CN109628350A - Lactobacillus plantarum and its application - Google Patents
Lactobacillus plantarum and its application Download PDFInfo
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- CN109628350A CN109628350A CN201910016017.9A CN201910016017A CN109628350A CN 109628350 A CN109628350 A CN 109628350A CN 201910016017 A CN201910016017 A CN 201910016017A CN 109628350 A CN109628350 A CN 109628350A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
Abstract
The present invention provides a kind of lactobacillus plantarum and its application.Lactobacillus plantarum (Lactobacillus plantarum) of the invention, deposit number are CGMCC No.14812.Lactobacillus plantarum provided by the invention can be used in improving organic acid in raw material, can be fermented using the lactobacillus plantarum to raw material, and wherein raw material can be selected from least one of fruit.It is fermented using lactobacillus plantarum of the invention to raw material, can be improved the content of organic acid in fermented product.
Description
Technical field
The present invention relates to a kind of lactic acid bacterias, and in particular to a kind of lactobacillus plantarum and its application.
Background technique
Organic acid is the acids in molecule containing carboxyl, is widely present in animal, plant and microbial body, mainly includes
Lactic acid, acetic acid, propionic acid, butyric acid, malic acid, tartaric acid, oxalic acid, citric acid etc..Many organic acids can directly participate in bioid
Reaction is learned, some of them organic acid has the effects that antibacterial, anti-inflammatory, antiviral, anti-mutation, anticancer；Some can increase coronal
Blood flow volume inhibits cerebrum lipid peroxide to generate, softening blood vessel, the absorption for promoting calcium, ferro element；Also some have
It helps Gastric juice digestion fat and protein, there is prevention disease and the effect that boosts metabolism, to be beneficial to the health of human body.
Lactic acid bacteria (lactic acid bacteria, LAB) is that one kind can be generated greatly using fermentable carbohydrate
Measure the common name of the bacterium of lactic acid.A large number of studies show that lactic acid bacteria can adjust body gastrointestinal tract normal flora, keep Tiny ecosystem flat
Weighing apparatus improves food digestion rate and biological value, reduces serum cholesterol, control endotoxin, inhibits spoilage organisms growth and breeding in enteron aisle
Generation, manufacture nutriment with spoilage product, stimulation tissue development, thus to the nutritional status of body, physiological function, cell
The generations effects such as infection, drug effect, toxic reaction, immune response, tumour generation, aging course and unexpected emergency reaction.
Lactic acid bacteria generates the substances such as organic acid by fermentation to play its special physiological function.Lactic acid bacteria can will be newborn
Sugar decomposition is the small molecule compounds such as lactic acid at glucose and galactolipin, the fermented role transformation of glucose, facilitates children's brain
The development of tissue and nervous system.Lactic acid bacteria has Phospoprotein enzyme, and alpha-casein can be resolved into fine cheese fat peptide
With amino acid etc., to improve the digestibility of albumen.Lactobacillus-fermented can make partial fat degrade on a small quantity, easy to digest
And it can increase free fatty acid in cream, Volatile fatty acid contents.Lactic acid bacteria consumes some vitamin in the metabolic process, simultaneously
Also the B family vitamins such as folic acid are synthesized.The utilization rate of the elements such as calcium, phosphorus, iron can be improved in the organic acid generated after lactobacillus-fermented, promotees
Into the absorption of iron and VD, and there is stronger bacteriostasis.
Currently, content of the organic acid in plant and microorganism is relatively low, it is therefore contemplated that one kind can be improved organic acid
The method of content.
Summary of the invention
The present invention provides a kind of lactobacillus plantarum and its application, is fermented using the lactobacillus plantarum to raw material, can
Improve the content of organic acid in fermented product.
The present invention provides a kind of lactobacillus plantarum (Lactobacillus plantarum), deposit number CGMCC
No.14812。
The phenotypic characteristic of lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812) of the invention
Are as follows: G+, rod-shaped, bacterium colony is round, milky, neat in edge.
Lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812) of the invention is independently to pass through
Can cultural method separation screening from spontaneous fermentation pectase obtain, utilize 16S rRNA gene order and pheS genic system
Auxology analysis, has determined taxonomy in conjunction with phenotypic characteristic.
The growth conditions of lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812) of the invention
It can be normal condition, growth conditions for example can be with are as follows: 30-37 DEG C of growth temperature, amphimicrobian, growth time 24-48h.
Lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812) of the invention can ferment life
Organic acid is produced, therefore can be used for associated uses.
The present invention also provides above-mentioned lactobacillus plantarums (Lactobacillus plantarum) (CGMCC No.14812) to exist
Produce the application on organic acid.
The present invention is producing above-mentioned lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812)
Application mode on organic acid does not limit strictly, can both produce the fermented product comprising organic acid, can also be as needed
Organic acid is extracted from the fermented product, the application in downstream is provided.
The present invention also provides a kind of fermented product production methods, using above-mentioned lactobacillus plantarum (Lactobacillus
Plantarum) (CGMCC No.14812) ferments to raw material.
The present invention does not limit the raw material strictly, as long as being conducive to above-mentioned lactobacillus plantarum (Lactobacillus
Plantarum) (CGMCC No.14812) produces organic acid.
Specifically, the raw material can be selected from least one of fruit and integration of drinking and medicinal herbs class raw material.Further, described
Fruit includes orange, loquat, dragon fruit and mulberry fruit etc., and the integration of drinking and medicinal herbs class raw material includes Poria cocos, Radix Glycyrrhizae, dark plum and chrysanthemum
Deng.
Using above-mentioned lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812) to raw material into
When row fermentation, fermentation medium and fermentation condition are not limited strictly, as long as being conducive to above-mentioned lactobacillus plantarum
(Lactobacillus plantarum) (CGMCC No.14812) produces organic acid.
In concrete scheme of the invention, fermented product production method be may include steps of:
It squeezed the juice, sized mixing to the raw material, obtain raw material juice；
0-80g carbon source, 3-8g nitrogen source, 1-3g inorganic salts and 5-12g calcium carbonate are added into raw material juice described in every liter, then
PH value is adjusted to 5.5-6.8, obtains fermentation medium；
The lactobacillus plantarum (Lactobacillus plantarum) is seeded to the hair by 0.5-1.5% (v/v)
It ferments in ferment culture medium, obtains fermented product.
In the present invention, usual manner can be used to squeeze the juice to raw material, to obtain raw material Normal juice；In addition, visual real
Border needs to size mixing to raw material Normal juice.Specifically, size mixing can using distilled water carry out, wherein can control raw material Normal juice with
The volume ratio of distilled water is 1:(1-5), so that raw material juice be made.
The present invention does not limit carbon source, nitrogen source, inorganic salts strictly, as long as being suitable for above-mentioned lactobacillus plantarum
(Lactobacillus plantarum) produces organic acid.Specifically, carbon source can use sucrose, glucose, maltose
At least one of Deng, nitrogen source can use at least one of peptone, beef extract, ammonium chloride, ammonium nitrate, potassium nitrate etc.,
Inorganic salts can be using at least one of manganese sulfate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate etc..
Further, calcium carbonate can also be added into raw material juice.With the progress of fermentation, strain not stopping pregnancy acid can be reduced
PH, it is possible to which the growth fermentation of strain is had an impact；So can according to need by the way that calcium carbonate is added, consumption H is played+、
Stablize the effect of pH.
Identical as conventional zymotechnique, the present invention uses above-mentioned short lactobacillus plantarum (Lactobacillus
Plantarum) production organic acid process also includes actication of culture and preparation seed liquor operation, and seed liquor is seeded to hair
Ferment culture medium implements the operation of fermentation.The present invention does not limit fermentation condition strictly, as long as being suitable for above-mentioned lactobacillus plantarum
(Lactobacillus plantarum) produces organic acid.Specifically, the temperature of the fermentation can be 20-35 DEG C, turn
Speed can be 60-150r/min, and fermentation period can be 7-30 days.The studies have shown that of inventor utilizes provided by the invention short
Lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812), compared with similar strain, such as plant cream bar
Bacterium (Lactobacillus plantarum) reference culture ATCC 14917, not only the organic acid content in tunning improves,
And the inoculum concentration needed can reduce, and apparent advantage is all shown in production efficiency and cost.
The present invention also provides a kind of fermented products, produce to obtain according to any of the above-described fermented product production method.
It include organic acid in fermented product of the invention.Further, the organic acid includes lactic acid, acetic acid, propionic acid, fourth
At least one of acid, malic acid, tartaric acid, oxalic acid and citric acid etc..
Lactobacillus plantarum (Lactobacillus plantarum) of the present invention is protected on October 16th, 2017
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC), address is Beijing's southern exposure
The institute 3 of area North Star West Road 1, deposit number are CGMCC No.14812.
Lactobacillus plantarum (Lactobacillus plantarum) (CGMCC No.14812) provided by the invention can be sent out
Ferment produces organic acid, when fermenting using the lactobacillus plantarum to raw material, can be improved the content of organic acid in fermented product.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with the embodiment of the present invention, to this
Technical solution in inventive embodiments is clearly and completely described, it is clear that described embodiment is that a part of the invention is real
Example is applied, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art are not making creation
Property labour under the premise of every other embodiment obtained, shall fall within the protection scope of the present invention.
Embodiment 1
One, culture medium is prepared
1) basal medium
By 10.0g casein peptone, 10.0g beef extract, 5.0g yeast powder, 5.0g glucose, 5.0g sodium acetate, 2.0g lemon
Sour diammonium, 1.0g Tween 80,2.0g K2HPO4、0.2g MgSO4·7H2O、0.05g MnSO4·H2O、10.0g CaCO3Dissolution
In 1L distilled water, pH value is adjusted to 6.8, basal medium (when preparing solid medium, 20g agar is added) is made.
2) fermentation medium
It squeezes the juice to orange peel, obtains orange peel Normal juice；Distilled water is added into orange peel Normal juice to size mixing, control orange peel is former
The volume ratio of juice and distilled water is 1:1, and syrup of organge is made.
50g glucose, 6g peptone, 1.2g manganese sulfate and 10.0g CaCO are added into the above-mentioned syrup of organge of 1L3, adjust pH
Value obtains fermentation medium to 6.8 or so.
Two, it ferments
1) actication of culture
By the lactobacillus plantarum being stored on inclined-plane (Lactobacillus plantarum) strain (CGMCC
No.14812 it) is inoculated in above-mentioned total amount of binder culture medium, stationary culture controls for 24 hours under anaerobic, obtains activated spawn.
2) seed liquor is prepared
The strain of above-mentioned activation is inoculated in aforesaid liquid basal medium, stationary culture is left for 24 hours under anaerobic
The right side obtains seed liquor.
3) it ferments
According to the inoculum concentration of 1% (v/v), above-mentioned seed liquor is inoculated in above-mentioned fermentation medium and carries out fermented and cultured,
Controlling fermentation temperature is 35 DEG C or so, and revolving speed is 150r/min or so, and fermentation period is 7 days or so, obtains orange peel fermented product.
Using People's Republic of China's agricultural industry criteria NY/T 2277-2012 method in above-mentioned orange peel fermented product
Organic acid detected, the results are shown in Table 1.
Embodiment 2
One, culture medium is prepared
1) basal medium
By 8.0g casein peptone, 12.0g beef extract, 4.0g yeast powder, 6.0g glucose, 6.0g sodium acetate, 1.0g lemon
Sour diammonium, 1.5g Tween 80,3.0g K2HPO4、0.1g MgSO4·7H2O、0.1g MnSO4·H2O、8.0g CaCO3It is dissolved in
In 1L distilled water, pH value is adjusted to 6.0, basal medium (when preparing solid medium, 20g agar is added) is made.
2) fermentation medium
It squeezes the juice to loquat, obtains loquat Normal juice；Distilled water is added into loquat Normal juice to size mixing, control loquat is former
The volume ratio of juice and distilled water is 1:2, and loquat juice is made.
60g glucose, 3g peptone, 2.0g manganese sulfate and 8.0g CaCO are added into the above-mentioned loquat juice of 1L3, adjust pH
Value obtains fermentation medium to 6.0 or so.
Two, it ferments
1) actication of culture
By the lactobacillus plantarum being stored on inclined-plane (Lactobacillus plantarum) strain (CGMCC
No.14812 it) is inoculated in above-mentioned total amount of binder culture medium, stationary culture 18h or so, obtains activated spawn under anaerobic.
2) seed liquor is prepared
The strain of above-mentioned activation is inoculated in aforesaid liquid basal medium, stationary culture is left for 24 hours under anaerobic
The right side obtains seed liquor.
3) it ferments
According to the inoculum concentration of 0.8% (v/v), above-mentioned seed liquor is inoculated in above-mentioned fermentation medium and carries out fermentation training
It supports, control fermentation temperature is 30 DEG C or so, and revolving speed is 120r/min or so, and fermentation period is 10 days or so, obtains loquat fermentation
Product.
Using People's Republic of China's agricultural industry criteria NY/T 2277-2012 method in above-mentioned loquat fermented product
Organic acid detected, the results are shown in Table 2.
Embodiment 3
One, culture medium is prepared
1) basal medium
By 12.0g casein peptone, 8.0g beef extract, 6.0g yeast powder, 4.0g glucose, 4.0g sodium acetate, 3.0g lemon
Sour diammonium, 0.5g Tween 80,1.0g K2HPO4、0.3g MgSO4·7H2O、0.01g MnSO4·H2O、12.0g CaCO3Dissolution
In 1L distilled water, pH value is adjusted to 6.5, basal medium (when preparing solid medium, 20g agar is added) is made.
2) fermentation medium
It squeezes the juice to dragon fruit, obtains dragon fruit Normal juice；Distilled water is added into dragon fruit Normal juice to size mixing, controls
Dragon fruit Normal juice and the volume ratio of distilled water are 1:3, and Dragonfruit Juice is made.
30g maltose, 5g ammonium chloride, 3g potassium dihydrogen phosphate and 12.0g CaCO are added into the above-mentioned Dragonfruit Juice of 1L3, adjust
PH value is saved to 6.5 or so, obtains fermentation medium.
Two, it ferments
1) actication of culture
By the lactobacillus plantarum being stored on inclined-plane (Lactobacillus plantarum) strain (CGMCC
No.14812 it) is inoculated in above-mentioned total amount of binder culture medium, stationary culture 18h or so, obtains activated spawn under anaerobic.
2) seed liquor is prepared
The strain of above-mentioned activation is inoculated in aforesaid liquid basal medium, stationary culture is left for 24 hours under anaerobic
The right side obtains seed liquor.
3) it ferments
According to the inoculum concentration of 0.9% (v/v), above-mentioned seed liquor is inoculated in above-mentioned fermentation medium and carries out fermentation training
It supports, control fermentation temperature is 32 DEG C or so, and revolving speed is 150r/min or so, and fermentation period is 15 days or so, obtains dragon fruit hair
Ferment product.
It adopts to ferment to above-mentioned dragon fruit using People's Republic of China's agricultural industry criteria NY/T 2277-2012 method and make
Organic acid in product is detected, and the results are shown in Table 3.
Embodiment 4
One, culture medium is prepared
1) basal medium
By 10.0g casein peptone, 10.0g beef extract, 5.0g yeast powder, 5.0g glucose, 5.0g sodium acetate, 2.0g lemon
Sour diammonium, 1.0g Tween 80,2.0g K2HPO4、0.2g MgSO4·7H2O、0.05g MnSO4·H2O、10.0g CaCO3Dissolution
In 1L distilled water, pH value is adjusted to 6.8, basal medium (when preparing solid medium, 20g agar is added) is made.
2) fermentation medium
It squeezes the juice to mulberry fruit, obtains raw muberry juice；Distilled water is added into raw muberry juice to size mixing, control mulberry fruit is former
The volume ratio of juice and distilled water is 1:1, and mulberry juice is made.
10g sucrose, 4g ammonium nitrate, 1g manganese sulfate, 1g dipotassium hydrogen phosphate and 10.0g are added into the above-mentioned mulberry juice of 1L
CaCO3, pH value is adjusted to 6.8 or so, obtains fermentation medium.
Two, it ferments
1) actication of culture
By the lactobacillus plantarum being stored on inclined-plane (Lactobacillus plantarum) strain (CGMCC
No.14812 it) is inoculated in above-mentioned total amount of binder culture medium, stationary culture controls for 24 hours under anaerobic, obtains activated spawn.
2) seed liquor is prepared
The strain of above-mentioned activation is inoculated in aforesaid liquid basal medium, stationary culture is left for 24 hours under anaerobic
The right side obtains seed liquor.
3) it ferments
According to the inoculum concentration of 1% (v/v), above-mentioned seed liquor is inoculated in above-mentioned fermentation medium and carries out fermented and cultured,
Controlling fermentation temperature is 35 DEG C or so, and revolving speed is 150r/min or so, and fermentation period is 7 days or so, obtains mulberry fruit fermented product.
Using People's Republic of China's agricultural industry criteria NY/T 2277-2012 method in above-mentioned mulberry fruit fermented product
Organic acid detected, the results are shown in Table 4.
Reference examples 1-4
Reference examples 1-4 using lactobacillus plantarum (Lactobacillus plantarum) reference culture ATCC 14917 as
Control, except using lactobacillus plantarum (Lactobacillus plantarum) reference culture ATCC14917 alternative embodiment 1-
Lactobacillus plantarum (Lactobacillus plantarum) bacterial strain CGMCC No.14812 and inoculum concentration in 4 is according to following
Illustrate except change, remaining is identical as embodiment 1-4 respectively:
In reference examples 1, according to the inoculum concentration of 3% (v/v), seed liquor is inoculated in fermentation medium and carries out fermentation training
It supports.
In reference examples 2, according to the inoculum concentration of 5% (v/v), seed liquor is inoculated in fermentation medium and carries out fermentation training
It supports.
In reference examples 3, according to the inoculum concentration of 1% (v/v), seed liquor is inoculated in fermentation medium and carries out fermentation training
It supports.
In reference examples 4, according to the inoculum concentration of 3% (v/v), seed liquor is inoculated in fermentation medium and carries out fermentation training
It supports.
The testing result of the fermented product of reference examples 1-4 is shown in Table 1 to table 4 respectively.
Organic acid testing result in the orange peel fermented product of 1 embodiment 1 of table and reference examples 1
Note: "/" expression is not detected.
Organic acid testing result in the loquat fermented product of 2 embodiment 2 of table and reference examples 2
Note: "/" expression is not detected.
Organic acid testing result in the dragon fruit fermented product of 3 embodiment 3 of table and reference examples 3
Note: "/" expression is not detected.
Organic acid testing result in the mulberry fruit fermented product of 4 embodiment 4 of table and reference examples 4
Note: "/" expression is not detected.
Table 1 to table 4 the result shows that:
Relative to unfermentable raw material juice and utilize lactobacillus plantarum (Lactobacillus plantarum) reference culture
The fermented product that ATCC 14917 ferments utilizes lactobacillus plantarum (Lactobacillus plantarum) of the invention
In the fermented product that (CGMCC No.14812) ferments, the content of organic acid is significantly improved.
Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Pipe present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that: its according to
So be possible to modify the technical solutions described in the foregoing embodiments, or to some or all of the technical features into
Row equivalent replacement；And these are modified or replaceed, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of lactobacillus plantarum (Lactobacillus plantarum), which is characterized in that deposit number CGMCC
No.14812。
2. application of the lactobacillus plantarum (Lactobacillus plantarum) described in claim 1 on production organic acid.
3. a kind of fermented product production method, which is characterized in that use lactobacillus plantarum described in claim 1
(Lactobacillus plantarum) ferments to raw material.
4. fermented product production method according to claim 3, which is characterized in that the raw material is selected from fruit and medicine food is same
At least one of source class raw material.
5. fermented product production method according to claim 4, which is characterized in that the fruit includes orange, loquat, fire
Imperial fruit and mulberry fruit, the integration of drinking and medicinal herbs class raw material include Poria cocos, Radix Glycyrrhizae, dark plum and chrysanthemum.
6. according to any fermented product production method of claim 3 to 5, which comprises the steps of:
It squeezed the juice, sized mixing to the raw material, obtain raw material juice；
0-80g carbon source, 3-8g nitrogen source, 1-3g inorganic salts and 5-12g calcium carbonate are added into raw material juice described in every liter, then adjusts
PH value obtains fermentation medium to 5.5-6.8；
The lactobacillus plantarum (Lactobacillus plantarum) is seeded to the fermentation by 0.5-1.5% (v/v) to train
It supports and ferments in base, obtain fermented product.
7. according to any fermented product production method of claim 3 to 5, which is characterized in that the temperature of the fermentation is
20-35 DEG C, revolving speed 60-150r/min, fermentation period is 7-30 days.
8. a kind of fermented product, which is characterized in that produced according to any fermented product production method of claim 3 to 7
It obtains.
9. fermented product according to claim 8, which is characterized in that contain organic acid in the fermented product.
10. fermented product according to claim 9, which is characterized in that the organic acid includes lactic acid, acetic acid, propionic acid, fourth
At least one of acid, malic acid, tartaric acid, oxalic acid and citric acid.
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