CN102260720A - Method for producing gamma-aminobutyric acid by using Lactobacillus fructivorans - Google Patents
Method for producing gamma-aminobutyric acid by using Lactobacillus fructivorans Download PDFInfo
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- CN102260720A CN102260720A CN2011102029227A CN201110202922A CN102260720A CN 102260720 A CN102260720 A CN 102260720A CN 2011102029227 A CN2011102029227 A CN 2011102029227A CN 201110202922 A CN201110202922 A CN 201110202922A CN 102260720 A CN102260720 A CN 102260720A
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- lactobacillus fructivorans
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Abstract
The invention discloses a method for producing gamma-aminobutyric acid by using Lactobacillus fructivorans. The method comprises the following steps of: (1) activating, namely dissolving bacterial blocks of the Lactobacillus fructivorans, and inoculating on an Man-Rogosa-Sharpe (MRS) liquid culture medium and culturing and activating; and (2) fermenting, namely inoculating the activated Lactobacillus fructivorans in the MRS culture medium containing 1 percent of sodium glutamate, and fermenting and culturing for 2 to 10 days under the condition that the temperature is 30 to 35 DEG and the pH value is 3.0 to 4.0 to obtain culture solution containing the gamma-aminobutyric acid. Fermentation raw materials are easily obtained, few byproducts are produced in the transformation process, harmful substances are avoided, and the product has the advantages of low cost, high content of the gamma-aminobutyric acid and the like, and can be safely used for foods, applied to foods, medicines and feeds, and directly eaten or used as a functional base material for enriching the gamma-aminobutyric acid according to production requirements. In the prior art, any report on the production of the gamma-aminobutyric acid by using the Lactobacillus fructivorans has not been found yet.
Description
Technical field
The present invention relates to the γ-An Jidingsuan technical field, particularly relate to a kind of method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan.
Background technology
(γ-aminobutyric acid GABA) claims aminobutyric acid again to γ-An Jidingsuan, is the natural amino acid that a kind of nonprotein is formed.GABA has many important physical functions, as bring high blood pressure down, calmness, hypermnesis, adjusting respiratory system, anxiety, especially to involutional insomnia, constrain and self imbalance curative effect good.In addition, GABA also has the human liver of improvement function, the activation renal function, and prevention of obesity prevents skin aging, promotes alcohol metabolism, effects such as smelly effect disappear.Along with the physiological function of GABA is constantly studied and is illustrated, developed into a kind of novel functional component, very big potentiality are arranged in the exploitation of functional foodstuff, be applied to industries such as medicine, food just gradually.At present, the feasibility of microorganisms producing GABA has obtained confirmation, and various lactobacillus such as short lactobacillus, Lactococcus lactis, plant lactobacillus are produced GABA all report.
Summary of the invention
Technical problem to be solved by this invention provides a kind of method that adopts Lactobacillus fructivorans to produce γ-An Jidingsuan, and cost is low, the content height of γ-An Jidingsuan in the gained nutrient solution.
In order to solve the problems of the technologies described above, the present invention adopts following technical scheme:
The present invention utilizes Lactobacillus fructivorans to produce the method for γ-An Jidingsuan, comprises the steps:
(1) activation: cultivate activation on the MRS liquid nutrient medium with being inoculated in after the dissolving of Lactobacillus fructivorans bacterium piece;
(2) fermentation: the Lactobacillus fructivorans after the activation is inoculated in and contains the 1%(volume) in the MRS substratum of Sodium Glutamate, cultivated 2～10 days at 30～35 ℃, pH 3.0～4.0 condition bottom fermentations, promptly contain γ-An Jidingsuan in the gained nutrient solution.
In order to obtain a large amount of Lactobacillus fructivorans, can will ferment again after the Lactobacillus fructivorans enlarged culturing after the activation, the enlarged culturing concrete operations are as follows: will activate good Lactobacillus fructivorans and be inoculated in the liquid MRS substratum 30 ℃ of enlarged culturing.
Preferably, the aforementioned method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan is:
(1) activation: with sterilized water or nutrient solution dissolving Lactobacillus fructivorans bacterium piece, be inoculated on the MRS liquid nutrient medium, cultivate activation for 30 ℃;
(2) inoculum size enlarged culturing: will activate good Lactobacillus fructivorans with the 2%(volume) is inoculated in the MRS substratum, cultivates 48 hours for 30 ℃;
(3) fermentation: the Lactobacillus fructivorans after the activation is inoculated in 10% inoculum size and contains the 1%(volume) in the MRS substratum of Sodium Glutamate, cultivated 2～10 days at 30～35 ℃, pH 3.0～4.0 condition bottom fermentations, promptly contain γ-An Jidingsuan in the gained nutrient solution.
Used Lactobacillus fructivorans is Lactobacillus fructivorans Lactobacillus fructivorans in the preceding method, and deposit number is CGMCC No.1.2427.
MRS substratum used in the preceding method is: peptone 10g, extractum carnis 10g, yeast extract paste 5g, glucose 5g, sodium acetate 5g, Trisodium Citrate 0.05g, dipotassium hydrogen phosphate 2g, sal epsom 0.2g, manganous sulfate 0.05g, tween 80 1ml, distilled water 1000ml, mix the back and adjust pH5.0,121 ℃ of high pressure, sterilization 20min.
Adopt the aforementioned method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan, the content of γ-An Jidingsuan is 2.15g/L～6.35g/L in the final fermentation culture that obtains.
Compared with prior art, the target that the present invention adopts Lactobacillus fructivorans to produce γ-An Jidingsuan is first cultivated, with Sodium Glutamate is main substrate, Lactobacillus fructivorans (Lactobacillus fructivorans) fermentation that utilization has food safety makes the product that contains γ-An Jidingsuan, fermentation raw material is easy to obtain, by product is few in the conversion process, unharmful substance exists, has the low and alpha-aminobutyric acid content height of cost, can be used safely in advantages such as food, Lactobacillus fructivorans is as food-grade microorganisms simultaneously, it is inherent advantage milk-acid bacteria in the fruits and vegetables fermentation, leavening property, flavor characteristics and security are good, the γ-An Jidingsuan tunning that contains with its production can be applicable to food, medicine, in the feed, can directly eat or according to producing the function base-material of needs as enriching gamma-aminobutyric.Yet there are no the report of producing γ-An Jidingsuan with Lactobacillus fructivorans in the prior art.
Embodiment
1, bacterial classification: Lactobacillus fructivorans (Lactobacillus fructivorans) bacterial classification, derive from China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC), deposit number is: CGMCC No.1.2427, the freeze-drying Lactobacillus fructivorans that the ampoul tube sealing is preserved is in refrigerator 4-6 ℃ preservation.
2, actication of culture: need activation before bacterial classification uses.The activation concrete operation method: earlier with 70% alcohol cotton wiping ampoule top, heat is burnt at the ampoul tube top, be stained with cold water with aseptic cotton carrier, wipe a circle at the top, crackle appears in the top, with frustrating cutter or the tapping of tweezers neck once, strikes down rimose ampoul tube.With 1ml sterilized water or nutrient solution dissolving 10mg bacterium piece, use aseptic straw to move on the fresh MRS liquid nutrient medium, 30 ℃ are carried out thermophilic and cultivate.
3, enlarged culturing: Lactobacillus fructivorans is enlarged culturing in the MRS substratum, and concrete operations are: will activate good Lactobacillus fructivorans, with the 2%(volume) inoculum size be inoculated in the liquid MRS substratum, cultivate 48h for 30 ℃.
4, fermentation culture: with the bacterial classification of enlarged culturing with the 10%(volume) inoculum size be seeded to contain the 1%(volume) in the MRS substratum of Sodium Glutamate, at 30-35 ℃, pH 3.0-4.0(by dripping HCl or NaOH control) cultivation and fermentation 2 days under the condition, alpha-aminobutyric acid content reaches 2.15g/L in the gained nutrient solution, continue fermentation after 10 days, alpha-aminobutyric acid content reaches 6.35g/L in the gained nutrient solution.
Used MRS substratum is in the aforesaid method: peptone 10g, extractum carnis 10g, yeast extract paste 5g, glucose 5g, sodium acetate 5g, Trisodium Citrate 0.05g, dipotassium hydrogen phosphate 2g, sal epsom 0.2g, manganous sulfate 0.05g, tween 80 1ml, distilled water 1000ml, mix the back and adjust pH5.0,121 ℃ of high pressure, sterilization 20min.
Claims (6)
1. a method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan is characterized in that: comprise the steps:
(1) activation: cultivate activation on the MRS liquid nutrient medium with being inoculated in after the dissolving of Lactobacillus fructivorans bacterium piece;
(2) fermentation: the Lactobacillus fructivorans after the activation is inoculated in and contains the 1%(volume) in the MRS substratum of Sodium Glutamate, cultivated 2～10 days at 30～35 ℃, pH 3.0～4.0 condition bottom fermentations, promptly contain γ-An Jidingsuan in the gained nutrient solution.
2. according to the described method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan of claim 1, it is characterized in that: ferment again after the Lactobacillus fructivorans enlarged culturing after the activation, the enlarged culturing concrete operations are as follows: will activate good Lactobacillus fructivorans and be inoculated in the liquid MRS substratum 30 ℃ of enlarged culturing.
3. according to the described method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan of claim 2, it is characterized in that: comprise the steps:
(1) activation: with sterilized water or nutrient solution dissolving Lactobacillus fructivorans bacterium piece, be inoculated on the MRS liquid nutrient medium, cultivate activation for 30 ℃;
(2) inoculum size enlarged culturing: will activate good Lactobacillus fructivorans with the 2%(volume) is inoculated in the MRS substratum, cultivates 48 hours for 30 ℃;
(3) fermentation: the Lactobacillus fructivorans after the activation is inoculated in 10% inoculum size and contains the 1%(volume) in the MRS substratum of Sodium Glutamate, cultivated 2～10 days at 30～35 ℃, pH 3.0～4.0 condition bottom fermentations, promptly contain γ-An Jidingsuan in the gained nutrient solution.
4. according to the arbitrary described method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan of claim 1 to 3, it is characterized in that: the deposit number of described Lactobacillus fructivorans Lactobacillus fructivorans is CGMCC No.1.2427.
5. according to the arbitrary described method of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan of claim 1 to 3, it is characterized in that: described MRS substratum is: peptone 10g, extractum carnis 10g, yeast extract paste 5g, glucose 5g, sodium acetate 5g, Trisodium Citrate 0.05g, dipotassium hydrogen phosphate 2g, sal epsom 0.2g, manganous sulfate 0.05g, tween 80 1ml, distilled water 1000ml, mix the back and adjust pH5.0,121 ℃ of high pressure, sterilization 20min.
6. according to the right 3 described methods of utilizing Lactobacillus fructivorans to produce γ-An Jidingsuan, it is characterized in that: the content of γ-An Jidingsuan is 2.15g/L～6.35g/L in the nutrient solution after the fermentation culture.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108208590A (en) * | 2017-12-29 | 2018-06-29 | 涂慧燕 | A kind of germinated rice rice flour of high gamma-aminobutyric acid and phytic acid and preparation method thereof |
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| CN1392262A (en) * | 2002-06-08 | 2003-01-22 | 江南大学 | Process for preparing food function factor gamma-amino-butyric acid |
| CN1683544A (en) * | 2005-03-07 | 2005-10-19 | 浙江大学 | Process for biologically synthesizing gamma-amino butyric acid |
| CN101333548A (en) * | 2008-06-26 | 2008-12-31 | 南昌大学 | Method for preparing gamma-aminobutyric acid by lactobacillus brevis |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1392262A (en) * | 2002-06-08 | 2003-01-22 | 江南大学 | Process for preparing food function factor gamma-amino-butyric acid |
| CN1683544A (en) * | 2005-03-07 | 2005-10-19 | 浙江大学 | Process for biologically synthesizing gamma-amino butyric acid |
| CN101333548A (en) * | 2008-06-26 | 2008-12-31 | 南昌大学 | Method for preparing gamma-aminobutyric acid by lactobacillus brevis |
Non-Patent Citations (2)
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| 《中国酿造》 20110430 范杰等 高产gamma-氨基丁酸乳酸菌的筛选及鉴定 39-41 1-6 , 第4期 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108208590A (en) * | 2017-12-29 | 2018-06-29 | 涂慧燕 | A kind of germinated rice rice flour of high gamma-aminobutyric acid and phytic acid and preparation method thereof |
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Application publication date: 20111130 |