CN111334450A - Preparation process of lactobacillus lysis compound - Google Patents
Preparation process of lactobacillus lysis compound Download PDFInfo
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- CN111334450A CN111334450A CN201811552251.5A CN201811552251A CN111334450A CN 111334450 A CN111334450 A CN 111334450A CN 201811552251 A CN201811552251 A CN 201811552251A CN 111334450 A CN111334450 A CN 111334450A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/06—Lysis of microorganisms
- C12N1/066—Lysis of microorganisms by physical methods
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Abstract
A lactobacillus lysis compound preparation process relates to a microbial compound extraction technology, in particular to a lactobacillus lysis compound preparation process. A complete simplified process for the extraction of a lactic acid bacteria lysis complex is provided. Preparing a seed solution: respectively inoculating lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in MRS test tube culture medium, culturing for 22-26h at 37-38 ℃, then inoculating test tube strains into triangular flask liquid MRS culture medium for fermentation culture, and culturing for 35-38h at 37-38 ℃; fermentation: inoculating each seed solution into a fermentation tank according to the inoculation amount, and performing anaerobic fermentation; homogenizing and breaking the wall: homogenizing and breaking the wall of the fermented lactobacillus fermentation liquor by a high-pressure homogenizer to obtain a wall-broken liquid; centrifuging: passing the homogenized wall-breaking liquid through a tubular centrifuge to obtain centrifuged wall-breaking supernatant; and (3) filtering: and (3) passing the centrifuged wall-broken supernatant through a food-grade liquid filter to obtain the lactobacillus lysis compound. The method is simple and convenient, the process is simplified, and the operation is easy.
Description
Technical Field
The invention relates to a microbial compound extraction technology, in particular to a lactobacillus lysis compound preparation technology.
Background
Lactic acid bacteria or fermented biological products thereof are thought to have a function of improving the micro-ecological environment of the intestinal tract of a human body by regulating the functions of the immune system of a host and improving the balance of the flora of the host to produce beneficial physiological effects on the host. The functions of the utility model are as follows: (1) improving the intestinal micro-ecological environment; (2) the immunity is improved; (3) the bowel is unblocked, the stool is moistened, and the intestinal function is improved; (4) inhibiting endotoxin production, and delaying aging. The intracellular product of the lactic acid bacteria is used as a novel biological resource, and the research and development of industrialization of the intracellular product of the lactic acid bacteria bring more positive significance to daily chemicals and food. However, the production and preparation process of the lactobacillus lysate is complicated, and a complete and simplified production process is required to obtain a high-quality lactobacillus lysis complex.
Disclosure of Invention
The present invention addresses the above problems by providing a complete and simplified process for the extraction of a complex of lactic acid bacteria lysis.
The technical scheme of the invention is as follows: the method comprises the following steps:
1) and preparing seed liquid: respectively inoculating lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in MRS test tube culture medium, culturing for 22-26h at 37-38 ℃, then inoculating test tube strains into triangular flask liquid MRS culture medium for fermentation culture, and culturing for 35-38h at 37-38 ℃;
2) and fermenting: inoculating each seed solution into a fermentation tank according to the inoculation amount, and performing anaerobic fermentation at the fermentation temperature of 37-38 ℃;
3) homogenizing and breaking the wall: homogenizing and breaking the wall of the fermented lactobacillus fermentation liquor by a high-pressure homogenizer to obtain a wall-broken liquid;
4) and centrifuging: centrifuging the homogenized wall-breaking liquid at a high speed of 8000rpm/min by a tubular centrifuge to remove thallus residues to obtain centrifuged wall-breaking supernatant;
5) and filtering: and (3) passing the centrifuged wall-broken supernatant through a food-grade liquid filter to obtain the lactobacillus lysis compound.
The inoculation amounts of the lactobacillus plantarum, the bifidobacterium longum and the lactobacillus paracasei in the step 2) are 3-4%, 5-6% and 3-4% respectively.
The conditions for homogenizing and breaking the wall in the step 3) are as follows: the homogeneous wall-breaking pressure is 120-130bar, and the wall breaking is carried out twice.
In the step 5), a fiber membrane with the diameter of 0.15-0.20 μm is adopted for filtration.
The invention has the beneficial effects that: the lactobacillus lysate is extracted by simple processes such as homogenizing wall breaking, centrifuging, filtering and the like, and the method is more suitable for high-efficiency preparation of the lactobacillus lysate, simple and convenient, simplified in process and easy to operate.
Detailed Description
The present invention will be described in detail with reference to examples.
Example 1
The invention discloses a preparation process of a lactobacillus lysis compound, which comprises the following steps:
1) and preparing seed liquid: respectively inoculating lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in MRS test tube culture medium, culturing for 22h at 37 ℃, then inoculating test tube strains into triangular flask liquid MRS culture medium for fermentation culture, and culturing for 35h at 37 ℃;
2) and fermenting: inoculating each seed solution into a fermentation tank according to the inoculation amount, and performing anaerobic fermentation at the fermentation temperature of 37 ℃; the inoculation amounts of lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei are respectively 3%, 5% and 3%;
3) homogenizing and breaking the wall: homogenizing and breaking the wall of the fermented lactobacillus fermentation liquor by a high-pressure homogenizer to obtain a wall-broken liquid; the conditions for homogenizing and breaking the wall are as follows: homogenizing at 120bar for wall breaking, and performing twice homogenizing for wall breaking;
4) and centrifuging: centrifuging the homogenized wall-breaking liquid at a high speed of 8000rpm/min by a tubular centrifuge to remove thallus residues to obtain centrifuged wall-breaking supernatant;
5) and filtering: passing the centrifuged cell wall-broken supernatant through a food-grade liquid filter, such as a 0.15 μm fiber membrane, to obtain a lactobacillus lysis complex.
Example 2
The invention discloses a preparation process of a lactobacillus lysis compound, which comprises the following steps:
1) and preparing seed liquid: respectively inoculating lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in MRS test tube culture medium, culturing for 26h at 38 ℃, then inoculating test tube strains into triangular flask liquid MRS culture medium for fermentation culture, and culturing for 38h at 38 ℃;
2) and fermenting: inoculating each seed solution into a fermentation tank according to the inoculation amount, and performing anaerobic fermentation at the fermentation temperature of 38 ℃; the inoculation amounts of lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei are respectively 4%, 6% and 4%;
3) homogenizing and breaking the wall: homogenizing and breaking the wall of the fermented lactobacillus fermentation liquor by a high-pressure homogenizer to obtain a wall-broken liquid; the conditions for homogenizing and breaking the wall are as follows: homogenizing and breaking wall at 130bar, and performing homogenization and wall breaking twice;
4) and centrifuging: centrifuging the homogenized wall-breaking liquid at a high speed of 8000rpm/min by a tubular centrifuge to remove thallus residues to obtain centrifuged wall-breaking supernatant;
5) and filtering: passing the centrifuged cell wall-broken supernatant through a food-grade liquid filter, such as a 0.20 μm fiber membrane, to obtain a lactobacillus lysis complex.
Example 3
The invention discloses a preparation process of a lactobacillus lysis compound, which comprises the following steps:
1) and preparing seed liquid: respectively inoculating lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in MRS test tube culture media, culturing for 24h at 37 ℃, then inoculating test tube strains into a triangular flask liquid MRS culture media for fermentation culture, and culturing for 36h at 37 ℃;
2) and fermenting: inoculating each seed solution into a fermentation tank according to the inoculation amount, and performing anaerobic fermentation at the fermentation temperature of 37 ℃; the inoculation amounts of lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei are respectively 3%, 5% and 3%;
3) homogenizing and breaking the wall: homogenizing and breaking the wall of the fermented lactobacillus fermentation liquor by a high-pressure homogenizer to obtain a wall-broken liquid; the conditions for homogenizing and breaking the wall are as follows: homogenizing at 120bar for wall breaking, and performing twice homogenizing for wall breaking;
4) and centrifuging: centrifuging the homogenized wall-breaking liquid at a high speed of 8000rpm/min by a tubular centrifuge to remove thallus residues to obtain centrifuged wall-breaking supernatant;
5) and filtering: passing the centrifuged cell wall-broken supernatant through a food-grade liquid filter, such as a 0.15 μm fiber membrane, to obtain a lactobacillus lysis complex.
Claims (4)
1. A process for preparing a complex for lysing lactic acid bacteria, comprising the steps of:
1) and preparing seed liquid: respectively inoculating lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei in MRS test tube culture medium, culturing for 22-26h at 37-38 ℃, then inoculating test tube strains into triangular flask liquid MRS culture medium for fermentation culture, and culturing for 35-38h at 37-38 ℃;
2) and fermenting: inoculating each seed solution into a fermentation tank according to the inoculation amount, and performing anaerobic fermentation at the fermentation temperature of 37-38 ℃;
3) homogenizing and breaking the wall: homogenizing and breaking the wall of the fermented lactobacillus fermentation liquor by a high-pressure homogenizer to obtain a wall-broken liquid;
4) and centrifuging: centrifuging the homogenized wall-breaking liquid at a high speed of 8000rpm/min by a tubular centrifuge to remove thallus residues to obtain centrifuged wall-breaking supernatant;
5) and filtering: and (3) passing the centrifuged wall-broken supernatant through a food-grade liquid filter to obtain the lactobacillus lysis compound.
2. The process of claim 1, wherein the amounts of Lactobacillus plantarum, Bifidobacterium longum, and Lactobacillus paracasei inoculated in step 2) are 3-4%, 5-6%, and 3-4%, respectively.
3. The process of claim 1, wherein the conditions for breaking the cell wall in step 3) are: the homogeneous wall-breaking pressure is 120-130bar, and the wall breaking is carried out twice.
4. The process of claim 1, wherein said step 5) is performed by filtration using a 0.15-0.20 μm fiber membrane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811552251.5A CN111334450A (en) | 2018-12-19 | 2018-12-19 | Preparation process of lactobacillus lysis compound |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811552251.5A CN111334450A (en) | 2018-12-19 | 2018-12-19 | Preparation process of lactobacillus lysis compound |
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| CN111334450A true CN111334450A (en) | 2020-06-26 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113632922A (en) * | 2021-08-16 | 2021-11-12 | 科郦有限公司 | Lysate, preparation method and application |
| CN113755365A (en) * | 2021-08-11 | 2021-12-07 | 天津科技大学 | Preparation method and application of lactobacillus extract |
| CN114601174A (en) * | 2020-12-04 | 2022-06-10 | 安琪纽特股份有限公司 | Lactobacillus lysate and preparation method and application thereof |
| CN115595333A (en) * | 2022-12-15 | 2023-01-13 | 朗肽生物制药股份有限公司（Cn） | Composite lactobacillus fermentation lysate and preparation method and application thereof |
-
2018
- 2018-12-19 CN CN201811552251.5A patent/CN111334450A/en not_active Withdrawn
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114601174A (en) * | 2020-12-04 | 2022-06-10 | 安琪纽特股份有限公司 | Lactobacillus lysate and preparation method and application thereof |
| CN114601174B (en) * | 2020-12-04 | 2024-06-25 | 安琪纽特股份有限公司 | Lactic acid bacteria lysate and preparation method and application thereof |
| CN113755365A (en) * | 2021-08-11 | 2021-12-07 | 天津科技大学 | Preparation method and application of lactobacillus extract |
| CN113755365B (en) * | 2021-08-11 | 2024-02-27 | 天津科技大学 | Preparation method and application of lactobacillus extract |
| CN113632922A (en) * | 2021-08-16 | 2021-11-12 | 科郦有限公司 | Lysate, preparation method and application |
| CN115595333A (en) * | 2022-12-15 | 2023-01-13 | 朗肽生物制药股份有限公司（Cn） | Composite lactobacillus fermentation lysate and preparation method and application thereof |
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Application publication date: 20200626 |