CN109628350B - Lactobacillus plantarum and application thereof - Google Patents
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Abstract
The invention provides lactobacillus plantarum and application thereof. The Lactobacillus plantarum (Lactobacillus plantarum) has a preservation number of CGMCC No. 14812. The lactobacillus plantarum provided by the invention can be used for improving organic acid in raw materials, and the lactobacillus plantarum can be used for fermenting the raw materials, wherein the raw materials can be selected from at least one of fruits. The lactobacillus plantarum provided by the invention is used for fermenting raw materials, so that the content of organic acid in a fermented product can be increased.
Description
Technical Field
The invention relates to lactic acid bacteria, and particularly relates to lactobacillus plantarum and application thereof.
Background
The organic acid is an acid containing carboxyl in the molecule, is widely present in animals, plants and microorganisms, and mainly comprises lactic acid, acetic acid, propionic acid, butyric acid, malic acid, tartaric acid, oxalic acid, citric acid and the like. Many organic acids can directly participate in biochemical reaction, and some of the organic acids have the effects of bacteriostasis, inflammation diminishing, virus resisting, mutation resisting, cancer resisting and the like; some of the medicine can increase coronary artery blood flow, inhibit the generation of lipid peroxide in brain tissue, soften blood vessel, promote the absorption of calcium and iron elements; some also have the effects of helping gastric juice digest fat and protein, preventing diseases and promoting metabolism, thereby being beneficial to the health of human bodies.
Lactic Acid Bacteria (LAB) are a general term for a group of bacteria that are capable of producing large amounts of lactic acid using fermentable carbohydrates. A large number of researches show that the lactobacillus can regulate normal flora of gastrointestinal tracts of organisms, keep micro-ecological balance, improve the digestibility and the biovalue of food, reduce serum cholesterol, control endotoxin, inhibit growth and propagation of putrefying bacteria in intestinal tracts and generation of putrefying products, produce nutrient substances and stimulate tissue development, thereby having effects on the nutritional state, the physiological function, cell infection, drug effect, toxic reaction, immune reaction, tumorigenesis, aging process, sudden emergency reaction and the like of the organisms.
Lactic acid bacteria produce substances such as organic acids by fermentation and thus exert their special physiological functions. The lactobacillus can decompose lactose into glucose and galactose, and the glucose is converted into small molecular compounds such as lactic acid by fermentation, which is helpful for the development of brain tissue and nervous system of children. Lactic acid bacteria have phosphoproteases and are capable of decomposing α -casein into fine cheese fatty peptides, amino acids, and the like, thereby improving the digestibility of proteins. The lactobacillus fermentation can degrade part of fat in a small amount, is easy to digest and can increase the content of free fatty acid and volatile fatty acid in milk. Lactic acid bacteria consume part of vitamins in the metabolic process, and simultaneously synthesize B vitamins such as folic acid and the like. The organic acid generated after lactobacillus fermentation can improve the utilization rate of elements such as calcium, phosphorus, iron and the like, promote the absorption of iron and VD, and has stronger bacteriostatic action.
At present, the content of organic acids in plants and microorganisms is relatively low, and therefore a method capable of increasing the content of organic acids is desired.
Disclosure of Invention
The invention provides lactobacillus plantarum and application thereof, and the lactobacillus plantarum is used for fermenting raw materials and can improve the content of organic acid in a fermented product.
The invention provides Lactobacillus plantarum (Lactobacillus plantarum) with a preservation number of CGMCC No. 14812.
The Lactobacillus plantarum (CGMCC No.14812) of the invention has the phenotypic characteristics of: g +, rod shape, round colony, milky white color, and regular edge.
The Lactobacillus plantarum (CGMCC No.14812) is obtained by separating and screening naturally fermented fruit and vegetable enzymes through a culturable method, and the taxonomic status is determined by the aid of the phylogenetic analysis of a 16S rRNA gene sequence and a pheS gene and by combination of phenotypic characteristics.
The growth conditions of Lactobacillus plantarum (CGMCC No.14812) of the present invention may be conventional conditions, and the growth conditions may be, for example: the growth temperature is 30-37 ℃, the facultative anaerobic culture medium is anaerobic, and the growth time is 24-48 h.
The Lactobacillus plantarum (CGMCC No.14812) of the present invention is capable of producing organic acids by fermentation, and thus can be used for related applications.
The invention also provides application of the Lactobacillus plantarum (CGMCC No.14812) in producing organic acid.
The invention does not strictly limit the application mode of the Lactobacillus plantarum (CGMCC No.14812) in producing the organic acid, can produce the fermentation product containing the organic acid, and can extract the organic acid from the fermentation product according to the requirement to provide downstream application.
The invention also provides a production method of a fermented product, which adopts the Lactobacillus plantarum (CGMCC No.14812) to ferment raw materials.
The raw materials are not strictly limited, and the method is only favorable for producing the organic acid by the Lactobacillus plantarum (CGMCC No. 14812).
Specifically, the raw materials can be selected from at least one of fruits and medicinal and edible raw materials. Furthermore, the fruits comprise oranges, loquats, dragon fruits, mulberries and the like, and the medicinal and edible raw materials comprise poria cocos, liquorice, dark plums, chrysanthemum and the like.
When the Lactobacillus plantarum (CGMCC No.14812) is used to ferment a raw material, the fermentation medium and the fermentation conditions are not strictly limited as long as the Lactobacillus plantarum (CGMCC No.14812) is favorable for the production of an organic acid.
In a specific embodiment of the present invention, the method for producing a fermentation product may comprise the steps of:
juicing and mixing the raw materials to obtain raw juice;
adding 0-80g of carbon source, 3-8g of nitrogen source, 1-3g of inorganic salt and 5-12g of calcium carbonate into each liter of the raw material juice, and then adjusting the pH value to 5.5-6.8 to obtain a fermentation medium;
inoculating the Lactobacillus plantarum (Lactobacillus plantarum) into the fermentation medium according to the ratio of 0.5-1.5% (v/v) for fermentation to obtain a fermentation product.
In the invention, the raw materials can be squeezed in a conventional manner to obtain raw juice; in addition, the raw juice can be mixed according to actual needs. Specifically, size mixing can be performed by using distilled water, wherein the volume ratio of raw material juice to distilled water can be controlled to be 1: (1-5), thereby preparing a raw material juice.
The carbon source, nitrogen source and inorganic salt are not particularly limited in the present invention, as long as they are suitable for the production of the organic acid by the above-mentioned Lactobacillus plantarum. Specifically, at least one of sucrose, glucose, maltose, etc. may be used as the carbon source, at least one of peptone, beef extract, ammonium chloride, ammonium nitrate, potassium nitrate, etc. may be used as the nitrogen source, and at least one of manganese sulfate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, etc. may be used as the inorganic salt.
Further, calcium carbonate may be added to the raw juice. With the progress of fermentation, the strains continuously produce acid, the pH value can be reduced, and the growth and fermentation of the strains are possibly influenced; therefore, the consumption of H can be realized by adding calcium carbonate according to the needs+And stabilizing pH.
The process of producing organic acid by using the Lactobacillus brevis (Lactobacillus plantarum) also comprises the operations of strain activation and seed liquid preparation, and the operation of inoculating the seed liquid to a fermentation culture medium for fermentation. The fermentation conditions in the present invention are not particularly limited as long as they are suitable for the production of organic acids by the above-mentioned Lactobacillus plantarum. Specifically, the fermentation temperature can be 20-35 ℃, the rotation speed can be 60-150r/min, and the fermentation period can be 7-30 days. The research of the inventor shows that compared with the similar strains, such as the Lactobacillus plantarum standard strain ATCC14917, the Lactobacillus plantarum (CGMCC No.14812) provided by the invention has the advantages of improved organic acid content in the fermentation product, reduced required inoculation amount and obvious production efficiency and cost.
The invention also provides a fermented product produced by the production method of any one of the fermented products.
The fermentation product of the present invention contains an organic acid. Further, the organic acid includes at least one of lactic acid, acetic acid, propionic acid, butyric acid, malic acid, tartaric acid, oxalic acid, citric acid, and the like.
The Lactobacillus plantarum (Lactobacillus plantarum) is preserved in 16 months 10 and 2017 in China general microbiological culture Collection center (CGMCC for short), the address of the Lactobacillus plantarum is No. 3 of Xilu No.1 of Beijing area and the rising area, and the preservation number is CGMCC No. 14812.
The Lactobacillus plantarum (CGMCC No.14812) provided by the invention can ferment to produce organic acid, and when the Lactobacillus plantarum is used for fermenting raw materials, the content of the organic acid in a fermented product can be increased.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
Firstly, preparing a culture medium
1) Basic culture medium
Mixing casein peptone 10.0g, beef extract 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, Tween-80 1.0g, and Tween-2.0 g 2.0g K2HPO4、0.2g MgSO4·7H2O、0.05g MnSO4·H2O、10.0g CaCO3Dissolving in 1L distilled water, adjusting pH to 6.8, and making into basal culture medium (20 g agar is added when preparing solid culture medium).
2) Fermentation medium
Juicing orange peel to obtain orange peel raw juice; adding distilled water into the orange peel normal juice for size mixing, and controlling the volume ratio of the orange peel normal juice to the distilled water to be 1: 1, preparing the orange peel juice.
To 1L of the above orange peel juice were added 50g of glucose, 6g of peptone, 1.2g of manganese sulfate and 10.0g of CaCO3And adjusting the pH value to about 6.8 to obtain a fermentation culture medium.
Secondly, fermentation
1) Strain activation
Inoculating Lactobacillus plantarum (CGMCC No.14812) preserved on the inclined plane into the solid basal culture medium, and performing static culture under anaerobic condition for about 24h to obtain activated strain.
2) Preparation of seed liquid
Inoculating the activated strain into the liquid basal culture medium, and performing static culture for about 24h under anaerobic conditions to obtain seed liquid.
3) Fermentation of
Inoculating the seed liquid into the fermentation culture medium according to the inoculation amount of 1% (v/v) for fermentation culture at the fermentation temperature of about 35 ℃, the rotation speed of about 150r/min and the fermentation period of about 7 days to obtain the orange peel fermented product.
The organic acid in the orange peel fermentation product is detected by adopting a method of agricultural industry standard NY/T2277-.
Example 2
Firstly, preparing a culture medium
1) Basic culture medium
Mixing 8.0g of cheesePeptone, 12.0g beef extract, 4.0g yeast powder, 6.0g glucose, 6.0g sodium acetate, 1.0g diammonium citrate, 1.5g tween 80, 3.0g K2HPO4、0.1g MgSO4·7H2O、0.1g MnSO4·H2O、8.0g CaCO3Dissolving in 1L distilled water, adjusting pH to 6.0, and making into basal culture medium (20 g agar is added when preparing solid culture medium).
2) Fermentation medium
Squeezing fructus Eriobotryae to obtain fructus Eriobotryae natural juice; adding distilled water into the loquat raw juice for size mixing, and controlling the volume ratio of the loquat raw juice to the distilled water to be 1: 2, preparing the loquat juice.
Adding 60g glucose, 3g peptone, 2.0g manganese sulfate and 8.0g CaCO into 1L of the above loquat juice3And adjusting the pH value to about 6.0 to obtain a fermentation culture medium.
Secondly, fermentation
1) Strain activation
Inoculating Lactobacillus plantarum (CGMCC No.14812) strain preserved on the inclined plane into the solid basal culture medium, and performing static culture under anaerobic condition for about 18h to obtain activated strain.
2) Preparation of seed liquid
Inoculating the activated strain into the liquid basal culture medium, and performing static culture for about 24h under anaerobic conditions to obtain seed liquid.
3) Fermentation of
Inoculating the seed solution into the fermentation culture medium according to the inoculation amount of 0.8% (v/v), and performing fermentation culture at 30 deg.C or so and at 120r/min or so for 10 days or so to obtain fermented loquat product.
The organic acid in the loquat fermentation product is detected by adopting a method of agricultural industry standard NY/T2277-.
Example 3
Firstly, preparing a culture medium
1) Basic culture medium
Mixing 12.0g casein peptone, 8.0g beef extract,6.0g of yeast powder, 4.0g of glucose, 4.0g of sodium acetate, 3.0g of diammonium citrate, 0.5g of Tween 80 and 1.0g K2HPO4、0.3g MgSO4·7H2O、0.01g MnSO4·H2O、12.0g CaCO3Dissolving in 1L distilled water, adjusting pH to 6.5, and making into basal culture medium (20 g agar is added when preparing solid culture medium).
2) Fermentation medium
Squeezing the dragon fruit to obtain raw dragon fruit juice; adding distilled water into the raw dragon fruit juice for size mixing, and controlling the volume ratio of the raw dragon fruit juice to the distilled water to be 1: and 3, preparing the dragon fruit juice.
Adding maltose 30g, ammonium chloride 5g, potassium dihydrogen phosphate 3g and CaCO 12.0g into dragon fruit juice 1L3And adjusting the pH value to about 6.5 to obtain a fermentation culture medium.
Secondly, fermentation
1) Strain activation
Inoculating Lactobacillus plantarum (CGMCC No.14812) strain preserved on the inclined plane into the solid basal culture medium, and performing static culture under anaerobic condition for about 18h to obtain activated strain.
2) Preparation of seed liquid
Inoculating the activated strain into the liquid basal culture medium, and performing static culture for about 24h under anaerobic conditions to obtain seed liquid.
3) Fermentation of
Inoculating the seed liquid into the fermentation culture medium according to the inoculation amount of 0.9% (v/v) for fermentation culture at the fermentation temperature of about 32 ℃, the rotation speed of about 150r/min and the fermentation period of about 15 days to obtain the dragon fruit fermented product.
The organic acid in the dragon fruit fermentation product is detected by adopting a method of agricultural industry standard NY/T2277-.
Example 4
Firstly, preparing a culture medium
1) Basic culture medium
Mixing casein peptone 10.0g and beef 10.0gCream, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, diammonium citrate 2.0g, Tween 80 1.0g, and Tween 2.0g K2HPO4、0.2g MgSO4·7H2O、0.05g MnSO4·H2O、10.0g CaCO3Dissolving in 1L distilled water, adjusting pH to 6.8, and making into basal culture medium (20 g agar is added when preparing solid culture medium).
2) Fermentation medium
Squeezing Mori fructus to obtain Mori fructus natural juice; adding distilled water into the mulberry raw juice for size mixing, and controlling the volume ratio of the mulberry raw juice to the distilled water to be 1: 1, preparing the mulberry juice.
Adding 10g sucrose, 4g ammonium nitrate, 1g manganese sulfate, 1g dipotassium hydrogen phosphate and 10.0g CaCO into 1L of the above Mori fructus juice3And adjusting the pH value to about 6.8 to obtain a fermentation culture medium.
Secondly, fermentation
1) Strain activation
Inoculating Lactobacillus plantarum (CGMCC No.14812) preserved on the inclined plane into the solid basal culture medium, and performing static culture under anaerobic condition for about 24h to obtain activated strain.
2) Preparation of seed liquid
Inoculating the activated strain into the liquid basal culture medium, and performing static culture for about 24h under anaerobic conditions to obtain seed liquid.
3) Fermentation of
Inoculating the seed solution into the fermentation culture medium according to the inoculation amount of 1% (v/v), and performing fermentation culture at about 35 deg.C and rotation speed of about 150r/min for about 7 days to obtain Mori fructus fermented product.
The organic acid in the mulberry fermented product is detected by adopting a method of agricultural industry standard NY/T2277-.
Comparative examples 1 to 4
Comparative examples 1 to 4 were conducted in the same manner as in examples 1 to 4 except that the standard strain of Lactobacillus plantarum ATCC14917 was used as a control, the standard strain of Lactobacillus plantarum ATCC14917 was used in place of the Lactobacillus plantarum CGMCC No.14812 in examples 1 to 4, and the inoculation amount was changed as described below:
in comparative example 1, the seed solution was inoculated into a fermentation medium in an amount of 3% (v/v) for fermentation culture.
In comparative example 2, the seed solution was inoculated into a fermentation medium in an amount of 5% (v/v) for fermentation culture.
In comparative example 3, the seed solution was inoculated into the fermentation medium in an amount of 1% (v/v) for fermentation culture.
In comparative example 4, the seed solution was inoculated into the fermentation medium in an amount of 3% (v/v) for fermentation culture.
The results of measurement of the fermented products of comparative examples 1 to 4 are shown in tables 1 to 4, respectively.
Table 1 results of organic acid detection in fermented products of orange peels of example 1 and comparative example 1
Note: "/" indicates no detection.
Table 2 results of organic acid detection in loquat fermented products of example 2 and comparative example 2
Note: "/" indicates no detection.
Table 3 detection results of organic acids in the dragon fruit fermented products of example 3 and comparative example 3
Note: "/" indicates no detection.
Table 4 results of organic acid assay in fermented morous alba products of example 4 and comparative example 4
Note: "/" indicates no detection.
The results in tables 1 to 4 show that:
compared with the unfermented raw material juice and the fermented product fermented by the Lactobacillus plantarum (CGMCC No.14812), the content of organic acid in the fermented product fermented by the Lactobacillus plantarum (CGMCC No.14812) is obviously improved.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (8)
1. A Lactobacillus plantarum (A)Lactobacillus plantarum) The preservation number is CGMCC number 14812.
2. Lactobacillus plantarum (b) according to claim 1Lactobacillus plantarum) Application in increasing total organic acid content in fruit juice.
3. A method for producing a fermented product, which comprises using the Lactobacillus plantarum (L) according to claim 1Lactobacillus plantarum) Fermenting the raw materials; the fermentation product contains organic acid; the raw material is selected from fruits.
4. The method for producing fermented products according to claim 3, wherein the fruits include oranges, loquats, pitaya and morous alba.
5. The process for the production of a fermentation product according to any one of claims 3 to 4, comprising the steps of:
juicing and mixing the raw materials to obtain raw juice;
adding 0-80g of carbon source, 3-8g of nitrogen source, 1-3g of inorganic salt and 5-12g of calcium carbonate into each liter of the raw material juice, and then adjusting the pH value to 5.5-6.8 to obtain a fermentation medium;
the lactobacillus plantarum (a)Lactobacillus plantarum) Inoculating the strain into the fermentation medium according to the ratio of 0.5-1.5% (v/v) for fermentation to obtain a fermentation product.
6. The method for producing a fermented product according to any one of claims 3 to 4, wherein the fermentation temperature is 20 to 35 ℃, the rotation speed is 60 to 150r/min, and the fermentation period is 7 to 30 days.
7. A fermented product produced by the method for producing a fermented product according to any one of claims 3 to 6; the fermentation product contains organic acid.
8. The fermentation product of claim 7, wherein the organic acid comprises at least one of lactic acid, butyric acid, malic acid, tartaric acid, oxalic acid, and citric acid.
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| KR101380920B1 (en) * | 2013-01-15 | 2014-04-02 | 계명대학교 산학협력단 | Fermented jam comprising fermented opuntia humifusa fruit by lactic acid bacteria |
| CN104357353A (en) * | 2014-11-03 | 2015-02-18 | 郑州大学 | Application of lactobacillus plantarum in making fermented fruit and vegetable juice |
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| KR101380920B1 (en) * | 2013-01-15 | 2014-04-02 | 계명대학교 산학협력단 | Fermented jam comprising fermented opuntia humifusa fruit by lactic acid bacteria |
| CN104357353A (en) * | 2014-11-03 | 2015-02-18 | 郑州大学 | Application of lactobacillus plantarum in making fermented fruit and vegetable juice |
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| 1株植物乳杆菌发酵豆粕产有机酸的研究;毛银 等;《食品与发酵工业》;20170720;第43-48页 * |
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