CN113789276B - Lactobacillus plantarum for producing beta-nicotinamide mononucleotide and application thereof - Google Patents
Lactobacillus plantarum for producing beta-nicotinamide mononucleotide and application thereof Download PDFInfo
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- CN113789276B CN113789276B CN202111081824.2A CN202111081824A CN113789276B CN 113789276 B CN113789276 B CN 113789276B CN 202111081824 A CN202111081824 A CN 202111081824A CN 113789276 B CN113789276 B CN 113789276B
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
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Abstract
The invention discloses a lactobacillus plantarum for producing beta-nicotinamide mononucleotide and application thereof, belonging to the technical field of microbial fermentation, wherein the lactobacillus plantarum is lactobacillus plantarum (Lactobacillus plantarum) QCT009, the registration number of the lactobacillus plantarum in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No.21881, and the lactobacillus plantarum is applied to the preparation of the beta-nicotinamide mononucleotide. The invention also discloses a method for preparing beta-nicotinamide mononucleotide by using lactobacillus plantarum QCT 009. Compared with the existing microbial fermentation method for preparing the beta-nicotinamide mononucleotide, the lactobacillus plantarum QCT009 provided by the invention has higher yield, and solves the problem of low yield of the existing microbial fermentation method for preparing the beta-nicotinamide mononucleotide.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to lactobacillus plantarum for producing beta-nicotinamide mononucleotide and application thereof.
Background
Beta-nicotinamide mononucleotide (Nicotinamide mononucleotide): NMN is an important substance in human body cell energy generation, participates in synthesis of nicotinamide adenine dinucleotide (NAD+) in cells, can effectively prevent and treat various diseases caused by aging by taking NMN as a precursor of NAD+, and has medical care effects in aspects of delaying aging, treating senile diseases such as Parkinson, regulating insulin secretion, regulating mRNA expression and the like. The artificial NMN supplement can repair brain injury, improve islet function, protect heart from ischemia reperfusion injury, repair brain mitochondrial respiratory defect, and has certain treatment effects on senile degenerative diseases, retinal degenerative diseases and the like. But is limited by the synthesis process at present, NMN raw materials are expensive, and the application of NMN as medicines is limited. At present, NMN is prepared mainly by a chemical synthesis method, a microbial fermentation method and an in-vitro enzyme conversion method. Among them, chemical synthesis techniques are most mature, but are costly and present problems of chiral compounds and food safety; the in vitro enzyme conversion method needs various substrates of enzyme conversion reaction, and the cost of the substrates is high; the microbial fermentation method only needs nicotinamide as a reaction substrate, and has low cost, but the yield of products obtained by the currently used escherichia coli fermentation method and yeast fermentation method is low, and the requirements of commercial production cannot be met.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: a strain for preparing a beta-nicotinamide mononucleotide and a method for preparing a beta-nicotinamide mononucleotide using the strain are provided.
In order to solve the technical problems, the invention adopts the following technical scheme: a lactobacillus plantarum for producing beta-nicotinamide mononucleotide is lactobacillus plantarum (Lactobacillus plantarum) QCT009, and the registration number of the lactobacillus plantarum in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC NO.21881.
The lactobacillus plantarum is applied to the preparation of beta-nicotinamide mononucleotide.
A beta-nicotinamide mononucleotide prepared from Lactobacillus plantarum QCT009
A method for preparing beta-nicotinamide mononucleotide, comprising the following steps:
activating the lactobacillus plantarum QCT009, inoculating the activated lactobacillus plantarum into a beta-nicotinamide mononucleotide production medium for culture, and obtaining a culture solution; and (3) separating and purifying the obtained culture solution after cell disruption to obtain the beta-nicotinamide mononucleotide.
The invention has the beneficial effects that: the lactobacillus plantarum QCT009 provided by the invention can be used for preparing beta-nicotinamide mononucleotide, and compared with the existing microbial fermentation for preparing beta-nicotinamide mononucleotide, such as escherichia coli fermentation and yeast fermentation, the yield is higher, and the problem of low yield of the beta-nicotinamide mononucleotide prepared by the existing microbial fermentation is solved.
Detailed Description
In order to describe the technical contents, the achieved objects and effects of the present invention in detail, the following description will be made with reference to the embodiments.
The most critical concept of the invention is as follows: and screening the natto surface to obtain the strain for producing the beta-nicotinamide mononucleotide.
The invention provides a lactobacillus plantarum for producing beta-nicotinamide mononucleotide, which is obtained by culturing the following method:
selecting natto with good quality, adding proper amount of sterile physiological saline, repeatedly stirring in sterile environment with sterile spoon (or sterile stirring rod) to suspend microorganism on natto surface in sterile physiological saline, then taking 0.2mL physiological saline with microorganism suspended in sterile solid plate culture medium, spreading uniformly, culturing in 28 deg.C incubator for 2-5 days until single colony grows on the surface of culture dish, selecting single colony, culturing in liquid culture medium, measuring NMN content in culture medium after culturing, and selecting optimal strain. The culture conditions are as follows: temperature: 28-30 ℃; rotational speed: 0-50rpm; culturing time: 72-240h.
The solid plate culture medium and the liquid culture medium comprise 2-5 parts by weight of agaricus blazei murill extract, 2-10 parts by weight of hericium erinaceus extract, 1-5 parts by weight of yeast powder, 0.2-5 parts by weight of fructo-oligosaccharide and 0.1-2 parts by weight of nicotinamide.
The strain QCT009 is stored in: china general microbiological culture Collection center (CGMCC); address: beijing, chaoyang area, north Chenxi Lu No. 1, 3; the preservation date is: 2021, 03, 09; name: lactobacillus plantarum; latin name: lactobacillus plantarum; strain name: QCT009; preservation number: CGMCC No.21881.
Application of lactobacillus plantarum QCT009 in preparing beta-nicotinamide mononucleotide.
A beta-nicotinamide mononucleotide prepared from Lactobacillus plantarum QCT009 is provided.
A method for preparing beta-nicotinamide mononucleotide, comprising the following steps:
activating lactobacillus plantarum QCT009, inoculating the activated lactobacillus plantarum into a beta-nicotinamide mononucleotide production medium for culture, and obtaining a culture solution; and (3) separating and purifying the obtained culture solution after cell disruption to obtain the beta-nicotinamide mononucleotide.
From the above description, the beneficial effects of the invention are as follows: the lactobacillus plantarum QCT009 provided by the invention can be used for preparing beta-nicotinamide mononucleotide, and compared with the existing microbial fermentation for preparing beta-nicotinamide mononucleotide, such as escherichia coli fermentation and yeast fermentation, the yield is higher, and the problem of low yield of the beta-nicotinamide mononucleotide prepared by the existing microbial fermentation is solved. Provides possibility for the industrialized production of the beta-nicotinamide mononucleotide.
Further, the inoculation amount of the lactobacillus plantarum is 1-5%.
Further, the culture conditions are as follows: temperature: 28-30 ℃; rotational speed: 0-50rpm; culturing time: 72-240h.
Further, the beta-nicotinamide mononucleotide production medium comprises 2-5 parts by weight of agaricus blazei murill extract, 2-10 parts by weight of hericium erinaceus extract, 1-5 parts by weight of yeast powder, 0.2-5 parts by weight of fructo-oligosaccharide and 0.1-2 parts by weight of nicotinamide.
From the above description, it is clear that by optimizing the conditions of lactobacillus plantarum QCT009 fermentation and after the culture medium, a high yield of β -nicotinamide mononucleotide is ensured.
Example 1:
a method for preparing beta-nicotinamide mononucleotide, comprising the following steps:
activating lactobacillus plantarum QCT009, inoculating the activated activating solution into 200mL of beta-nicotinamide mononucleotide production medium according to an inoculum size of 1% for culture to obtain a culture solution;
the culture conditions are as follows: temperature: 28 ℃; rotational speed: 30rpm; culturing time: 120h;
the beta-nicotinamide mononucleotide production medium consists of 5g of agaricus blazei murill extract, 5g of hericium erinaceus extract, 3g of yeast powder, 3g of fructo-oligosaccharide, 1g of nicotinamide and the balance of sterile water;
and (3) separating and purifying the obtained culture solution after cell disruption to obtain the beta-nicotinamide mononucleotide.
Example 2:
a method for preparing beta-nicotinamide mononucleotide, comprising the following steps:
activating lactobacillus plantarum QCT009, inoculating the activated activating solution into 200mL of beta-nicotinamide mononucleotide production medium according to an inoculum size of 1% for culture to obtain a culture solution;
the culture conditions are as follows: temperature: 28 ℃; rotational speed: 50rpm; culturing time: 72h;
the beta-nicotinamide mononucleotide production medium consists of 4g of agaricus blazei murill extract, 10g of hericium erinaceus extract, 5g of yeast powder, 5g of fructo-oligosaccharide, 2g of nicotinamide and the balance of sterile water;
and (3) separating and purifying the obtained culture solution after cell disruption to obtain the beta-nicotinamide mononucleotide.
Example 3:
a method for preparing beta-nicotinamide mononucleotide, comprising the following steps:
activating lactobacillus plantarum QCT009, inoculating the activated activating solution into 200mL of beta-nicotinamide mononucleotide production medium according to an inoculum size of 1% for culture to obtain a culture solution;
the culture conditions are as follows: temperature: 28 ℃; rotational speed: 0rpm; culturing time: 240h;
the beta-nicotinamide mononucleotide production medium consists of 2g of agaricus blazei murill extract, 2g of hericium erinaceus extract, 1g of yeast powder, 0.2g of fructo-oligosaccharide, 0.1g of nicotinamide and the balance of sterile water;
and (3) separating and purifying the obtained culture solution after cell disruption to obtain the beta-nicotinamide mononucleotide.
In summary, lactobacillus plantarum QCT009 provided by the invention can be used for preparing beta-nicotinamide mononucleotide, and compared with the existing microbial fermentation for preparing beta-nicotinamide mononucleotide, such as escherichia coli fermentation and yeast fermentation, the yield is higher, and the problem of low yield of the existing microbial fermentation for preparing beta-nicotinamide mononucleotide is solved.
The foregoing description is only illustrative of the present invention and is not intended to limit the scope of the invention, and all equivalent modifications made by the teachings of the present invention, or direct or indirect application in the relevant art, are intended to be included within the scope of the present invention.
Claims (3)
1. A lactobacillus plantarum producing beta-nicotinamide mononucleotide is characterized in that,
the lactobacillus plantarum is lactobacillus plantarum (Lactobacillus plantarum) QCT009, and the registration number of the lactobacillus plantarum in the common microorganism center of the China Committee for culture Collection of microorganisms is CGMCC NO.21881.
2. Use of lactobacillus plantarum according to claim 1 for the preparation of β -nicotinamide mononucleotide.
3. A method for preparing beta-nicotinamide mononucleotide, which is characterized by comprising the following steps:
activating the lactobacillus plantarum of claim 1, inoculating the activated lactobacillus plantarum into a beta-nicotinamide mononucleotide production medium for culture to obtain a culture solution; the obtained culture solution is subjected to cell disruption and then is separated and purified to obtain beta-nicotinamide mononucleotide; the inoculation amount of the lactobacillus plantarum is 1-5%; the culture conditions are as follows: temperature: 28 ℃; rotational speed: 0-50rpm; culturing time: 72-240h; the beta-nicotinamide mononucleotide production medium comprises 2-5 parts by weight of agaricus blazei murill extract, 2-10 parts by weight of hericium erinaceus extract, 1-5 parts by weight of yeast powder, 0.2-5 parts by weight of fructo-oligosaccharide and 0.1-2 parts by weight of nicotinamide.
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