Lactobacillus paracasei and uses thereof

Abstract

A Lactobacillus paracasei and uses thereof. The Lactobacillus paracasei has a deposit number of CGMCC No. 14813. The Lactobacillus paracasei can be used for increasing the amount of an organic acid in a raw material. The Lactobacillus paracasei can be used for fermenting the raw material, where the raw material can be selected from at least one of fruits, which can increase the content of the organic acid in a fermented product.

Claims

1. A method for producing an organic acid, the method comprising: inoculating a raw material with a Lactobacillus paracasei and fermenting said raw material and said Lactobacillus paracasei to produce said organic acid, wherein said fermenting is at a temperature of 20-35? C., a rotation speed of 60-150 r/min, and a fermentation period of 7-30 days; wherein said Lactobacillus paracasei has a deposit number of CGMCC No. 14813, wherein said organic acid produced by said Lactobacillus paracasei with the deposit number of CGMCC No. 14813 has an increased total content of an organic acid, as compared to an organic acid produced by an Lactobacillus paracasei strain ATCC 25302, and wherein said organic acid comprises at least three of lactic acid, acetic acid, propionic acid, butyric acid, malic acid, tartaric acid, oxalic acid, and citric acid.

2. The method for producing the organic acid of claim 1, wherein the raw material is selected from at least one of a fruit and a medicinal and edible homologous raw material.

3. The method for producing the organic acid of claim 2, wherein the fruit comprises grapes, blueberries, raspberries, and pears, and the medicinal and edible homologous raw material comprises mulberries, lilies, dark plums, and lotus seeds.

4. The method for producing the organic acid of claim 1, further comprising: juicing and mixing the raw material to obtain a raw material juice; adding 0-80 g of a carbon source, 3-8 g of a nitrogen source, 1-3 g of an inorganic salt and 8-12 g of calcium carbonate to the raw material juice per liter, and then adjusting pH value to 5.5-6.8 to obtain a fermented culture medium; inoculating the Lactobacillus paracasei with the deposit number of CGMCC No. 14813 into the fermented culture medium at a concentration of 0.5-1.5% (v/v) for fermenting to produce said organic acid.

Description

DESCRIPTION OF EMBODIMENTS

(1) In order to make the objectives, technical solutions, and advantages of the present disclosure clearer, the technical solutions in embodiments of the present disclosure will be described clearly and comprehensively in combination with the embodiments of the present disclosure. Apparently, the described embodiments are a part rather than all embodiments of the present disclosure. Based on the embodiments of the present disclosure, all other embodiments obtained by those ordinary skilled in the art without creative effort fall within the protection scope of the present disclosure.

Example 1

(2) I. Preparation of a Culture Medium

(3) 1) Basic Culture Medium

(4) 10.0 g of casein peptone, 10.0 g of beef extract, 5.0 g of yeast powder, 5.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diammonium citrate, 1.0 g of tween 80, 2.0 g of K.sub.2HPO.sub.4, 0.2 g of MgSO.sub.4.Math.7H.sub.2O, 0.05 g of MnSO.sub.4.Math.H.sub.2O, and 10.0 g of CaCO.sub.3 are dissolved in 1 L of distilled water, and the pH value is adjusted to 6.8, to obtain a basic culture medium (20 g of agar is added when a solid culture medium is prepared).

(5) 2) Fermented Culture Medium

(6) grapes are juiced to obtain a grape raw juice; distilled water is added to the grape raw juice for size mixing, and a volume ratio of the grape raw juice to distilled water is controlled to be 1:1 to obtain a grape juice.

(7) 40 g of glucose, 6 g of peptone, 2.5 g of manganese sulfate, and 10.0 g of CaCO.sub.3 are added to 1 L of the above grape juice, and the pH value is adjusted to about 6.8 to obtain a fermented culture medium.

(8) II. Fermentation

(9) 1) Activation of Strains

(10) Lactobacillus paracasei strains (CGMCC No. 14813) stored on an inclined plane are inoculated into the above solid basic culture medium, and then subjected to standing and culturing for about 24 hours under an anaerobic condition to obtain activated strains.

(11) 2) Preparation of a Seed Solution

(12) the above activated strains are inoculated into the above liquid basic culture medium, and then subjected to standing and culturing for about 24 h under the anaerobic condition to obtain a seed solution.

(13) 3) Fermentation

(14) according to the inoculation amount of 0.8% (v/v), the above seed solution is inoculated into the above fermented culture medium for fermenting and culturing, and the fermentation temperature is controlled to be at about 35? C., the rotation speed is controlled to be about 150 r/min, and the fermentation period is controlled to be about 7 days, to obtain a grape fermented product.

(15) The organic acid in the grape fermented product is detected by adopting the agricultural industry standard NY/T 2277-2012 method of the People's Republic of China, and the results are shown in Table 1.

Example 2

(16) I. Preparation of a Culture Medium

(17) 1) Basic Culture Medium

(18) 8.0 g of casein peptone, 12.0 g of beef extract, 4.0 g of yeast powder, 6.0 g of glucose, 6.0 g of sodium acetate, 1.0 g of diammonium citrate, 1.5 g of tween 80, 3.0 g of K.sub.2HPO.sub.4, 0.1 g of MgSO.sub.4.Math.7H.sub.2O, 0.1 g of MnSO.sub.4.Math.H.sub.2O, and 8.0 g of CaCO.sub.3 are dissolved in 1 L of distilled water, and the pH value is adjusted to 6.0, to obtain a basic culture medium (20 g of agar is added when a solid culture medium is prepared).

(19) 2) Fermented Culture Medium

(20) blueberries are juiced to obtain a blueberry raw juice; distilled water is added to the blueberry raw juice for size mixing, and a volume ratio of the blueberry raw juice to distilled water is controlled to be 1:2 to obtain a blueberry juice.

(21) 60 g of glucose, 4 g of peptone, 2.0 g of manganese sulfate, and 8.0 g of CaCO.sub.3 are added to 1 L of the above blueberry juice, and the pH value is adjusted to about 6.0 to obtain a fermented culture medium.

(22) II. Fermentation

(23) 1) Activation of Strains

(24) Lactobacillus paracasei strains (CGMCC No. 14813) stored on an inclined plane are inoculated into the above solid basic culture medium, and then subjected to standing and culturing for about 18 hours under an anaerobic condition to obtain activated strains.

(25) 2) Preparation of a Seed Solution

(26) the above activated strains are inoculated into the above liquid basic culture medium, and then subjected to standing and culturing for about 24 h under the anaerobic condition to obtain a seed solution.

(27) 3) Fermentation

(28) according to the inoculation amount of 0.8% (v/v), the above seed solution is inoculated into the above fermented culture medium for fermenting and culturing, and the fermentation temperature is controlled to be at about 30? C., the rotation speed is controlled to be about 120 r/min, and the fermentation period is controlled to be about 10 days, to obtain a blueberry fermented product.

(29) The organic acid in the blueberry fermented product is detected by adopting the agricultural industry standard NY/T 2277-2012 method of the People's Republic of China, and the results are shown in Table 2.

Example 3

(30) I. Preparation of a Culture Medium

(31) 1) Basic Culture Medium

(32) 12.0 g of casein peptone, 8.0 g of beef extract, 6.0 g of yeast powder, 4.0 g of glucose, 4.0 g of sodium acetate, 3.0 g of diammonium citrate, 0.5 g of tween 80, 1.0 g of K.sub.2HPO.sub.4, 0.3 g of MgSO.sub.4.Math.7H.sub.2O, 0.01 g of MnSO.sub.4.Math.H.sub.2O, and 12.0 g of CaCO.sub.3 are dissolved in 1 L of distilled water, and the pH value is adjusted to 6.5, to obtain a basic culture medium (20 g of agar is added when a solid culture medium is prepared).

(33) 2) Fermented Culture Medium

(34) raspberries are juiced to obtain a raspberry raw juice; distilled water is added to the raspberry raw juice for size mixing, and a the volume ratio of the raspberry raw juice to distilled water is controlled to be 1:3 to obtain a raspberry juice.

(35) 20 g of maltose, 5 g of ammonium chloride, 2 g of potassium dihydrogen phosphate, and 12.0 g of CaCO.sub.3 to 1 L of the above raspberry juice, and the pH value is adjusted to about 6.5 to obtain a fermented culture medium.

(36) II. Fermentation

(37) 1) Activation of Strains

(38) Lactobacillus paracasei strains (CGMCC No. 14813) stored on an inclined plane are inoculated into the above solid basic culture medium, and then subjected to standing and culturing for about 18 hours under an anaerobic condition to obtain activated strains.

(39) 2) Preparation of a Seed Solution

(40) the above activated strains are inoculated into the above liquid basic culture medium, and then subjected to standing and culturing for about 24 h under the anaerobic condition to obtain a seed solution.

(41) 3) Fermentation

(42) according to the inoculation amount of 1% (v/v), the above seed solution is inoculated into the above fermented culture medium for fermenting and culturing, and the fermentation temperature is controlled to be at about 32? C., the rotation speed is controlled to be about 150 r/min, and the fermentation period is controlled to be about 15 days, to obtain a raspberry fermented product.

(43) The organic acid in the raspberry fermented product is detected by adopting the agricultural industry standard NY/T 2277-2012 method of the People's Republic of China, and the results are shown in Table 3.

Example 4

(44) I. Preparation of a Culture Medium

(45) 1) Basic Culture Medium

(46) 10.0 g of casein peptone, 10.0 g of beef extract, 5.0 g of yeast powder, 5.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diammonium citrate, 1.0 g of tween 80, 2.0 g of K.sub.2HPO.sub.4, 0.2 g of MgSO.sub.4.Math.7H.sub.2O, 0.05 g of MnSO.sub.4.Math.H.sub.2O, and 10.0 g of CaCO.sub.3 are dissolved in 1 L of distilled water, and the pH value is adjusted to 6.8, to obtain a basic culture medium (20 g of agar is added when a solid culture medium is prepared).

(47) 2) Fermented Culture Medium

(48) pears are juiced to obtain a pear raw juice; distilled water is added to the pear raw juice for size mixing, and a volume ratio of the pear raw juice to distilled water is controlled to be 1:1 to obtain a pear juice.

(49) 10 g of sucrose, 4 g of ammonium nitrate, 1 g of manganese sulfate, 1 g of dipotassium hydrogen phosphate, and 10.0 g of CaCO.sub.3 are added to 1 L of the above pear juice, and the pH value is adjusted to about 6.8 to obtain a fermented culture medium.

(50) II. Fermentation

(51) 1) Activation of Strains

(52) Lactobacillus paracasei strains (CGMCC No. 14813) stored on an inclined plane are inoculated into the above solid basic culture medium, and then subjected to standing and culturing for about 24 hours under an anaerobic condition to obtain activated strains.

(53) 2) Preparation of a Seed Solution

(54) the above activated strains are inoculated into the above liquid basic culture medium, and then subjected to standing and culturing for about 24 h under the anaerobic condition to obtain a seed solution.

(55) 3) Fermentation

(56) according to the inoculation amount of 0.8% (v/v), the above seed solution is inoculated into the above fermented culture medium for fermenting and culturing, and the fermentation temperature is controlled to be at about 35? C., the rotation speed is controlled to be about 150 r/min, and the fermentation period is controlled to be about 7 days, to obtain a pear fermented product.

(57) The organic acid in the pear fermented product is detected by adopting the agricultural industry standard NY/T 2277-2012 method of the People's Republic of China, and the results are shown in Table 4.

Comparative Examples 1-4

(58) Lactobacillus paracasei standard strain ATCC 25302 is taken as a control in Comparative Examples 1-4, the rest are the same as those in Examples 1-4, except that Lactobacillus paracasei strain CGMCC No. 14813 in Examples 1-4 is replaced with the Lactobacillus paracasei standard strain ATCC 25302, and the inoculation amount is changed according to the following description.

(59) In Comparative Example 1, according to the inoculation amount of 3% (v/v), a seed solution is inoculated into a fermented culture medium for fermenting and culturing.

(60) In Comparative Example 2, according to the inoculation amount of 5% (v/v), a seed solution is inoculated into a fermented culture medium for fermenting and culturing.

(61) In Comparative Example 3, according to the inoculation amount of 1% (v/v), a seed solution is inoculated into a fermented culture medium for fermenting and culturing.

(62) In Comparative Example 4, according to the inoculation amount of 2% (v/v), a seed solution is inoculated into a fermented culture medium for fermenting and culturing.

(63) The contents of organic acids in fermented products of Comparative Example 1-4 are detected by the same method, and the detection results are shown in Tables 1 to 4 respectively.

(64) TABLE-US-00001 TABLE 1 Detection results of organic acids in grape fermented products of Example 1 and Comparative Example 1 Total content of the Organic acid (g/L) organic Lactic Acetic Propanoic Butyric Malic Tartaric Oxalic Citric acid Sample acid acid acid acid acid acid acid acid (g/L) Grape Juice 0.031 / 0.020 / 2.253 3.394 0.276 0.133 6.107 Example 1 18.342 0.512 0.135 0.009 0.093 1.669 0.942 0.103 21.805 Fermented product Comparative 12.395 0.490 0.067 0.025 0.037 0.301 0.071 0.103 13.489 Example 1 Fermented product Note: / means not detected.

(65) TABLE-US-00002 TABLE 2 Detection results of organic acids in blueberry fermented products of Example 2 and Comparative Example 2 Total content of the Organic acid (g/L) organic Lactic Acetic Propanoic Butyric Malic Tartaric Oxalic Citric acid Sample acid acid acid acid acid acid acid acid (g/L) Blueberry / / / / 0.192 0.055 0.145 3.134 3.526 juice Example 2 10.411 0.170 0.068 0.020 0.058 0.053 0.331 / 11.112 Fermented product Comparative 6.943 0.490 0.074 0.056 0.040 0.664 0.354 0.081 8.703 Example 2 Fermented product Note: / means not detected.

(66) TABLE-US-00003 TABLE 3 Detection results of organic acids in raspberry fermented products of Example 3 and Comparative Example 3 Total content of the Organic acid (g/L) organic Lactic Acetic Propanoic Butyric Malic Tartaric Oxalic Citric acid Sample acid acid acid acid acid acid acid acid (g/L) Raspberry / 0.224 / / 0.057 / 0.072 1.540 1.893 juice Example 3 3.250 0.939 0.160 0.035 0.388 1.430 0.487 0.119 6.807 Fermented product Comparative 2.048 0.166 0.352 0.026 0.062 0.355 0.169 0.152 3.330 Example 3 Fermented product Note: / means not detected.

(67) TABLE-US-00004 TABLE 4 Detection results of organic acids in pear fermented products of Example 4 and Comparative Example 4 Total content of the Organic acid (g/L) organic Lactic Acetic Propanoic Butyric Malic Tartaric Oxalic Citric acid Sample acid acid acid acid acid acid acid acid (g/L) Pear juice / / / / 1.33 0.057 0.217 1.041 2.645 Example 4 4.637 0.474 0.115 0.012 0.593 0.010 0.023 0.067 5.930 Fermented product Comparative 2.358 0.090 0.069 0.038 1.039 / 0.056 0.093 3.744 Example 4 Fermented product Note: / means not detected.

(68) The results in Tables 1 to 4 show that:

(69) compared with the unfermented raw juices and the fermented products fermented by Lactobacillus paracasei standard strain ATCC 25302, the content of the organic acid in the fermented products fermented by Lactobacillus paracasei (CGMCC No. 14813) of the present disclosure is significantly increased.

(70) Finally, it should be noted that the foregoing respective embodiments are merely intended for describing the technical solutions of the present disclosure other than limiting the present disclosure. Although the present disclosure is described in detail with reference to the foregoing respective embodiments, those ordinary skilled in the art should understand that the technical solutions described in the foregoing respective embodiments can still be modified, or some or all of the technical features can be equivalently substituted; however, these modifications or substitutions do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of respective embodiments of the present disclosure.