CN112358989A - Lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage and preparation method of lactic acid bacteria agent - Google Patents
Lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage and preparation method of lactic acid bacteria agent Download PDFInfo
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- 239000004310 lactic acid Substances 0.000 title claims abstract description 47
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 47
- 241000894006 Bacteria Species 0.000 title claims abstract description 46
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- 238000002360 preparation method Methods 0.000 title claims abstract description 23
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- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 18
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims abstract description 17
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- 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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- A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
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Abstract
A lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage comprises the following components in parts by weight: 2-3 parts of sodium acetate, 10-20 parts of glucose or maltose, 0.15-3 parts of aspartic acid, 5-10 parts of beef extract, 1.5-2 parts of calcium carbonate, 15-20 parts of calcium hydroxide, 100-200 parts of tomato juice, 3-5 parts of inorganic salt, 0.5-1 part of tryptone and 2-5 parts of yeast extract. According to the industrial lactic acid bacteria culture solution for the pickled Chinese cabbage without the salt and the preparation method of the lactic acid bacteria agent, the preparation raw material cost of the culture solution is low, the culture solution is easy to obtain, and the fermentation period of the pickled Chinese cabbage without the salt can be shortened; compared with a direct-vat-set lactic acid bacteria agent, the preparation process of the lactic acid bacteria agent is simpler and more convenient, the cost is lower, the control is convenient, and the lactic acid bacteria agent can be suitable for most industrial production.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage and a preparation method of a lactic acid bacteria agent.
Background
The fermentation process of the pickled Chinese cabbage is accompanied with the growth and metabolism activities of microorganisms mainly containing lactic acid bacteria, but the traditional natural fermentation process has long period and unstable product quality, so the artificial inoculation lactic acid bacteria fermentation pickled Chinese cabbage technology gradually replaces the natural fermentation production technology. Compared with the traditional natural fermentation production technology, the fermentation period of the pickled Chinese cabbage by artificially inoculating the lactobacillus for fermentation is shorter, the safety is higher, and the product quality is more stable. The technology of artificially inoculating lactobacillus to ferment the pickled Chinese cabbage becomes a hot spot of the current research. With the gradual rise of the salt-free pickled Chinese cabbage, the production mode of artificially inoculating lactobacillus to the low-salinity vegetable to strengthen fermentation replaces the natural fermentation production mode, and gradually becomes the development trend of most pickled Chinese cabbage enterprises.
In recent years, a direct vat set lactobacillus preparation is gradually applied to the fermentation production of the pickled Chinese cabbage without the salt, and the direct vat set lactobacillus preparation has the advantages of simplicity and convenience in use, long preservation time, good product quality and the like, but the preparation process is complicated and high in cost, a large scale cannot be formed in the industry of the pickled Chinese cabbage without the salt, the market price of the common pickled Chinese cabbage without the salt is relatively low, the preparation cost is not high, and the preparation cost of the pickled Chinese cabbage without the salt is increased due to the use of the direct vat set lactobacillus preparation, so that the direct vat set lactobacillus preparation is not suitable for most industrial production, and the actual application is not wide.
Disclosure of Invention
In order to solve the defects in the prior art, the invention provides the industrial lactobacillus culture solution for the pickled Chinese cabbage without the salt and the preparation method of the lactobacillus agent.
In order to achieve the purpose, the invention adopts the specific scheme that: a lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage comprises the following components in parts by weight: 2-3 parts of sodium acetate, 10-20 parts of glucose or maltose, 0.15-3 parts of aspartic acid, 5-10 parts of beef extract, 1.5-2 parts of calcium carbonate, 15-20 parts of calcium hydroxide, 100-200 parts of tomato juice, 3-5 parts of inorganic salt, 0.5-1 part of tryptone and 2-5 parts of yeast extract.
The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage is further optimized: also comprises 20-25 parts of beer liquid.
The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage is further optimized: the composite material comprises the following components in parts by mass: 3 parts of sodium acetate, 10 parts of glucose or maltose, 0.2 part of aspartic acid, 10 parts of beef extract, 16 parts of calcium carbonate, 15 parts of calcium hydroxide, 150 parts of tomato juice, 3 parts of inorganic salt, 1 part of tryptone and 3 parts of yeast extract.
The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage is further optimized: the composite material comprises the following components in parts by mass: 2 parts of sodium acetate, 10 parts of glucose or maltose, 0.15 part of aspartic acid, 8 parts of beef extract, 20 parts of calcium carbonate, 15 parts of calcium hydroxide, 200 parts of tomato juice, 5 parts of inorganic salt, 1 part of tryptone, 2 parts of yeast extract and 20 parts of beer liquor.
The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage is further optimized: the composite material comprises the following components in parts by mass: 2.5 parts of sodium acetate, 10 parts of glucose or maltose, 0.3 part of aspartic acid, 10 parts of beef extract, 15 parts of calcium carbonate, 15 parts of calcium hydroxide, 200 parts of tomato juice, 3 parts of inorganic salt, 0.8 part of tryptone, 5 parts of yeast extract and 25 parts of beer liquor.
The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage is further optimized: the composite material comprises the following components in parts by mass: 2 parts of sodium acetate, 10 parts of glucose or maltose, 0.15 part of aspartic acid, 10 parts of beef extract, 20 parts of calcium carbonate, 20 parts of calcium hydroxide, 100 parts of tomato juice, 3 parts of inorganic salt, 1 part of tryptone, 3 parts of yeast extract and 20 parts of beer liquor.
A preparation method of a lactic acid bacteria agent for industrial non-salt pickled Chinese cabbage comprises the following steps:
s1, strain activation: inoculating the preserved strain into an MRS culture medium, and carrying out anaerobic culture at 37 ℃ for 24h to obtain an activated strain;
s2, activated strain culture: placing the activated strain into a bottle filled with the culture solution of any one of claims 1 to 6, performing shake culture at 32 to 40 ℃ for 12 to 24 hours, measuring the pH value every 4 hours, adjusting the pH value to 4.5 to 6.5 by calcium hydroxide to obtain a first-stage strain, and performing primary screening on the first-stage strain to obtain a primary strain;
s3, primary strain culture: placing the primary strain into a jar filled with the culture solution, performing static culture for 12-24 h at 32-40 ℃, measuring the pH value once every 4h, adjusting the pH value to 4.5-6.5 by calcium hydroxide to obtain a secondary strain, and re-screening the secondary strain to obtain a secondary strain;
s4, secondary strain culture: and (3) placing the secondary strain into a tank filled with the culture solution, carrying out static culture for 24-48 h at the temperature of 32-40 ℃, measuring the pH value once every 4h, and adjusting the pH value to 4.5-6.5 by using calcium hydroxide to obtain the lactic acid bacteria agent.
In S2, primary screening is carried out on the primary strain, and the primary strain is screened out to be not less than 3m2The bacterial colony of (2) is used as a primary strain; in S3, the primary strain is re-screened to screen out not less than 4m2The colonies of (4) were used as secondary species.
As a further optimization of the preparation method of the lactic acid bacteria agent for the industrial non-hydrochloric acid pickled Chinese cabbage, the method also comprises the following steps:
s5, strain preservation: sterilizing glycerol under high pressure, mixing sterilized glycerol and third-stage strain at ratio of 1:1, and storing in refrigerator.
As a further optimization of the preparation method of the industrial lactic acid bacteria agent for the pickled Chinese cabbage without the salt, the final concentration of the glycerol is 10-30%.
Has the advantages that: the invention provides a lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage and a preparation method of a lactic acid bacteria agent, wherein the preparation raw material cost of the culture solution is low, the culture solution is easy to obtain, and the fermentation period of the salt-free pickled Chinese cabbage can be shortened; compared with a direct-vat-set lactic acid bacteria agent, the preparation process of the lactic acid bacteria agent is simpler and more convenient, the cost is lower, and the control is convenient.
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FIG. 1 is a flow chart of a method for preparing a lactic acid bacterium agent.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
A lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage comprises the following components in parts by weight: 2-3 parts of sodium acetate, 10-20 parts of glucose or maltose, 0.15-3 parts of aspartic acid, 5-10 parts of beef extract, 1.5-2 parts of calcium carbonate, 15-20 parts of calcium hydroxide, 100-200 parts of tomato juice, 3-5 parts of inorganic salt, 0.5-1 part of tryptone and 2-5 parts of yeast extract.
The lactic acid bacteria culture solution also comprises 20-25 parts of beer liquid; as the inorganic salt, sodium salt and the like can be used.
Among the ingredients, sodium acetate inhibits part of the bacterial growth; glucose or maltose provide a carbon source; the beef extract provides a carbon source, vitamins, nucleotides and trace elements; calcium carbonate is subjected to performance identification on the amount of generated lactic acid, the generated acid is neutralized, and the lactic acid can dissolve the calcium carbonate to form a transparent calcium ring; adjusting the pH value by calcium hydroxide; the tomato juice provides vitamins and alkaline elements and inhibits the growth of bacteria; inorganic salts are used to provide trace elements; tryptone provides a nitrogen source and vitamins; the yeast extract provides nitrogen sources, vitamins, growth factors, and the like.
Example 1
The strain in the embodiment adopts leuconostoc mesenteroides, and the culture solution of the leuconostoc mesenteroides comprises the following components in parts by mass: 3 parts of sodium acetate, 10 parts of glucose or maltose, 0.2 part of aspartic acid, 10 parts of beef extract, 16 parts of calcium carbonate, 15 parts of calcium hydroxide, 150 parts of tomato juice, 3 parts of inorganic salt, 1 part of tryptone and 3 parts of yeast extract.
Example 2
The strain in the embodiment adopts pediococcus pentosaceus, and the culture solution of the pediococcus pentosaceus comprises the following components in parts by mass: 2 parts of sodium acetate, 10 parts of glucose or maltose, 0.15 part of aspartic acid, 8 parts of beef extract, 20 parts of calcium carbonate, 15 parts of calcium hydroxide, 200 parts of tomato juice, 5 parts of inorganic salt, 1 part of tryptone, 2 parts of yeast extract and 20 parts of beer liquor.
Example 3
The strain in the embodiment adopts streptococcus lactis, and the culture solution of the streptococcus lactis comprises the following components in parts by weight: 2.5 parts of sodium acetate, 10 parts of glucose or maltose, 0.3 part of aspartic acid, 10 parts of beef extract, 15 parts of calcium carbonate, 15 parts of calcium hydroxide, 200 parts of tomato juice, 3 parts of inorganic salt, 0.8 part of tryptone, 5 parts of yeast extract and 25 parts of beer liquor.
Example 4
The strain in the embodiment adopts lactococcus lactis subsp lactis, and the culture solution of the lactococcus lactis subsp lactis comprises the following components in parts by weight: 2 parts of sodium acetate, 10 parts of glucose or maltose, 0.15 part of aspartic acid, 10 parts of beef extract, 20 parts of calcium carbonate, 20 parts of calcium hydroxide, 100 parts of tomato juice, 3 parts of inorganic salt, 1 part of tryptone, 3 parts of yeast extract and 20 parts of beer liquor.
The preparation method of the industrial lactic acid bacteria agent for the pickled Chinese cabbage without the salt is suitable for the four embodiments and comprises the following steps:
s1, strain activation: and inoculating the four preserved strains into an MRS culture medium, and carrying out anaerobic culture at 37 ℃ for 24h to obtain activated strains.
According to the difference of four strains, preparing corresponding culture solution and respectively putting the culture solution into four bottles, four jars and four tanks, wherein in the culture process, the temperature condition of leuconostoc mesenteroides is 40 ℃, the temperature condition of pediococcus pentosaceus is 35 ℃, the temperature condition of streptococcus lactis is 40 ℃, and the temperature condition of lactococcus lactis subsp lactis is 37 ℃.
S2, activated strain culture: putting the four activated strains into a bottle filled with corresponding culture solution according to the inoculation amount of 3%, carrying out shake cultivation for 12-24 h under the corresponding temperature condition, measuring the pH value once every 4h, adjusting the pH value to 4.5-6.5 by using calcium hydroxide to obtain four primary strains, and carrying out primary screening on the four primary strains to obtain four primary strains.
S3, primary strain culture: putting the four primary strains into a jar filled with corresponding culture solution according to the inoculation amount of 3%, standing and culturing for 12-24 h under the corresponding temperature condition, measuring the pH value once every 4 hours, adjusting the pH value to 4.5-6.5 by calcium hydroxide to obtain four secondary strains, and re-screening the four secondary strains to obtain four secondary strains.
S4, secondary strain culture: putting the four secondary strains into a tank filled with corresponding culture solution according to the inoculation amount of 3%, carrying out standing culture for 24-48 h under the corresponding temperature condition, measuring the pH value once every 4h, and adjusting the pH value to 4.5-6.5 by using calcium hydroxide to obtain four lactic acid bacteria.
The pH value can be detected by adopting 0.5 percent bromcresol purple, and whether calcium hydroxide needs to be added to adjust the pH value is selected according to the detected result.
S2, primary screening the primary strain, comparing the morphology of the bacterial colony, and screening out strains with a size not less than 3m according to the growth rate of the bacterial colony2The bacterial colony of (2) is used as a primary strain; in S3, the primary strain is re-screened to screen out not less than 4m2Colonies with larger calcium rings were used as secondary species.
The preparation method of the lactic acid bacteria agent also comprises the following steps:
s5, strain preservation: sterilizing glycerol under high pressure, mixing sterilized glycerol and third-stage strain at ratio of 1:1, and storing in refrigerator.
In S5, the final concentration of glycerol is 10-30%, and 25% is adopted here.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A lactic acid bacteria culture solution for industrial salt-free pickled Chinese cabbage is characterized in that: the composite material comprises the following components in parts by mass: 2-3 parts of sodium acetate, 10-20 parts of glucose or maltose, 0.15-3 parts of aspartic acid, 5-10 parts of beef extract, 1.5-2 parts of calcium carbonate, 15-20 parts of calcium hydroxide, 100-200 parts of tomato juice, 3-5 parts of inorganic salt, 0.5-1 part of tryptone and 2-5 parts of yeast extract.
2. The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage as claimed in claim 1, wherein: also comprises 20-25 parts of beer liquid.
3. The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage as claimed in claim 1, wherein: the composite material comprises the following components in parts by mass: 3 parts of sodium acetate, 10 parts of glucose or maltose, 0.2 part of aspartic acid, 10 parts of beef extract, 16 parts of calcium carbonate, 15 parts of calcium hydroxide, 150 parts of tomato juice, 3 parts of inorganic salt, 1 part of tryptone and 3 parts of yeast extract.
4. The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage as claimed in claim 2, wherein: the composite material comprises the following components in parts by mass: 2 parts of sodium acetate, 10 parts of glucose or maltose, 0.15 part of aspartic acid, 8 parts of beef extract, 20 parts of calcium carbonate, 15 parts of calcium hydroxide, 200 parts of tomato juice, 5 parts of inorganic salt, 1 part of tryptone, 2 parts of yeast extract and 20 parts of beer liquor.
5. The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage as claimed in claim 2, wherein: the composite material comprises the following components in parts by mass: 2.5 parts of sodium acetate, 10 parts of glucose or maltose, 0.3 part of aspartic acid, 10 parts of beef extract, 15 parts of calcium carbonate, 15 parts of calcium hydroxide, 200 parts of tomato juice, 3 parts of inorganic salt, 0.8 part of tryptone, 5 parts of yeast extract and 25 parts of beer liquor.
6. The lactic acid bacteria culture solution for the industrial salt-free pickled Chinese cabbage as claimed in claim 2, wherein: the composite material comprises the following components in parts by mass: 2 parts of sodium acetate, 10 parts of glucose or maltose, 0.15 part of aspartic acid, 10 parts of beef extract, 20 parts of calcium carbonate, 20 parts of calcium hydroxide, 100 parts of tomato juice, 3 parts of inorganic salt, 1 part of tryptone, 3 parts of yeast extract and 20 parts of beer liquor.
7. A preparation method of a lactic acid bacteria agent for industrial non-salt pickled Chinese cabbage is characterized by comprising the following steps: the method comprises the following steps:
s1, strain activation: inoculating the preserved strain into an MRS culture medium, and carrying out anaerobic culture at 37 ℃ for 24h to obtain an activated strain;
s2, activated strain culture: placing the activated strain into a bottle filled with the culture solution of any one of claims 1 to 6, performing shake culture at 32 to 40 ℃ for 12 to 24 hours, measuring the pH value every 4 hours, adjusting the pH value to 4.5 to 6.5 by calcium hydroxide to obtain a first-stage strain, and performing primary screening on the first-stage strain to obtain a primary strain;
s3, primary strain culture: placing the primary strain into a jar filled with the culture solution, performing static culture for 12-24 h at 32-40 ℃, measuring the pH value once every 4h, adjusting the pH value to 4.5-6.5 by calcium hydroxide to obtain a secondary strain, and re-screening the secondary strain to obtain a secondary strain;
s4, secondary strain culture: and (3) placing the secondary strain into a tank filled with the culture solution, carrying out static culture for 24-48 h at the temperature of 32-40 ℃, measuring the pH value once every 4h, and adjusting the pH value to 4.5-6.5 by using calcium hydroxide to obtain the lactic acid bacteria agent.
8. The method for preparing the lactic acid bacteria agent for the industrial pickled Chinese cabbage according to claim 7, which comprises the following steps: in S2, primary screening is carried out on the primary strain, and the primary strain is screened out to be not less than 3m2The bacterial colony of (2) is used as a primary strain; in S3, the primary strain is re-screened to screen out not less than 4m2The colonies of (4) were used as secondary species.
9. The method for preparing the lactic acid bacteria agent for the industrial pickled Chinese cabbage according to claim 7, which comprises the following steps: further comprising the steps of:
s5, strain preservation: sterilizing glycerol under high pressure, mixing sterilized glycerol and third-stage strain at ratio of 1:1, and storing in refrigerator.
10. The method for preparing the lactic acid bacteria agent for the industrial pickled Chinese cabbage according to claim 8, which comprises the following steps: the final concentration of the glycerol is 10-30%.
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| CN102894320A (en) * | 2012-11-09 | 2013-01-30 | 黑龙江省轻工科学研究院 | Preparation method of compound lactobacillus-fermented sauerkraut |
| CN104232506A (en) * | 2014-07-17 | 2014-12-24 | 东北林业大学 | Lactic acid bacteria complex inoculant for Chinese sauerkraut fermentation |
| CN104498414A (en) * | 2015-01-19 | 2015-04-08 | 四川农业大学 | Lactobacillus propagation solution preparation method and inoculant preparation method in industrial salted vegetable production |
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| CN102894320A (en) * | 2012-11-09 | 2013-01-30 | 黑龙江省轻工科学研究院 | Preparation method of compound lactobacillus-fermented sauerkraut |
| CN104232506A (en) * | 2014-07-17 | 2014-12-24 | 东北林业大学 | Lactic acid bacteria complex inoculant for Chinese sauerkraut fermentation |
| CN104498414A (en) * | 2015-01-19 | 2015-04-08 | 四川农业大学 | Lactobacillus propagation solution preparation method and inoculant preparation method in industrial salted vegetable production |
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