide mononucleotide has an aging delaying function, the function is improved after being matched with glucan, and the whole pet health product can show a more prominent anti-aging effect.

The above-mentioned embodiments only express several embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present application. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, which falls within the scope of protection of the present application. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Sequence listing

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Lichenwu tea

Jili Hua

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ctgcaagtta gcggtccact gctggtggtt cagctcctcg aaacgccact gctgtgtctg 420

gttagctatg cgagtctggt tgcgacgaat gcggcccgtc tgcgtctgat tgcgggtccg 480

gaaaagcgtc tgctggagat gggtctgcgt cgcgcccaag gtccagatgg cggtctcacc 540

gccagtacgt acagctatct gggcggcttt gatagcagta gcaatgttct ggcgggccag 600

ctccgtggtg ttccagttgc gggtacgctg gcccatagct tcgttaccag tttcagcggc 660

agcgaagtgc cgccagaccc aatgctggcc ccggccgcgg gtgaaggccc gggcgttgat 720

ctggcggcca aagcccaagt ttggctggaa caagtttgcg cgcatctcgg tctgggcgtt 780

caagaaccgc atccgggcga acgtgccgcc tttgtggcgt acgcgctggc ctttccacgt 840

gcctttcaag gtctgctgga tacctacagc gtttggcgta gtggtctccc aaatttcctc 900

gcggttgcgc tggcgctggg tgaactcggt taccgcgccg ttggtgttcg cctcgatagc 960

ggtgatctgc tccagcaagc ccaagaaatt cgcaaagtgt ttcgcgccgc ggccgcccag 1020

tttcaagtgc catggctgga aagcgttctg atcgtggtga gtaacaatat cgacgaagaa 1080

gcgctcgcgc gtctggcgca agaaggtagc gaggtgaacg tgatcggcat tggcaccagc 1140

gttgttacgt gcccacagca gccaagtctc ggtggcgtgt ataaactggt tgcggttggt 1200

ggccagccgc gcatgaaact gaccgaggac ccggaaaaac aaacgctgcc gggcagcaaa 1260

gccgcgtttc gtctgctggg cagcgatggt agtccgctca tggatatgct ccagctggcg 1320

gaagaaccag ttccgcaagc gggtcaagaa ctgcgtgttt ggccaccggg cgcgcaagaa 1380

ccatgcacgg ttcgtccagc ccaagttgag ccgctgctcc gtctgtgtct gcaacaaggc 1440

caactgtgcg agccgctgcc gagtctggcg gaaagtcgtg cgctcgcgca gctgagtctg 1500

agccgtctca gcccggaaca tcgccgtctg cgtagtccgg cccagtatca agttgttctc 1560

agtgagcgcc tccaagcgct ggtgaatagt ctgtgtgccg gccagagccc acatcaccac 1620

catcaccat 1629

<210>5

<211>294

<212>PRT

<213> Artificial Sequence (Artificial Sequence)

<400>5

His His His His His His Ala Asn Gly Tyr Thr Tyr Glu Asp Tyr Gln

1 5 10 15

Asp Thr Ala Lys Trp Leu Leu Ser His Thr Glu Gln Arg Pro Gln Val

20 25 30

Ala Val Ile Cys Gly Ser Gly Leu Gly Gly Leu Val Asn Lys Leu Thr

35 40 45

Gln Ala Gln Thr Phe Asp Tyr Ser Glu Ile Pro Asn Phe Pro Glu Ser

50 55 60

Thr Val Pro Gly His Ala Gly Arg Leu Val Phe Gly Ile Leu Asn Gly

65 70 75 80

Arg Ala Cys Val Met Met Gln Gly Arg Phe His Met Tyr Glu Gly Tyr

85 90 95

Pro Phe Trp Lys Val Thr Phe Pro Val Arg Val Phe Arg Leu Leu Gly

100 105 110

Val Glu Thr Leu Val Val Thr Asn Ala Ala Gly Gly Leu Asn Pro Asn

115 120 125

Phe Glu Val Gly Asp Ile Met Leu Ile Arg Asp His Ile Asn Leu Pro

130 135 140

Gly Phe Ser Gly Glu Asn Pro Leu Arg Gly Pro Asn Glu Glu Arg Phe

145 150155 160

Gly Val Arg Phe Pro Ala Met Ser Asp Ala Tyr Asp Arg Asp Met Arg

165 170 175

Gln Lys Ala His Ser Thr Trp Lys Gln Met Gly Glu Gln Arg Glu Leu

180 185 190

Gln Glu Gly Thr Tyr Val Met Leu Gly Gly Pro Asn Phe Glu Thr Val

195 200 205

Ala Glu Cys Arg Leu Leu Arg Asn Leu Gly Ala Asp Ala Val Gly Met

210 215 220

Ser Thr Val Pro Glu Val Ile Val Ala Arg His Cys Gly Leu Arg Val

225 230 235 240

Phe Gly Phe Ser Leu Ile Thr Asn Lys Val Ile Met Asp Tyr Glu Ser

245 250 255

Gln Gly Lys Ala Asn His Glu Glu Val Leu Glu Ala Gly Lys Gln Ala

260 265 270

Ala Gln Lys Leu Glu Gln Phe Val Ser Leu Leu Met Ala Ser Ile Pro

275 280 285

Val Ser Gly His Thr Gly

290

<210>6

<211>299

<212>PRT

<213> Artificial Sequence (Artificial Sequence)

<400>6

His His His His His His Lys Leu Asp Val Ile Gly Ile Gly Asn Leu

1 5 10 15

Asn Tyr Asp Ile Ile Phe Thr Leu Glu Arg Phe Pro Glu Phe His Glu

20 25 30

Lys Ile Asn Ala Arg Gly Ala His Phe Gly Leu Gly Gly Ala Ala Ala

35 40 45

Asn Thr Ile Ser Trp Leu Ala His Phe Gly Leu Lys Thr Gly Tyr Ile

50 55 60

Gly Ala Val Gly Asn Asp Asp Val Gly Glu Met His Ile Lys Tyr Phe

65 70 75 80

Gln Gly Ile Gly Val Asp Thr Gly Gly Ile Asp Val Val Glu Glu Pro

85 90 95

Ser Gly Val Ala Val Ala Met Val Ala Gly Asp Asp Lys Arg Ile Val

100 105 110

Lys Tyr Pro Gly Ala Asn Leu Arg Arg Arg Phe Lys Pro Glu Tyr Ala

115 120 125

Ser Arg Ala Lys Phe Leu His Leu Ser Ser Asn Pro Pro Glu Leu Ile

130 135 140

Glu Glu Ala Val Asn Phe Ala Ser Gln Arg Gly Ile Lys Val Ser Leu

145 150155 160

Asp Ile Gly Glu Ala Pro Leu Pro Arg Glu Leu Glu Ser Lys Val Asp

165 170 175

Tyr Leu Met Met Asn Glu Asp Glu Tyr Arg Arg Lys Tyr Gly Ser Leu

180 185 190

Asp Pro Ser Leu Cys Arg Ala Lys Asn Leu Val Val Thr Leu Asn Gly

195 200 205

Gly Gly Ala Leu Val Arg Glu Gly Asp Asn Val Phe Glu Val Arg Gly

210 215 220

Leu Ser Ala Lys Val Val Asp Ser Thr Gly Ala Gly Asp Ser Phe Asp

225 230 235 240

Ala Gly Val Ile Tyr Gly Val Leu Asn Gly Trp Ser Leu Leu Asp Ser

245 250 255

Ala Lys Leu Gly Met Leu Leu Ala Tyr Leu Thr Val Gln Lys Val Gly

260 265 270

Ala Arg Ser Ala Ile Val Pro Leu Glu Glu Val Lys Arg Ile Ala Arg

275 280 285

Glu Val Gly Leu Asp Leu Pro Phe Asn Arg Thr

290 295

<210>7

<211>190

<212>PRT

<213> Artificial Sequence (Artificial Sequence)

<400>7

Met Gly Lys Leu Ile Trp Leu Met Gly Pro Ser Gly Ser Gly Lys Asp

1 5 10 15

Ser Leu Leu Ala Glu Leu Arg Leu Arg Glu Gln Thr Gln Leu Leu Val

20 25 30

Ala His Arg Tyr Ile Thr Arg Asp Ala Ser Ala Gly Ser Glu Asn His

35 40 45

Ile Ala Leu Ser Glu Gln Glu Phe Phe Thr Arg Ala Gly Gln Asn Leu

50 55 60

Leu Ala Leu Ser Trp His Ala Asn Gly Leu Tyr Tyr Gly Val Gly Val

65 70 75 80

Glu Ile Asp Leu Trp Leu His Ala Gly Phe Asp Val Leu Val Asn Gly

85 90 95

Ser Arg Ala His Leu Pro Gln Ala Arg Ala Arg Tyr Gln Ser Ala Leu

100 105 110

Leu Pro Val Cys Leu Gln Val Ser Pro Glu Ile Leu Arg Gln Arg Leu

115 120 125

Glu Asn Arg Gly Arg Glu Asn Ala Ser Glu Ile Asn Ala Arg Leu Ala

130 135 140

Arg Ala Ala Arg Tyr Thr Pro Gln Asp Cys His Thr Leu Asn Asn Asp

145 150 155 160

Gly Ser Leu Arg Gln Ser Val Asp Thr Leu Leu Thr Leu Ile His Gln

165 170 175

Lys Glu Lys His His Ala Cys Leu His His His His His His

180 185 190

<210>8

<211>543

<212>PRT

<213> Artificial Sequence (Artificial Sequence)

<400>8

Ala Ala Glu Gln Asp Pro Glu Ala Arg Ala Ala Ala Arg Pro Leu Leu

1 5 10 15

Thr Asp Leu Tyr Gln Ala Thr Met Ala Leu Gly Tyr Trp Arg Ala Gly

20 25 30

Arg Ala Arg Asp Ala Ala Glu Phe Glu Leu Phe Phe Arg Arg Cys Pro

35 40 45

Phe Gly Gly Ala Phe Ala Leu Ala Ala Gly Leu Arg Asp Cys Val Arg

50 55 60

Phe Leu Arg Ala Phe Arg Leu Arg Asp Ala Asp Val Gln Phe Leu Ala

65 70 75 80

Ser Val Leu Pro Pro Asp Thr Asp Pro Ala Phe Phe Glu His Leu Arg

85 90 95

Ala Leu Asp Cys Ser Glu Val Thr Val Arg Ala Leu Pro Glu Gly Ser

100 105 110

Leu Ala Phe Pro Gly Val Pro Leu Leu Gln Val Ser Gly Pro Leu Leu

115 120 125

Val Val Gln Leu Leu Glu Thr Pro Leu Leu Cys Leu Val Ser Tyr Ala

130 135 140

Ser Leu Val Ala Thr Asn Ala Ala Arg Leu Arg Leu Ile Ala Gly Pro

145 150 155 160

Glu Lys Arg Leu Leu Glu Met Gly Leu Arg Arg Ala Gln Gly Pro Asp

165 170 175

Gly Gly Leu Thr Ala Ser Thr Tyr Ser Tyr Leu Gly Gly Phe Asp Ser

180 185 190

Ser Ser Asn Val Leu Ala Gly Gln Leu Arg Gly Val Pro Val Ala Gly

195 200 205

Thr Leu Ala His Ser Phe Val Thr Ser Phe Ser Gly Ser Glu Val Pro

210 215 220

Pro Asp Pro Met Leu Ala Pro Ala Ala Gly Glu Gly Pro Gly Val Asp

225 230 235 240

Leu Ala Ala Lys Ala Gln Val Trp Leu Glu Gln Val Cys Ala His Leu

245 250 255

Gly Leu Gly Val Gln Glu Pro His Pro Gly Glu Arg Ala Ala Phe Val

260 265 270

Ala Tyr Ala Leu Ala Phe Pro Arg Ala Phe Gln Gly Leu Leu Asp Thr

275 280 285

Tyr Ser Val Trp Arg Ser Gly Leu Pro Asn Phe Leu Ala Val Ala Leu

290 295 300

Ala Leu Gly Glu Leu Gly Tyr Arg Ala Val Gly Val Arg Leu Asp Ser

305 310 315 320

Gly Asp Leu Leu Gln Gln Ala Gln Glu Ile Arg Lys Val Phe Arg Ala

325 330 335

Ala Ala Ala Gln Phe Gln Val Pro Trp Leu Glu Ser Val Leu Ile Val

340 345 350

Val Ser Asn Asn Ile Asp Glu Glu Ala Leu Ala Arg Leu Ala Gln Glu

355 360 365

Gly Ser Glu Val Asn Val Ile Gly Ile Gly Thr Ser Val Val Thr Cys

370 375 380

Pro Gln Gln Pro Ser Leu Gly Gly Val Tyr Lys Leu Val Ala Val Gly

385 390 395 400

Gly Gln Pro Arg Met Lys Leu Thr Glu Asp Pro Glu Lys Gln Thr Leu

405 410 415

Pro Gly Ser Lys Ala Ala Phe Arg Leu Leu Gly Ser Asp Gly Ser Pro

420 425 430

Leu Met Asp Met Leu Gln Leu Ala Glu Glu Pro Val Pro Gln Ala Gly

435 440 445

Gln Glu Leu Arg Val Trp Pro Pro Gly Ala Gln Glu Pro Cys Thr Val

450 455 460

Arg Pro Ala Gln Val Glu Pro Leu Leu Arg Leu Cys Leu Gln Gln Gly

465 470 475 480

Gln Leu Cys Glu Pro Leu Pro Ser Leu Ala Glu Ser Arg Ala Leu Ala

485 490 495

Gln Leu Ser Leu Ser Arg Leu Ser Pro Glu His Arg Arg Leu Arg Ser

500 505 510

Pro Ala Gln Tyr Gln Val Val Leu Ser Glu Arg Leu Gln Ala Leu Val

515 520 525

Asn Ser Leu Cys Ala Gly Gln Ser Pro His His His His His His

530 535 540

Claims (10)

1. A health-care product for pet is prepared from β -1, 3-glucan 0.01-50 wt% and nicotinamide mononucleotide 0.01-50 wt%.

2. The pet health product of claim 1, wherein the mass ratio of β -1,3 glucan to nicotinamide mononucleotide is 1 (1-100).

3. The pet health product of claim 1, wherein said nicotinamide mononucleotide is more than 99% pure and said β -1,3 glucan is more than 70% pure.

4. The pet food of claim 1, further comprising an additional active ingredient comprising at least one of a vitamin, an antioxidant, and a mineral.

5. The pet health product of claim 4, wherein the vitamins include one or more of vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, and vitamin E; the antioxidant comprises one or more of butylated hydroxyanisole, dibutyl hydroxy toluene, propyl gallate, D-sodium erythorbate and tea polyphenol; the mineral comprises mineral elements including one or more of magnesium, iron, zinc, copper, calcium, iodine and manganese.

6. The pet food of claim 1, further comprising one or more of a filler, an excipient, and a preservative.

7. The pet health product of claim 1, wherein the daily amount of β -1,3 glucan in the pet health product is 1-1000 micrograms/kg body weight and the nicotinamide mononucleotide is 1-2000 micrograms/kg body weight.

8. A method of preparing a pet health product according to any one of claims 1 to 7, comprising:

weighing the components according to the formula proportion, putting the components into a stirring pot, and stirring to obtain a mixed material; and then, after forming and drying, cooling to room temperature, and then accommodating in a packaging container and sealing to obtain the pet health product.

9. The method of claim 8, wherein said β -1,3 glucan is produced by a fermentation process comprising:

preparing a culture solution, inoculating zymophyte into the culture solution, fermenting at 28-32 ℃ until the OD600 value of the culture solution is more than 0.6-0.9, and then separating and extracting β -1,3 glucan, wherein the culture solution contains potassium dihydrogen phosphate, sodium nitrate, magnesium sulfate, calcium chloride, ferrous chloride, sucrose and agar.

10. The method of claim 8, wherein the method of synthesizing nicotinamide mononucleotide comprises:

constructing a biological enzyme recombinant plasmid, and performing protein expression and purification to obtain biological enzymes, wherein the biological enzymes comprise a first biological enzyme, a second biological enzyme, a third biological enzyme and a fourth biological enzyme; the first biological enzyme comprises the amino acid sequence as set forth in SEQ ID NO:1 and the second biological enzyme comprises the nucleotide sequence shown as SEQ ID NO: 2, and the third biological enzyme comprises the nucleotide sequence shown as SEQ ID NO: 3 and the fourth biological enzyme comprises the nucleotide sequence shown as SEQ ID NO: 4;

preparing a reaction solution, wherein the reaction solution contains inosine, adenosine diphosphate and adenosine triphosphate, adding the first biological enzyme, the second biological enzyme and the third biological enzyme into the reaction solution, generating 5-phosphoribosyl pyrophosphate after biocatalysis, adding nicotinamide and the fourth biological enzyme, and separating to obtain nicotinamide mononucleotide after biocatalysis.

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