CN103740780B - A kind of method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone - Google Patents
A kind of method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone Download PDFInfo
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Abstract
The invention discloses a kind of method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone, comprise the following steps: after the Lactobacillus plantarum activated culture medium of Lactobacillus plantarum PQQ 1 being stored in agar subculture medium is activated, 20～40 DEG C of quiescent culture 16～30h become first order seed, first order seed continues 20～40 DEG C of expansion quiescent culture 16～30h through primary-seed medium and becomes secondary seed, secondary seed enters yeastiness after secondary seed medium continues 20～40 DEG C of expansion quiescent culture 42～54h, 20～40 DEG C of Anaerobic culturel 54～66h in producing fermentation medium, obtain PQQ.Genus lactubacillus used, in grade-safe microorganism, obtains PQQ by fermentation, and productivity is high, has a good application prospect.
Description
Technical field
The present invention relates to field of microbial fermentation, be specifically related to a kind of by lactic acid bacteria fermentation synthesis pyrroles's quinoline
The method of quinoline quinone.
Background technology
Pyrroloquinoline quinone (pyrroloquinoline quinone, be called for short PQQ) be continue flavin nucleotide and
The third prothetic group found in bacterial dehydrogenase after nicotinamide nucleotide, its chemical name is 4,5-
Dihydro-4,5-titanium dioxide-1-hydrogen pyrroles (2,3f) quinone-2,7,9-tricarboxylic acids, have another name called Methaxatin.Natural conditions
Under only Methylovorus, Deinococcus radiodurans, Gluconobacter oxydans,
The minority antibacterials such as Klebsiella pneumoniae can synthesize (Xiong et al, 2011；Magnusson et
al,2007；Khairnar et al,2003；Felder et al, 2000).
Although only minority antibacterial can synthesize PQQ, but has been found that in various plants, animal and human body
Containing trace P QQ.Japanese Scientists determines the PQQ content in 26 kinds of common food, finds 1
In gram food, PQQ content is at 3.65-61 nanogram, such as, the parsley in vegetable, Capsicum annuum L., fruit
In kiwi fruit, Fructus Chaenomelis, the green tea in beverage, oolong tea, and the bean curd that people often eat.It is worth note
Meaning, in the middle of japanese traditional food natto, PQQ content is the highest, reaches 61 ng/g.This is probably
Because the food that natto is Semen sojae atricolor to be made after antibacterial is fermented for a long time, antibacterial therein constantly secretes PQQ
Create concentration effect.
American Studies personnel detect discovery, and in human milk, PQQ content is up to 140-180 nanograms/milliliter unexpectedly.
Any physiological process has its necessity and rationality.The mankind, through the long-term evolution of millions of years, are developed
Go out the Efficient Operation mechanism of a set of adaptation environment.Containing such high concentration PQQ in breast milk, this thing is described
The growth promoter of confrontation newborn infants may play vital effect.
Since PQQ can only be synthesized by some antibacterial, then the PQQ in plant, animal and human body again from
What？At present, although scientific and technological circle are it is believed that there is the antibacterial of abundant species in animals and humans intestinal,
But all can not synthesize PQQ or synthetic quantity is few, it is impossible to meet somagenic need.Therefore, animals and humans
PQQ can only be obtained by diet approach.And the PQQ in plant is to absorb or oneself from environment
Synthesize, or both have both at the same time, the most indefinite.
PQQ is as a kind of oxidoreductase prothetic group being prevalent in animal and plant tissue, although contain
Measure the lowest, but act on important, be not only a kind of newfound mitochondrial respiratory chain component, participate in catalysis
Organism internal oxidition reduction reaction, also has the most various biological activity, and suckling can be caused during shortage to move
The many dysbolismus of thing it is considered to be a kind of novel vitamin (Misra et al, 2012；Rucker et
al,2012；Kasahara et al,2005；Rucker et al,2005；Toyama et al,2005；Storms et
al,2003；He et al,2003；Bishop et al, 1998).
The major physiological effect of mammal is had by PQQ:
Improve immune function of human body the most comprehensively, as the necessary factor of human developmental, human body can be stimulated thin
Intracellular growth, especially human activin B cell, T cell, improve immune function of human body；
2. preventing and treating hepatic injury, can significantly reduce serum bilirubin and gpt level, liver damages in conditioning
Wound, has fabulous curative effect to hepatic disease；
3. reducing the injury of radical pair human body, PQQ is a kind of oxidoreductase prothetic group, participates in biology
Vivo oxidation reduction reaction, can effectively remove interior free yl, reduces radical pair human injury and is caused
Various diseases, such as heart disease, cancer and all kinds of inflammation；
4. nursing one's health various nervous system disease, PQQ can promote that in human body neural factor synthesizes, conditioning
Various nervous system disease；
5. promoting Amino Acid Absorption, PQQ is the prothetic group of quinoprotein enzyme, participates in respiratory chain electron transmission,
Amino Acid Absorption can be promoted in human body；
6. can promote that somatomedin synthesizes, somatomedin is the hormone of human growth, and human body can be stimulated thin
Intracellular growth, increases cell density, and trace P QQ just can improve metabolic capacity and the growth of bio-tissue
Function；
7. senile dementia prevention and cure, PQQ has reparation nerve fiber, activation neuron, activates dormancy
The ability of neurocyte, can effective senile dementia prevention and cure and improve memory；
8. promoting glutathione synthesis, glutathion has important antioxidation and integrates Detoxication,
PQQ can promote body glutathione synthesis, prevents cataract from occurring and liver bilirubin accumulates；
The most extremely strong anti-cancer function, activating natural killer cell (NK) cell is ANK cell, makes
It has and kills function of tumor；Make the immunocytes such as NK cell assemble, more effectively kill tumor cell；
Close tumor cell and carry Human Placental Ferritin Receptor, block its function, suppress tumor growth, transfer；Destroy tumor
Cellular lipid bilayer, makes oncolysis dead；Stop DNA of tumor cell to replicate, promote its apoptosis.
Research table to the typical bacterium Klebsiella pneumoniae metabolic pathway with synthesis PQQ ability
Bright (Magnusson et al, 2007), synthesizes gene necessary to PQQ and has 6, named
PqqABCDEF, but concrete ways is unclear, only knows that pqqC is catalyzed final step synthesis step, will
3a-(2-amino-2-carboxyethyl)-4,5-dioxo-4,5,6,7,8,9-hexahydroquinoline-7,9-dicar
Boxylic acid (AHQQ) is converted into PQQ.The HPLC isochromatic spectrum method of quantitative determination PQQ is
Set up (Noji N, Nakamura T, Kitahata N, et al.Simple and sensitive method for
pyrroloquinoline quinone(PQQ)analysis in various foods using liquid
chromatography/electrospray-lonization tandem mass spectrometry.J Agri Food
Chem, 2007,55 (18): 7258-7263).
PQQ, as a kind of novel vitamin and powerful abundant bioactie agent, the most still locates
In phase of basic research, the method for there is no implements its large-scale production, and major obstacle is have synthesis PQQ
The microorganism low yield of ability and be all not belonging to grade-safe microorganism, route of synthesis is the most unclear,
And may possess PQQ synthesis plant the most all low yield of potentiality and route of synthesis is known nothing especially.
While it is true, the most still carried out the research of fermentable synthesis PQQ and obtained certain progress.
Xiong etc. (2011) Methylovorus sp.MP688 fermentation synthesis PQQ, productivity reaches 125mg/L
(Xiong XH, Zhao Y, Ge X, et al.Production and Radioprotective Effects of
Pyrroloquinoline Quinone.Inter J Mol Sci, 2011,12 (12): 8913-8923)；Yang etc.
(2010) the pqqABCDE gene cluster of Gluconobacter oxydans is transformed into E.coli so that it is
In fermentation liquid, PQQ concentration reaches 6mM(Yang XP, Zhong GF, Lin JP, et al.
Pyrroloquinoline quinone biosynthesis in Escherichia coli through expression of
the Gluconobacter oxydans pqqABCDE gene cluster.J Indust Microbiol
Biotechnol, 2010,37 (6): 575-580).But that regrettably, have now been found that or recombination to construct has
The antibacterial equal nonfood grade safe microorganisms of PQQ synthesis capability, therefore, for realizing the scale of PQQ
Produce, be badly in need of the grade-safe microorganism that selection-breeding PQQ productivity is high, production performance is good.Lactic acid bacteria is not
Being only generally regarded as safe food-grade microorganisms, and kind is various, genetic resources enriches, breeding high-yield
The lactic acid bacteria of PQQ does not only have abundant feasibility, and is the first-selected approach realizing PQQ large-scale production.
Summary of the invention
The invention provides a kind of method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone, lactic acid bacteria used
For Lactobacillus plantarum Lactobacillus plantarum PQQ-1, belong to grade-safe microorganism, pass through
Fermentation obtains PQQ, and productivity is high.
A kind of method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone, comprises the following steps:
The Lactobacillus plantarum Lactobacillus plantarum of agar subculture medium (A) will be stored in
After the activated culture medium of PQQ-1 (B) activation, 20 DEG C～40 DEG C of quiescent culture 16h～30h become one-level
Seed, first order seed continues 20 DEG C～40 DEG C through primary-seed medium (C1) and expands quiescent culture
16h～30h becomes secondary seed, and secondary seed continues 20 DEG C～40 DEG C through secondary seed medium (C2)
Expand and enter yeastiness after quiescent culture 42h～54h, in producing fermentation medium (D) 20 DEG C～40 DEG C
Anaerobic culturel 54h～66h, obtains PQQ.
Agar subculture medium (A) is used for preserving Lactobacillus plantarum Lactobacillus plantarum
PQQ-1, activation medium (B) is lived for Lactobacillus plantarum Lactobacillus plantarum PQQ-1
Changing propagation, primary-seed medium (C1) and secondary seed medium (C2) are respectively used to plant breast
Bacillus Lactobacillus plantarum PQQ-1 breeds formation high density seed step by step, then accesses life
Produce fermentation medium (D) to start to synthesize PQQ.
In the present invention, Lactobacillus plantarum Lactobacillus plantarum PQQ-1 system is from snow lotus live body (Tibet
Kefir, a kind of symbiosis beneficial flora being made up of lactic acid bacteria, acetic acid bacteria, yeast etc.) in isolated,
Belong to grade-safe microorganism.Concrete separation process is: to snow lotus live body, (Tibet kefir, one is by breast
The symbiosis beneficial flora of the compositions such as acid bacterium, acetic acid bacteria, yeast) add appropriate amounts of sterilized water, by snow lotus live body granule
After pulverizing in milling, dilution, the coating MRS solid plate training containing little mouse-anti PQQ IgG1 monoclonal antibody
Supporting base, quiescent culture 48h～72h, periphery of bacterial colonies forms precipitation circle person, for producing pyrroloquinoline quinone (PQQ)
Bacterium, identified, it is thus achieved that product PQQ bacterium be Lactobacillus plantarum Lactobacillus plantarum, name
For Lactobacillus plantarum PQQ-1.Lactobacillus plantarum Lactobacillus plantarum PQQ-1
It is grade-safe microorganism, commercially available prod can be used, purchased from Yuan Ke bio tech ltd, Jiaxing,
Article number is YK-S-003.Little mouse-anti PQQ IgG1 monoclonal antibody can use commercially available prod, purchased from Jiaxing section of unit
Bio tech ltd, article number is YK-Ab-013.
Further, described agar subculture medium (A) is in terms of 1L, including the component of following weight:
The formula of this agar subculture medium is conducive to Lactobacillus plantarum Lactobacillus plantarum
The successive transfer culture of PQQ-1.
Further, this agar subculture medium acid-base buffer regulates pH to 6.8 so that this agar
Subculture medium has suitable pH, is more suitable for Lactobacillus plantarum Lactobacillus plantarum
The successive transfer culture of PQQ-1.
Described activation medium (B) is in terms of 1L, including the component of following weight:
The formula of this activation medium is conducive to Lactobacillus plantarum Lactobacillus plantarum PQQ-1
Activation culture.
Further, this activation medium acid-base buffer regulates pH to 6.8 so that this activation culture
Base has suitable pH, is more suitable for the work of Lactobacillus plantarum Lactobacillus plantarum PQQ-1
Change and cultivate.
Further, 30 DEG C of quiescent culture 24h become first order seed.
Described primary-seed medium (C1) is in terms of 1L, including the component of following weight:
The formula of this primary-seed medium is conducive to Lactobacillus plantarum Lactobacillus plantarum
The cultivation of PQQ-1.
Further, this primary-seed medium acid-base buffer regulates pH to 6.8 so that this one-level
Seed culture medium has suitable pH, is more suitable for Lactobacillus plantarum Lactobacillus plantarum
The cultivation of PQQ-1.
Further, first order seed continues 30 DEG C through primary-seed medium (C1) and expands 50 times of standing trainings
Foster 24h becomes secondary seed.
Further, described secondary seed medium (C2) is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus.
The formula of this secondary seed medium is conducive to Lactobacillus plantarum Lactobacillus plantarum
The amplification culture of PQQ-1.
Further, this secondary seed medium acid-base buffer regulates pH to 6.8 so that these two grades
Seed culture medium has suitable pH, is more suitable for Lactobacillus plantarum Lactobacillus plantarum
The amplification culture of PQQ-1.
Further, secondary seed continues 30 DEG C through secondary seed medium (C2) and expands 50 times of standing trainings
Yeastiness is entered after supporting 48h.
Further, described production fermentation medium (D) is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus.
The formula of this production fermentation medium is conducive to Lactobacillus plantarum Lactobacillus plantarum
The production fermentation of PQQ-1.
Further, this production fermentation medium acid-base buffer regulates pH to 6.8 so that this production
Fermentation medium has suitable pH, is more suitable for Lactobacillus plantarum Lactobacillus plantarum
The production fermentation of PQQ-1.
Further, in producing fermentation medium (D), 30 DEG C expand 50 times of Anaerobic culturel 60h, obtain
PQQ。
Compared with prior art, present invention have the advantage that
One, the present invention selects Lactobacillus plantarum Lactobacillus plantarum PQQ-1, is from snow lotus live body
(Tibet kefir, a kind of symbiosis beneficial flora being made up of lactic acid bacteria, acetic acid bacteria, yeast etc.) separates
Obtain, belong to grade-safe microorganism.
Two, this Lactobacillus plantarum Lactobacillus plantarum PQQ-1 fermentation synthesis PQQ's is final
Productivity is high, may be up to 238ng/mL, the human milk the highest far above content in nature
(140-180ng/mL), natto (55-63ng/mL) and bean curd (25-30ng/mL), the most significantly high
In the Methylovorus sp.MP688(Xiong XH having now been found that and building, Zhao Y, Ge X, et al.
Production and Radioprotective Effects of Pyrroloquinoline Quinone.Inter J Mol
And introduce the E.coli(Yang XP of pqqABCDE gene cluster, Sci, 2011,12 (12): 8913-8923.)
Zhong GF,Lin JP,et al.Pyrroloquinoline quinone biosynthesis in Escherichia
coli through expression of the Gluconobacter oxydans pqqABCDE gene cluster.J
Indust Microbiol Biotechnol, 2010,37 (6): 575-580.).
Accompanying drawing explanation
Fig. 1 is the schematic flow sheet that the present invention passes through the method for lactic acid bacteria fermentation synthesis pyrroloquinoline quinone.
Detailed description of the invention
Embodiment 1
As it is shown in figure 1, the present invention is by the method for lactic acid bacteria fermentation synthesis pyrroloquinoline quinone, including following
Step:
(1) Lactobacillus plantarum PQQ-1 uses commercially available prod, biological purchased from Jiaxing section of unit
Science and Technology Ltd., article number is YK-S-003.
(2) each culture medium, wherein, yeast extract and albumen are prepared by the component of following content respectively
Peptone is Sigma product, and Fructus Hordei Vulgaris powder is Dafeng City of Jiangsu Province gold wheat prosperous product of edge wheat core；Other reagent is adopted
Use analytical pure；After having prepared, all at 121 DEG C of sterilizing 15min；After having prepared, all at 121 DEG C
Sterilizing 15min；
(a): MRS solid plate culture medium is in terms of 1L, including the component of following weight:
With sulphuric acid, pH value is adjusted to 6.5 ± 0.2.
(b): agar subculture medium (A) is in terms of 1L, including the component of following weight:
And this agar subculture medium acid-base buffer is regulated pH to 6.8.
(c): activation medium (B) is in terms of 1L, including the component of following weight:
And this activation medium acid-base buffer is regulated pH to 6.8.
(d): primary-seed medium (C1) is in terms of 1L, including the component of following weight:
And this primary-seed medium acid-base buffer is regulated pH to 6.8.
(e): secondary seed medium (C2) is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus；
And this secondary seed medium acid-base buffer is regulated pH to 6.8.
(f): production fermentation medium (D) is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus；
And this production fermentation medium acid-base buffer regulates pH to 6.8.
(3) bacterial strain activation: 10 milliliters of activation mediums (B) are poured in 30 milliliters of test tubes, at test tube
Middle inoculation one ring freezing test tube strains Lactobacillus plantarum Lactobacillus plantarum PQQ-1,30 DEG C quiet
Put cultivation 24h and become first order seed；
(4) first order seed is cultivated: takes 10mL first order seed and accesses 500mL primary-seed medium
(C1), in, 30 DEG C of quiescent culture 24h become secondary seed；
(5) secondary seed is cultivated: takes 100mL secondary seed and accesses 5000mL secondary seed medium
(C2), in, yeastiness after 30 DEG C of quiescent culture 48h, is entered；
(6) PQQ fermentation: take 1000mL and enter the secondary seed access 50L production of yeastiness
In ferment culture medium (D), 30 DEG C stand Anaerobic culturel 60h, obtain PQQ.
Use chromatography determination PQQ, produce PQQ concentration in fermentation medium (D) and reach 238ng/mL.
This chromatography determination PQQ method concrete steps are with reference to " Noji N, Nakamura T, Kitahata N, et al.
Simple and sensitive method for pyrroloquinoline quinone(PQQ)analysis in
various foods using liquid chromatography/electrospray-lonization tandem mass
spectrometry.J Agri Food Chem,2007,55(18):7258-7263.”。
Embodiment 2
As it is shown in figure 1, the present invention is by the method for lactic acid bacteria fermentation synthesis pyrroloquinoline quinone, including following
Step:
(1) prepare each culture medium by the component of following content respectively, after having prepared, all go out at 121 DEG C
Bacterium 15min, yeast extract and peptone are Sigma product, and Fructus Hordei Vulgaris powder is Dafeng City of Jiangsu Province gold
The prosperous product of wheat edge wheat core；Other reagent uses analytical pure；After having prepared, all at 121 DEG C of sterilizing 15min；
(a): agar subculture medium (A) is in terms of 1L, including the component of following weight:
And this agar subculture medium acid-base buffer is regulated pH to 6.8.
(b): activation medium (B) is in terms of 1L, including the component of following weight:
And this activation medium acid-base buffer is regulated pH to 6.8.
(c): primary-seed medium (C1) is in terms of 1L, including the component of following weight:
And this primary-seed medium acid-base buffer is regulated pH to 6.8.
(d): secondary seed medium (C2) is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus；
And this secondary seed medium acid-base buffer is regulated pH to 6.8.
(e): production fermentation medium (D) is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus；
And this production fermentation medium acid-base buffer regulates pH to 6.8.
(2) bacterial strain activation: 10 milliliters of activation mediums (B) are poured in 30 milliliters of test tubes, at test tube
Middle inoculation one ring freezing test tube strains Lactobacillus plantarum Lactobacillus plantarum PQQ-1(Jiaxing unit
Bio tech ltd of section, article number is YK-S-003), 30 DEG C of quiescent culture 24h become one-level kind
Son；
(3) first order seed is cultivated: takes 10mL first order seed and accesses 500mL primary-seed medium
(C1), in, 30 DEG C of quiescent culture 24h become secondary seed；
(4) secondary seed is cultivated: takes 100mL secondary seed and accesses 5000mL secondary seed medium
(C2), in, yeastiness after 30 DEG C of quiescent culture 48h, is entered；
(5) PQQ fermentation: take 5000mL and enter the secondary seed access 50L production of yeastiness
In ferment culture medium (D), 30 DEG C stand Anaerobic culturel 60h, obtain PQQ.
Use chromatography determination PQQ, produce PQQ concentration in fermentation medium (D) and reach 229ng/mL,
Measure PQQ method with embodiment 1.
Claims (7)
1. one kind by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone method, it is characterised in that include with
Lower step:
Lactic acid bacteria uses Lactobacillus plantarum Lactobacillus plantarum PQQ-1, will be stored in agar and continue
After the Lactobacillus plantarum Lactobacillus plantarum PQQ-1 activated culture medium activation of culture base,
20 DEG C～40 DEG C of quiescent culture 16h～30h become first order seed, and first order seed continues through primary-seed medium
Continuous 20 DEG C～40 DEG C expansion quiescent culture 16h～30h become secondary seed, and secondary seed is trained through secondary seed
Support after base continues 20 DEG C～40 DEG C expansion quiescent culture 42h～54h and enter yeastiness, producing fermentation training
Support in base 20 DEG C～40 DEG C of Anaerobic culturel 54h～66h, obtain pyrroloquinoline quinone；
Described primary-seed medium is in terms of 1L, including the component of following weight:
Described secondary seed medium is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus；
Described production fermentation medium is in terms of 1L, including the component of following weight:
Wholegrain Fructus Hordei Vulgaris powder 30g；
Yeast extract 7.5g；
Water surplus.
Method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone the most according to claim 1, its
Being characterised by, described agar subculture medium is in terms of 1L, including the component of following weight:
Method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone the most according to claim 2, its
Being characterised by, this agar subculture medium acid-base buffer regulates pH to 6.8.
Method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone the most according to claim 1, its
Being characterised by, described activation medium is in terms of 1L, including the component of following weight:
Method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone the most according to claim 4, its
Being characterised by, this activation medium acid-base buffer regulates pH to 6.8.
Method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone the most according to claim 1, its
It is characterised by, described primary-seed medium acid-base buffer regulation pH to 6.8.
Method by lactic acid bacteria fermentation synthesis pyrroloquinoline quinone the most according to claim 1, its
It is characterised by, described secondary seed medium acid-base buffer regulation pH to 6.8.
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| CN104522182A (en) * | 2014-12-10 | 2015-04-22 | 浙江大学 | Method for preparing fermented mung bean milk containing dietary therapy components |
| CN104489116A (en) * | 2014-12-10 | 2015-04-08 | 浙江大学 | Method for preparing peanut milk through co-fermentation of selected bacillus natto and lactobacillus casei |
| US11083762B2 (en) | 2017-12-18 | 2021-08-10 | Industrial Technology Research Institute | Application for water extract of plant of Gracilariaceae or ferment thereof and pharmaceutical composition and/or health food for treating and/or alleviating nervous diseases |
| CN108949846A (en) * | 2018-08-01 | 2018-12-07 | 安徽新熙盟生物科技有限公司 | A kind of method that high density fermentation improves PQQ yield |
| CN112602926B (en) * | 2020-12-30 | 2022-08-19 | 浙江大学 | Plant enzyme rich in pyrroloquinoline quinone and gamma-aminobutyric acid and preparation method thereof |
| CN112741241B (en) * | 2020-12-30 | 2022-08-16 | 浙江大学 | Chrysanthemum morifolium fermented beverage rich in pyrroloquinoline quinone and preparation method thereof |
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