CN112941004A

CN112941004A - Formula of growth factor for promoting proliferation of lactic acid bacteria and application method thereof

Info

Publication number: CN112941004A
Application number: CN202110494299.0A
Authority: CN
Inventors: 周群; 王永平; 王维新; 周亚男; 吕晋江
Original assignee: Weikaihaisi Shandong Bioengineering Co ltd
Current assignee: Weikaihaisi Shandong Bioengineering Co ltd
Priority date: 2021-05-07
Filing date: 2021-05-07
Publication date: 2021-06-11

Abstract

The invention discloses a formula of a growth factor for promoting lactobacillus proliferation and an application method thereof, wherein the formula comprises the following components in parts by weight: the growth factor solution comprises the following components in parts by mass: 10% of ascorbic acid solution and 10% of biotin solution; 10% of a tyrosine solution; 10% of galacto-oligosaccharide solution; filtering the solution to remove bacteria for later use; the method specifically comprises the following steps: selecting the final concentration of ascorbic acid to be 0.1g/L, the final concentration of biotin to be 0.2g/L, the final concentration of tyrosine to be 0.1g/L and the concentration of galacto-oligosaccharide to be added; the invention adopts a mode of quantitatively adding the growth factors to replace the original mode of utilizing plant juice or hydrolysate thereof as the growth factors to realize the high-density culture of the lactic acid bacteria.

Description

Formula of growth factor for promoting proliferation of lactic acid bacteria and application method thereof

Technical Field

The invention relates to a formula for promoting lactobacillus proliferation, in particular to a formula of a growth factor for promoting lactobacillus proliferation and an application method thereof, and belongs to the technical field of promoting lactobacillus proliferation.

Background

In the prior art, the problem that the lactobacillus culture medium is lack of growth factors is generally supplemented by plant juice and hydrolysate thereof (such as tomato juice, kelp juice, compound fruit juice, bean pulp extracting solution, bran hydrolysate, soybean hydrolysate and the like); however, the utilization of plant juice and its hydrolysate as a growth factor has the following disadvantages:

1. complex operation and large investment: the method of using plant juice and its hydrolysate as growth factor needs additional equipment for cleaning, crushing, leaching or hydrolyzing, filtering, centrifuging and other operations of the required plant, and this results in long production flow line, increased production waste, complicated production process, high labor strength, great equipment investment in the early stage, etc.

2. The quality of the raw materials is difficult to control: the quality of plants varies with the place of production, the variety and the like, and is influenced by seasons, the supply of plant raw materials is also influenced to a certain extent, and the instability of the quality and the supply of the raw materials can possibly cause the instability of the production and the product quality; furthermore, the price of the plants is also affected by the market price, so that the production costs become uncontrollable; and the current commercial plant extract finished products are mainly used for medicine and health care, and if the plant extract finished products are used as main raw materials for production, the production cost is further increased.

3. Large dosage and difficult storage: the method of using plant juice and hydrolysate thereof as growth factors requires a relatively large amount of plants, which not only makes the production process complicated and complicated, but also needs to reserve a certain amount of raw materials in a warehouse in order to ensure the continuity of the production, and the raw materials inevitably go bad during storage, which not only causes the waste of raw materials, but also may affect the subsequent process due to improper treatment, even the final product quality, and other problems.

Disclosure of Invention

In order to solve the problems, the invention provides a formula of a growth factor for promoting the proliferation of lactic acid bacteria and an application method thereof, the high-density culture of the lactic acid bacteria is realized by adding the growth factor, and a large amount of active thalli of the lactic acid bacteria are obtained after the fermentation is finished.

The formula of the growth factor for promoting lactobacillus proliferation comprises the following components:

comprises MRS culture medium and growth factor solution; the MRS culture medium comprises the following components in parts by mass: 10g of peptone, 5g of beef powder, 4g of yeast powder and K₂HPO₄•7H₂O2 g, triammonium citrate 2g, sodium acetate 3H₂O5 g, glucose 20g, Tween 801mL, MgSO₄•7H₂O 0.2g；MnSO₄•4H₂0.05g of O and 1000mL of distilled water;

the growth factor solution comprises the following components in parts by mass: 10% of ascorbic acid solution and 10% of biotin solution; 10% of a tyrosine solution; 10% of galacto-oligosaccharide solution; and filtering the solution to remove bacteria for later use.

A growth factor application method for promoting lactobacillus proliferation comprises selecting ascorbic acid final concentration of 0.1g/L, biotin final concentration of 0.2g/L, tyrosine 0.1g/L and galactooligosaccharide concentration of 0.3g/L for adding.

Compared with the prior art, the formula of the growth factor for promoting lactobacillus proliferation and the application method thereof have the following beneficial effects:

the method adopts a mode of quantitatively adding the growth factors to replace the original mode of utilizing plant juice or hydrolysate thereof as the growth factors, so that the high-density culture of the lactic acid bacteria is realized; through the change, the yield of the lactic acid bacteria is improved, the production flow is simplified, the equipment investment is reduced, the labor intensity is reduced, the raw materials are easy to store, the quality is stable, the original high-pressure steam sterilization is replaced by a filter membrane filtration mode, the energy consumption is reduced, and the damage of growth factors caused by high-pressure steam sterilization is avoided.

Drawings

FIG. 1 is a schematic diagram of the test results of a comparative experiment of the present invention.

Detailed Description

comprises MRS culture medium and growth factor solution; the MRS culture medium comprises the following components in parts by mass: 10g of peptone, 5g of beef powder, 4g of yeast powder and K₂HPO₄•7H₂O2 g, triammonium citrate 2g, sodium acetate 3H₂O5 g, glucose 20g, Tween 801mL, MgSO₄•7H₂O 0.2g；MnSO₄•4H₂0.05g of O and 1000mL of distilled water; the growth factor solution comprises the following components in parts by mass: 10% of ascorbic acid solution and 10% of biotin solution; 10% of a tyrosine solution; 10% of galacto-oligosaccharide solution; filtering the solution to remove bacteria for later use;

a growth factor application method for promoting lactobacillus proliferation selects ascorbic acid final concentration of 0.1g/L, biotin final concentration of 0.2g/L, tyrosine of 0.1g/L and galacto-oligosaccharide of 0.3g/L to be added, and as the test result, the number of the lactic acid bacteria shows a descending trend along with the increase of the addition of the ascorbic acid, and the number of the lactic acid bacteria shows a trend of firstly ascending and then descending along with the increase of the addition of the biotin; along with the increase of the addition amount of tyrosine, the number of the lactic acid bacteria is not changed greatly; and along with the increase of the addition amount of the galacto-oligosaccharide, the number of the lactic acid bacteria presents an obvious rising trend, so that the increment point of the components with the trend of falling points is kept, and the total amount of the components is balanced by using the addition amount of the galacto-oligosaccharide, so that the growth factors achieve mutual synergistic cooperation under the condition of low dosage, and the high-density culture of the lactic acid bacteria is realized.

Example 1:

respectively adding 300mL of MRS culture medium into 5 conical flasks with the volume of 500mL, sterilizing and cooling, and respectively adding 0.3mL of sterile 10% ascorbic acid solution and 1.8mL of sterile water; 0.3mL of 10% ascorbic acid solution, 0.6mL of 10% biotin solution and 1.2mL of sterile water; 0.3mL of 10% ascorbic acid solution, 0.6mL of 10% biotin solution, 0.3mL of 10% tyrosine solution and 0.9mL of sterile water; 0.3mL of 10% ascorbic acid solution, 0.6mL of 10% biotin solution, 0.3mL of 10% tyrosine solution and 0.9mL of 10% galacto-oligosaccharide solution; forming three gradients of 0.1g/L, 0.2g/L and 0.3g/L in the final concentration of different growth factors in a culture medium, adding 2.1mL of sterile water as a blank control, respectively inoculating the prepared seed liquid into the culture medium according to the inoculation amount of 1%, culturing at 37 ℃ for 24h, and detecting the viable count of lactic acid bacteria in the fermentation liquid in a coating counting mode after fermentation is finished; the fermentation results are shown in FIG. 1; as can be seen from fig. 1, with the increase of the types of the growth factors in the fermentation broth, the viable count of the lactic acid bacteria in the fermentation broth is greatly increased, which indicates that the four growth factors do not exhibit an antagonistic effect, and the counting result of the viable count of the lactic acid bacteria indicates that the four growth factors exhibit a synergistic effect, thereby jointly promoting the growth of the lactic acid bacteria.

The formula of the growth factor for promoting the proliferation of the lactic acid bacteria utilizes a 100L fermentation tank of the factory area to carry out a control experiment, and the experiment is divided into: no growth factor and plant juice are added, only plant juice is added, and only growth factor is added, except the three variables, the rest materials and the fermentation control method are consistent with the production; test results show that in three groups of tests that no growth factor and plant juice are added into the fermentation liquor, only plant juice is added into the fermentation liquor, and only growth factor is added into the fermentation liquor, the counting results of the viable count of the lactobacillus in the fermentation liquor are respectively 4 multiplied by 109CFU/mL, 8.5 multiplied by 109CFU/mL and 10 multiplied by 109CFU/mL when the fermentation is finished; the results show that the formula of the growth factor not only can completely replace plant juice, but also has better fermentation effect than the plant juice; in addition, the cost accounting proves that the production cost is reduced, the production flow is simplified and the production efficiency is improved by replacing the plant juice with the growth factor.

The above-described embodiments are merely preferred embodiments of the present invention, and all equivalent changes or modifications of the structures, features and principles described in the claims of the present invention are included in the scope of the present invention.

Claims

1. The formula of the growth factor for promoting the proliferation of the lactic acid bacteria is characterized by comprising the following components in percentage by weight:

2. A method for applying a growth factor for promoting the proliferation of lactic acid bacteria comprises the following specific steps: the final concentration of ascorbic acid is 0.1g/L, the final concentration of biotin is 0.2g/L, tyrosine is 0.1g/L and galacto-oligosaccharide is 0.3 g/L.