CN111662841A - Method for preparing efficient lactic acid bacteria culture medium by liquid fermentation of edible hyphae - Google Patents
Method for preparing efficient lactic acid bacteria culture medium by liquid fermentation of edible hyphae Download PDFInfo
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- CN111662841A CN111662841A CN202010521879.XA CN202010521879A CN111662841A CN 111662841 A CN111662841 A CN 111662841A CN 202010521879 A CN202010521879 A CN 202010521879A CN 111662841 A CN111662841 A CN 111662841A
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Abstract
The invention discloses a method for preparing a high-efficiency lactic acid bacteria culture medium by liquid fermentation of edible mycelia, which comprises the steps of grinding a proper amount of liquid fermentation edible mycelia into slurry, adding water, boiling, filtering, fixing the volume, adjusting the pH value to obtain a filtrate culture medium, subpackaging and sterilizing to obtain the lactic acid bacteria culture medium. The liquid culture lactic acid bacteria culture medium prepared by the invention has low cost and simple preparation, and the abundant protein and free nucleotide in the mycelium can effectively reduce the death number of lactic acid bacteria, delay the death time and avoid the degeneration and variation of the thallus; compared with the common MRS culture medium, the OD600 value after 18h of culture is obviously superior to that of the MRS culture medium, and the OD600 value of the finally optimized culture medium is more than 1.05 times of that of the common MRS culture medium.
Description
Technical Field
The invention relates to a method for preparing a high-efficiency liquid culture medium for lactic acid bacteria by using edible hyphae.
Background
Lactic acid bacteria are gram-positive bacteria which can improve the animal's immunity, digestion and metabolism and have strict nutritional requirements. When the lactic acid bacteria culture medium is selected, the growth and the rapid propagation of the lactic acid bacteria are facilitated, and the stress resistance of the bacterial cells can be properly improved. The culture medium used in industrial production has low cost, easily available raw materials and high thallus yield. Although there are many types of lactic acid bacteria culture media, basic culture media can be roughly classified into 4 types: whey protein culture medium, skim milk emulsion zymolyte culture medium, skim milk culture medium and MRS emulsion zymolyte culture medium. The MRS culture medium is suitable for the growth of various lactobacilli, but the growth speed of the lactobacilli in the culture medium is low, the formula is complex, the cost is high, and the MRS culture medium is not suitable for being applied to food production due to poor taste.
The skim milk culture medium is a culture medium mainly used in domestic lactic acid bacteria industrial production, and although the skim milk culture medium is low in cost and economic and easily available in raw materials, the skim milk culture medium is high in viscosity and not beneficial to concentration and separation of lactic acid bacteria. The whey protein culture medium is a culture medium mainly used in the foreign lactobacillus industrial production, but whey protein in the whey protein culture medium is easy to generate thermal denaturation and precipitate, so that the viable count of the lactobacillus is low.
At present, most of domestic culture mediums for propagating lactic acid bacteria are improved on the basis of classical MRS culture mediums, and the cost of the culture mediums can account for more than 10% of the production cost due to the fact that most of the culture mediums are added with expensive components such as yeast powder, beef extract and the like. The traditional MRS culture medium has complex components and high cost for culturing lactic acid bacteria, so that the problem of economic cost is faced in large-scale industrial culture of lactic acid bacteria. Many studies have optimized and adapted MRS media in order to reduce costs, meet production needs or increase target yields. Therefore, how to optimize the formula of the lactic acid bacteria culture medium, reduce the cost and improve the viable count and the lactic acid yield becomes a hotspot of the research of the lactic acid bacteria culture medium.
Disclosure of Invention
The invention aims to solve the technical problems of complex components of MRS culture medium and high cost of culturing lactic acid bacteria in the prior art, and provides a method for preparing a high-efficiency liquid culture medium for culturing lactic acid bacteria by using edible hypha.
In order to solve the technical problems, the invention provides the following technical scheme:
a method for preparing a high-efficiency liquid culture medium for lactic acid bacteria by using edible hypha comprises the following steps:
s1, weighing a proper amount of edible mycelia, grinding the edible mycelia into slurry, adding a proper amount of water into the mycelium slurry, boiling for 15-25 minutes, filtering by using two layers of medical gauze, and fixing the final volume of the filtrate to a proper volume by supplementing water;
s2, adjusting the pH value to 6.2-6.8 to obtain a filtrate culture medium, subpackaging the filtrate culture medium in 50mL conical flasks, and sterilizing at 121 ℃ for 20 min;
s3, inoculating the lactobacillus seed liquid into the filtrate culture medium, and culturing at 37 ℃.
Further, the edible mycelium in S1 is liquid fermented shiitake mushroom mycelium, water is added into a wall breaking machine, the liquid fermented shiitake mushroom mycelium is ground at high speed to prepare mycelium slurry, and the lactobacillus in S3 is lactobacillus plantarum LP-G18.
Further, the amount of the shiitake mushroom mycelia was 225g/1000mL of the filtrate medium.
Further, in S1, 4-6 g/L of sodium acetate and 0.4-0.6 mg/L of riboflavin are added before constant volume.
Further, the edible mycelium in S1 is Pleurotus eryngii mycelium, water is added into a stirrer to grind at high speed to prepare mycelium slurry, and the lactic acid bacteria in S3 is rhamnose lactic acid bacteria.
Furthermore, 1.5-2.5 g/L, MgSO 4.7H 2O 0.1.1-0.3 mg/L of ammonium citrate is added into S1 before constant volume.
The comparison document 2015107987146 discloses a low-cost high-efficiency lactobacillus culture medium and application thereof, and discloses a method for adding pleurotus eryngii into a lactobacillus culture medium, but the added pleurotus eryngii has fruiting bodies, the mycelium of edible fungi has high polysaccharide content, and the mycelium of edible fungi also contains rich protein and free amino acids, especially amino acids required by growth of lactic acid bacteria such as aspartic acid, glutamic acid, arginine and the like, so that good nutritional ingredients are provided, the fermentation period of the mycelium is short, the investment cost is low, the production operation is simple, the yield is high, the method is easy to control, the method is more suitable for industrial production and the like, and the defects of long cultivation period, large manual investment, easy limitation of climatic conditions, heavy metal enrichment pollution and the like caused by the fruiting bodies can be alleviated. Therefore, the corresponding culture medium is prepared by utilizing the mycelium produced by liquid fermentation of the edible fungi, so that the survival conditions of the lactic acid bacteria are improved.
The invention has the following beneficial effects: the liquid culture lactic acid bacteria culture medium prepared by the invention has low cost and simple preparation, and the abundant protein and free nucleotide in the mycelium can effectively reduce the death number of lactic acid bacteria, delay the death time and avoid the degeneration and variation of the thallus; compared with the common MRS culture medium, the OD600 value after 18h of culture is obviously superior to that of the MRS culture medium, and the OD600 value of the finally optimized culture medium is more than 1.05 times of that of the common MRS culture medium.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 is a comparison of the medium of example 1 with MRS medium;
FIG. 2 is a comparison of the medium of example 2 with MRS medium.
Detailed Description
The preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.
Example 1
A method for preparing a high-efficiency lactic acid bacteria culture medium by liquid fermentation of edible hyphae comprises the following steps:
s1, weighing 225g of appropriate amount of liquid fermented shiitake mushroom mycelia, adding water in a wall breaking machine, grinding at high speed to prepare mycelium slurry, adding appropriate amount of water in the mycelium slurry, boiling for 15-25 minutes, filtering with two layers of medical gauze, adding 5g of sodium acetate and 0.6mg of riboflavin, and adding water to fix the final volume of the filtrate to 1000 mL;
s2, adjusting the pH value to 6.2-6.4 to obtain a filtrate culture medium, subpackaging the filtrate culture medium in 50mL conical flasks, and sterilizing at 121 ℃ for 20 min;
s3, inoculating the seed liquid of the lactobacillus plantarum LP-G18 into the filtrate culture medium, and culturing at 37 ℃.
The mycelium of the pleurotus eryngii contains rich protein and free amino acid, particularly aspartic acid, glutamic acid and arginine, the OD value of the culture medium is 1.05 times of that of a common MRS culture medium, and the survival rate of lactic acid bacteria in the optimized culture medium at the later stage of culture is higher.
Example 2
A method for preparing a high-efficiency lactic acid bacteria culture medium by liquid fermentation of edible hyphae comprises the following steps:
s1, weighing 200g of liquid fermented Lentinus Edodes mycelium, adding water, grinding at high speed to obtain mycelium slurry, adding appropriate amount of water into the mycelium slurry, boiling for 15-25 min, filtering with two layers of medical gauze, adding 1.5g/L ammonium citrate and MgSO4·7H2O0.3 mg, and the final volume of the filtrate is fixed to 1000mL by supplementing water;
s2, adjusting the pH value to 6.8 to obtain a filtrate culture medium, subpackaging the filtrate culture medium in 50mL conical flasks, and sterilizing at 121 ℃ for 20 min;
s3, inoculating the rhamnose lactobacillus seed liquid into the filtrate culture medium, and culturing at 37 ℃.
The OD value of the culture medium of the embodiment is 1.1 times of that of a common MRS culture medium, the survival rate of lactic acid bacteria in the optimized culture medium at the later stage of culture is higher, and the culture medium is an excellent lactic acid bacteria culture medium and is suitable for industrial large-scale production.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A method for preparing a high-efficiency lactic acid bacteria culture medium by liquid fermentation of edible hyphae is characterized by comprising the following steps:
s1, weighing a proper amount of liquid fermented edible mycelia, grinding the edible mycelia into slurry, adding a proper amount of water into the mycelium slurry, boiling for 15-25 minutes, filtering by using two layers of medical gauze, and fixing the final volume of the filtrate to a proper volume by supplementing water;
s2, adjusting the pH value to 6.2-6.8 to obtain a filtrate culture medium, subpackaging the filtrate culture medium in 50mL conical flasks, and sterilizing at 121 ℃ for 20 min;
s3, inoculating the lactobacillus seed liquid into the filtrate culture medium, and culturing at 37 ℃.
2. The method for preparing culture medium for high-efficiency lactic acid bacteria from liquid fermented edible hyphae according to claim 1, wherein the edible hyphae in S1 is liquid fermented Lentinus edodes mycelium, and is ground into hyphae slurry by adding water in a wall breaking machine at high speed, and the lactic acid bacteria in S3 is Lactobacillus plantarum LP-G18.
3. The method for preparing culture medium for Lactobacillus plantarum according to claim 2, wherein the amount of mycelia of Lentinus edodes was 225g/1000mL of the filtrate medium.
4. The method for preparing a high-efficiency lactic acid bacteria culture medium by liquid fermentation of edible hyphae as claimed in claim 2 or 3, wherein 4-6 g/L of sodium acetate and 0.4-0.6 mg/L of riboflavin are added before volume fixing in S1.
5. The method for preparing a culture medium for highly effective lactic acid bacteria from liquid fermented edible hyphae according to claim 1, wherein the edible hyphae in S1 is liquid fermented Pleurotus eryngii mycelium, and is ground into hypha slurry with water in a mixer at high speed, and the lactic acid bacteria in S3 is rhamnose lactic acid bacteria.
6. The method for preparing a high-efficiency lactic acid bacteria culture medium by liquid fermentation edible hyphae according to claim 1, wherein 1.5-2.5 g/L, MgSO 4-7H 2O 0.1.1-0.3 mg/L of ammonium citrate is added before volume measurement in S1.
7. The method for preparing a culture medium for high-efficiency lactic acid bacteria by liquid fermentation of edible hyphae according to claim 1, wherein the dosage of the Pleurotus eryngii mycelium in S1 is 200g/1000mL of the filtrate culture medium.
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| CN105349458A (en) * | 2015-11-19 | 2016-02-24 | 江苏省农业科学院 | Low-cost high-efficiency lactic acid bacteria medium and uses thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105349458A (en) * | 2015-11-19 | 2016-02-24 | 江苏省农业科学院 | Low-cost high-efficiency lactic acid bacteria medium and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| 梁宝东等: "香菇菌丝体酸奶生产工艺的研究", 《食品科技》 * |
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