CN110699273B - Lactobacillus casei and application thereof - Google Patents
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- CN110699273B CN110699273B CN201910689939.6A CN201910689939A CN110699273B CN 110699273 B CN110699273 B CN 110699273B CN 201910689939 A CN201910689939 A CN 201910689939A CN 110699273 B CN110699273 B CN 110699273B
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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
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L23/00—Soups; Sauces; Preparation or treatment thereof
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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
- C12N1/36—Adaptation or attenuation of cells
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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
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/245—Lactobacillus casei
Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus casei and application thereof. The invention discloses lactobacillus casei LHZ-ST-002, which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms with the preservation number of CGMCC NO. 16447; has the characteristics of good acid resistance, strong acid production and strong salt resistance. The invention also provides a preparation method of the lactobacillus casei LHZ-ST-002, which comprises the processes of strain activation, ultraviolet mutagenesis and strain screening. The invention also discloses application of the lactobacillus casei LHZ-ST-002 in preparation of high-salt fermented food, namely sour soup, wherein the lactobacillus casei LHZ-ST-002 is used for preparing the sour soup, and the acidity of the obtained sour soup is 1.2-1.9 times that of the common lactobacillus casei fermented sour soup in the market.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus casei and application thereof.
Background
Lactobacillus casei (l.casei) belongs to the bacterial domain (Bacilli), Firmicutes, Bacilli (Bacilli), Lactobacillales (Lactobacillales), Lactobacillaceae (Lactobacillaceae), is a short-rod-shaped or long-rod-shaped pleomorphic bacillus, and has different lengths, and the common width is less than 1.5 μm. The two ends of the lactobacillus casei are parallel and level and are square, and the arrangement mode is mostly short chain or long chain. Coccoid bacteria are also sometimes seen; gram staining is positive, no motility and no spore generation.
Lactobacillus casei is combined with Lactobacillus acidophilus and Bifidobacterium and is called "healthy three beneficial bacteria". The lactobacillus casei can utilize glucose to generate lactic acid and some functional substances through a glycolysis process, and is widely applied to the fields of food fermentation and preservation, dairy products, medical care and the like; however, lactobacillus casei has poor adaptability and low survival rate in a high-salt environment, so the lactobacillus casei is rarely applied to the fields of high-salt foods such as sour soup, chopped hot peppers, pickled vegetables and the like.
The Guizhou Miao Dong minority sour soup is prepared by taking bean clear liquid or rice soup, tomatoes, hot peppers and the like as raw materials, adding salt, water and seasonings, and fermenting the raw materials by utilizing microorganisms attached to the raw materials or adding common lactobacillus casei strains in an anaerobic environment; is widely favored by consumers due to the rich nutrition and the property of promoting digestion. However, the naturally occurring sour soup has problems in that safety and quality are not stable. The common lactobacillus casei strain has the problems of poor acid resistance and salt resistance, the survival rate in sour soup is low, and the acidity of the prepared sour soup is low. Therefore, people use the common lactobacillus casei and other salt-tolerant and acid-resistant strains for preparing the sour soup by fermentation in a compounding way, and the other salt-tolerant and acid-resistant strains and the common lactobacillus casei can influence each other in the fermentation process, so that the fermentation process of the sour soup is not easy to control, and the quality of the obtained sour soup is not stable; and the cost is high. For example, the patent application No. CN201910160736.8 discloses a method for preparing pure fermented bean acid soup, which is characterized in that fermented bean curd is prepared by mixing lactobacillus rhamnosus, lactobacillus Maydis liquid, lactobacillus paracasei and Kluyveromyces lactisSoaking to obtain sour soup; the acidity of the obtained sour soup is about 10-13 g/L and is still low. For example, three strains with strong acid-producing capability screened from sour soup are researched and used for fermentation of sour soup in the document "development of Miao national white sour soup beverage" (university of Sichuan agriculture, 2014); the screened strain is identified as lactobacillus casei, and the optimal inoculation amount of lactobacillus casei LA for fermentation of white sour soup is 107cfu/ml, and the total acid content can reach 3.03 per mill after sealed fermentation for 7 days at 35 ℃. The lactobacillus casei LA screened in the document has good acid production capability when used for fermentation of white sour soup, but the acidity of the obtained white sour soup is still low, and the flavor is influenced by the fermentation time.
Disclosure of Invention
In order to solve the technical problems in the prior art, the invention provides lactobacillus casei LHZ-ST-002 produced by mutagenesis, and the lactobacillus casei LHZ-ST-002 is applied to high-salt fermented foods such as sour soup, white sour soup and the like.
The invention aims to provide lactobacillus casei LHZ-ST-002 which is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation number is CGMCC NO.16447, the preservation date is 2018, 9 and 10 days, the preservation address is No.1 Hospital of Xilu of Chaoyang district, Beijing, the preservation number is CGMCC NO.16447, and the lactobacillus casei is classified and named.
Preferably, said lactobacillus casei LHZ-ST-002 has the following properties:
has good acid resistance, and the lactobacillus casei LHZ-ST-002 is added according to the proportion of 106Inoculating cfu/L into bean whey fermentation liquid with acidity of 20g/L, and culturing for 26h while maintaining viable count at 106-108.15Between cfu/L, as shown in FIG. 4;
has strong acid-producing and salt-tolerant abilities, and the lactobacillus casei LHZ-ST-002 is added according to the proportion of 106cfu/L inoculated in bean clear liquid with salt concentration of 4-8wt% and cultured for 48 days, the viable count is kept at 104.4-106.05Between cfu/L; the Lactobacillus casei LHZ-ST-002 is added according to the proportion of 106cfu/L is inoculated into bean clear liquid with the salt concentration of 0-4wt% and the acid yield can reach 8.45-8.90g/L after the bean clear liquid is cultured for 48 hours; as shown in fig. 5.
The invention provides a preparation method of lactobacillus casei LHZ-ST-002, which comprises the following steps:
(1) activating strains: inoculating the bean acid broth into MRS liquid culture medium, culturing at 37 deg.C for 2-3 times, and storing at 4 deg.C;
(2) ultraviolet mutagenesis: taking thalli in logarithmic phase from the bacterial liquid stored in the step (1), centrifuging at 7000rpm/min for 5min, collecting thalli, and washing with sterile physiological saline with mass fraction of 0.85% for 2 times; then suspending in physiological saline to obtain 108Preheating the CFU/m L thallus suspension for 30min by turning on a 20W ultraviolet lamp, placing the 5m L thallus suspension in a culture dish, and irradiating for 30s at a distance of 30cm by using the 20W ultraviolet lamp; finally diluting the irradiated thallus suspension to appropriate gradient, spreading 200 μ L on MRS plate, quickly wrapping with kraft paper, and placing in 37 deg.C anaerobic incubator (CO)2:10％、N290 percent) for 48 hours to obtain mutagenic bacteria liquid;
(3) strain screening: centrifuging and washing the mutagenized bacteria liquid at 7000rpm/min for 5min for 2 times, taking thalli precipitates, diluting to a proper gradient, coating 200 mu L of each gradient on an MRS plate added with calcium carbonate and 5% NaCl, placing the MRS plate in an anaerobic incubator at 37 ℃ for culturing for 48h, picking out a bacterial colony with a large calcium-dissolving ring, and placing the bacterial colony in an MRS liquid culture medium for 12h to obtain the LHZ-ST-002 bacterial strain.
Another purpose of the invention is to provide an application of the lactobacillus casei LHZ-ST-002 in preparing high-salt fermented food.
Preferably, the high-salt fermented food product includes, but is not limited to, soups.
Preferably, the lactobacillus casei LHZ-ST-002 is used for fermentation of soups.
The invention also provides a method for preparing the red sour soup by the lactobacillus casei LHZ-ST-002, which comprises the following steps: firstly, inoculating lactobacillus casei LHZ-ST-002 into sterilized fresh soybean clear liquid, and fermenting for 20-24h at 32-36 ℃ to obtain lactobacillus casei LHZ-ST-002 fermentation liquid; then processing and mixing the raw materials for preparing the sour soup and the seasonings to prepare a fermentation stock solution; finally, lactobacillus casei LHZ-ST-002 fermentation liquor is inoculated into the fermentation stock solution and fermented for 10 to 15 days at the temperature of 32 to 36 ℃, and the red sour soup with the acidity of 13.95g/L and above can be obtained.
A method for preparing white sour soup from Lactobacillus casei LHZ-ST-002 comprises the following steps: firstly, inoculating lactobacillus casei LHZ-ST-002 into sterilized fresh soybean clear liquid, and fermenting for 20-24h at 32-36 ℃ to obtain lactobacillus casei LHZ-ST-002 fermentation liquid; then gelatinizing and liquefying the rice flour to obtain rice soup, and spreading for cooling for later use; finally inoculating lactobacillus casei LHZ-ST-002 fermentation liquor into rice soup, and fermenting at 32-36 deg.C for 10-15 days to obtain white sour soup with acidity of 9.42-10.01 g/L.
Preferably, the viable count of the lactobacillus casei LHZ-ST-002 fermentation liquor is 9.1 × 108cfu/mL。
Preferably, the inoculation amount of the lactobacillus casei LHZ-ST-002 fermentation liquor is 3 percent of the volume of the fermentation stock liquor or rice soup.
The invention has the beneficial effects that: the lactobacillus casei LHZ-ST-002 is obtained by extracting from bean acid soup with high acidity (the acidity is more than or equal to 20g/L) and performing ultraviolet mutagenesis and salt tolerance screening (the addition amount of NaCl is 5 percent), and has the characteristics of good acid resistance, and strong acid production and salt tolerance; when the lactobacillus casei LHZ-ST-002 is used for preparing high-salt fermented food, a higher viable count can be maintained in the fermentation process, the viable count of the prepared product is high, and the nutrition is healthier; the acidity of the sour soup prepared by fermenting the lactobacillus casei LHZ-ST-002 is 1.20 to 1.90 times of that of the common lactobacillus casei fermented sour soup; the lactobacillus casei LHZ-ST-002 is obtained by ultraviolet mutagenesis and screening of the traditional bean sour soup in China, and has simple process and low cost.
Drawings
FIG. 1 is a photograph showing the colonies of the present invention, Lactobacillus casei LHZ-ST-002.
FIG. 2 is a photograph of the present invention showing the microscopic examination of the bacterium casei LHZ-ST-002.
FIG. 3 is a phylogenetic tree of Lactobacillus casei LHZ-ST-002 constructed using MEGA5.0 software according to the present invention.
FIG. 4 is a graph showing the growth of the strain of Lactobacillus casei LHZ-ST-002 of the present invention in a whey fermentation broth having an acidity of 20 g/L.
FIG. 5 shows the strain LHZ-ST-002 of the present invention,The growth curve diagram and the bar chart of the acid production condition of the common lactobacillus casei in bean serum with different salt contents; wherein
The acid yield of the lactobacillus casei LHZ-ST-002,
is the acid yield of the common lactobacillus casei,
the number of the viable bacteria of the lactobacillus casei LHZ-ST-002;
is the viable count of common lactobacillus casei.
Detailed Description
The technical solution of the present invention is further limited by the following specific embodiments, but the scope of the claims is not limited to the description.
EXAMPLE 1 preparation and identification of the bacterial species
Preparing strains:
(1) strain activation
Inoculating bean acid broth liquid stored in a laboratory into an MRS liquid culture medium, culturing at 37 ℃, after passage for 2-3 times, and storing at 4 ℃ for later use;
(2) ultraviolet mutagenesis
The thalli in the logarithmic growth phase is taken from the bacteria liquid preserved in the step (1), the thalli is collected by centrifugation at 7000rpm/min for 5min, washed 2 times by using sterile physiological saline with the mass fraction of 0.85% (w/w), and then suspended in the physiological saline to prepare 108CFU/m L cell suspension. Before irradiation, the ultraviolet lamp is turned on to preheat for 30 min. An equal amount of 5m L of the bacterial suspensions are respectively placed in 7 culture dishes and irradiated for 30s under an ultraviolet lamp with the power of 20W and the distance of 30cm (the culture dish cover is opened to start timing, and the culture dish cover is closed immediately after timing is finished). Taking strains irradiated by ultraviolet rays and strains not irradiated by ultraviolet rays, and diluting to 10 degrees-6Taking 200 mul to coat on MRSQuickly wrapping with kraft paper on a flat plate, and culturing in 37 deg.C anaerobic incubator (CO2: 10%, N2: 90%) for 48h to obtain mutagenic bacteria liquid.
(3) Strain screening
Centrifuging and washing the mutagenic bacteria liquid twice at 7000rpm/min for 5min, taking thalli precipitate, and respectively diluting to 10-6And (2) coating 200 mu L of each gradient in an MRS plate added with calcium carbonate and 5% NaCl, setting a control group, culturing for 48h in an anaerobic incubator at 37 ℃, observing and selecting a bacterial colony with a large calcium-dissolving ring, and culturing for 12h in an MRS liquid culture medium to obtain the LHZ-ST-002 bacterial strain.
Preparing the MRS liquid culture medium: 10g of peptone, 10g of beef powder, 5g of yeast extract, 20g of glucose, 5g of sodium acetate, 2g of diammonium hydrogen citrate, 2g of dipotassium hydrogen phosphate, 0.2g of magnesium sulfate, 0.04g of manganese sulfate, 801g of tween and distilled water, adjusting the pH value to 6.2-6.4, and sterilizing at the high temperature of 115 ℃ for 15min for later use.
And (3) identification of strains: morphological and molecular biological identification are carried out on the obtained LHZ-ST-002 strain:
1. identification index
Morphological identification: gram stain, cell morphology, presence or absence of spores
Molecular biological identification 16SrRNA sequence analysis
2. Identification results
Morphological characteristics of LHZ-ST-002 Strain: the colony morphology is a round yellow colony observed by colony and thallus morphology, the surface is smooth and flat, transparent, the edge is neat, and has an obvious calcium dissolving ring, and the thallus morphology is gram positive bacilli, does not move, does not form spores, and is in a net arrangement.
16S rRNA gene sequencing identification: the 16S rRNA gene sequence for detecting the LHZ-ST-002 strain is as follows: CACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAGGTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGT
According to the detected 16S rRNA gene sequence, the sequence length of the strain LHZ-ST-002 is 1377 bp; then, the 6S rRNA gene sequence of LHZ-ST-002 strain was imported into NCBI for sequence alignment and homology analysis, and the results are shown in Table 1:
TABLE 1 alignment of the sequences of Strain KJY11 on CNBI
As can be seen from Table 1, the homology of the nucleotide sequence of LHZ-ST-002 strain with that of Lactobacillus paracasei strain CG2(KU315090.1) is 100%; according to the comparison results in Table 1, a phylogenetic tree was constructed by using MEGA5.0 software, the structure is shown in FIG. 3, and it can be seen from the table that LHZ-ST-002 strain is on one branch with Lactobacillus paracasei strain CG2(KU315090.1), and thus LHZ-ST-002 strain was identified as Lactobacillus casei (Lactobacillus paracasei strain 2-1).
EXAMPLE 2 Studies of the Properties of Lactobacillus casei LHZ-ST-002 acid resistance
The research method comprises the following steps: mixing Lactobacillus casei LHZ-ST-002 according to 106Inoculating cfu/L into bean whey fermentation liquid with acidity of 20g/L, culturing for 26h, detecting viable count of Lactobacillus casei LHZ-ST-002 in bean whey fermentation liquid every 2h, and drawing line drawing with detection result as shown in figure 4.
And (4) analyzing results: as can be seen from the attached FIG. 4, the number of viable bacteria of Lactobacillus casei 2-1 in the whey fermentation broth increased with the increase of the culture time in the first 16h, and reached a maximum of 10 in the 16 th h8.15cfu/L; in the period of 16-26h, the drunk time is increased, the number of the viable bacteria of lactobacillus casei LHZ-ST-002 is reduced, and the change is not obvious; the number of viable bacteria was kept at 10 throughout the culture period6-108.15cfu/L; therefore, the lactobacillus casei LHZ-ST-002 has good acid resistance.
1. Study on acid production and salt tolerance
The research method comprises the following steps: respectively inoculating the same amount of lactobacillus casei LHZ-ST-002 and lactobacillus casei chebyssus into sterilized bean clear liquid with the same volume and the NaCl addition amount of 0, 1 wt%, 2 wt%, 3 wt%, 4wt%, 5 wt%, 6 wt%, 7 wt%, 8wt%, 9 wt% and 10 wt%, wherein the inoculation amount is 106cfu/L, culturing for 48h under the same environment, detecting the acid production and viable count of Lactobacillus casei LHZ-ST-002 and Lactobacillus casei in the soybean clear liquid, and plotting the detection result into a chart, wherein the result is shown in figure 5.
And (4) analyzing results: as can be seen from the attached figure 5, the acid production amount and the viable count of the lactobacillus casei are obviously reduced along with the increase of the NaCl content in the soybean serum, and when the NaCl content is increased to 8wt%, the lactobacillus casei basically does not produce acid, and the viable count and the acid production amount are basically changed to 0. When NaCl content in the soybean clear liquid is increased from 0 to 4wt% L, acid yield and viable count of lactobacillus casei LHZ-ST-002 are changedThe number of viable bacteria is kept at 106-106.1Between cfu/L, is 10 of the viable count of the common lactobacillus casei0.2-102The acid yield is kept between 8.45 and 8.9g/L, which is about 2 to 5 times of that of the common lactobacillus casei; then, with the continuous increase of NaCl content, the acid production amount and the viable count of the lactobacillus casei LHZ-ST-002 tend to be reduced, but when the NaCl content is increased to 8wt%, the lactobacillus casei LHZ-ST-002 also has certain acid production amount and viable count, and the viable count is 104.4cfu/L, the acid yield is 4.12 g/L. In the soybean clear liquid with the same NaCl content, the acid production amount and the viable count of the lactobacillus casei LHZ-ST-002 are obviously higher than those of the common lactobacillus casei. Therefore, the lactobacillus casei LHZ-ST-002 provided by the invention has acid production and salt tolerance obviously superior to those of the common lactobacillus casei, has stronger acid production and salt tolerance, and can be used for fermenting high-salt food.
Example 3 application of Lactobacillus casei LHZ-ST-002 in tomato Red sour soup
Inoculating the obtained lactobacillus casei LHZ-ST-002 into sterilized fresh soybean clear liquid, and fermenting at 32-36 deg.C for 20-24h to obtain lactobacillus casei LHZ-ST-002 fermentation liquid with viable count of 9.1 × 108cfu/m. Cleaning fresh tomatoes, and cutting into tomato blocks for later use; grinding the tomatoes into paste by using a colloid mill for later use; inoculating, namely adding 3 percent of prepared lactobacillus casei LHZ-ST-002 fermentation liquor into crushed tomatoes and uniformly mixing; fermenting, namely fermenting the mixed materials at the temperature of 32-36 ℃ for 10-15 days to obtain the tomato sour soup with the acidity of 13.95-15.76g/L, which is 1.25-1.54 times higher than that of the common lactobacillus casei fermented tomato sour soup in the market (6.20 g/L).
Example 4 application of Lactobacillus casei LHZ-ST-002 in Hot Pepper Red sour soup
Inoculating the obtained lactobacillus casei LHZ-ST-002 into sterilized fresh soybean clear liquid, and fermenting at 32-36 deg.C for 20-24h to obtain lactobacillus casei LHZ-ST-002 fermentation liquid with viable count of 9.1 × 108cfu/m. Cleaning fresh pepper and chopping for later use; preparing salt, ginger, garlic and rice wine according to related proportions for later use; will be at the topPulverizing the Capsici fructus, rhizoma Zingiberis recens, and Bulbus Allii into paste with colloid mill; mixing, namely uniformly mixing the prepared hot pepper, ginger, garlic, salt and rice wine; inoculating, namely adding 3 percent of prepared lactobacillus casei LHZ-ST-002 fermentation liquor into the mixed liquor; fermenting, namely fermenting the mixed materials at the temperature of 32-36 ℃ for 15 days to obtain the red pepper sour soup with the acidity of 24.30-26.12g/L, which is 1.7-1.90 times higher than that of the common lactobacillus casei fermented red pepper sour soup in the market (9.00 g/L).
Example 5 application of Lactobacillus casei LHZ-ST-002 in white sour soup
Inoculating the lactobacillus casei LHZ-ST-002 obtained by the invention into sterilized fresh soybean clear liquid, and fermenting for 20-24h at the temperature of 32-36 ℃ to obtain lactobacillus casei LHZ-ST-002 fermentation liquor, wherein the viable count is 9.1 multiplied by 108cfu/ml. Pulverizing fresh rice at high speed to obtain rice flour; gelatinizing and liquefying rice flour to obtain rice soup, and spreading for cooling; the mass concentration of the rice flour in the rice soup is about 1.4%; inoculating, namely adding 3 percent of prepared lactobacillus casei LHZ-ST-002 fermentation liquor into prepared rice soup; fermenting, namely fermenting the mixed materials at the temperature of 32-36 ℃ for 10-15 days to obtain rice sour soup with acidity of 9.24-10.01g/L, which is 1.20-1.38 times higher than that of the rice sour soup (4.20g/L) fermented by lactobacillus casei commonly used in the market.
Comparative example 1
The strain lactobacillus casei LA with strong acid production capacity is screened from the white sour soup according to a strain screening method provided in the development of Miao national white sour soup beverage (Sichuan university of agriculture, 2014). Inoculating the screened lactobacillus casei Del into sterilized fresh bean clear liquid, and fermenting at 32-36 ℃ for 20-24h to obtain lactobacillus casei LA fermentation liquid. Taking the rice soup prepared in the example 5, adding 3% of the prepared lactobacillus casei LA fermentation liquor into the prepared rice soup; fermenting, namely fermenting the mixed materials at the temperature of 32-36 ℃ for 10-15 days until the acidity of the rice acid soup is 5.06-6.12 g/L. The acidity of the fermented rice sour soup is lower than that of the fermented rice sour soup prepared by the lactobacillus casei LHZ-ST-002 in the example 5, and the fermented rice sour soup has obvious difference.
Experimental example 1
Samples of the prepared rice sour soup (white sour soup) of example 5 and comparative example 1 were taken, randomly numbered, and 10 persons trained in relevant food assessment techniques were asked to evaluate the color, taste, texture, etc. of the rice sour soup. The evaluation criteria are shown in Table 2, and the results are shown in Table 3:
TABLE 2 sensory evaluation criteria for white sour soup
TABLE 3 sensory evaluation results
As can be seen from the table, the rice sour soup prepared in comparative example 1 and example 5 has no obvious difference in color and texture; however, the rice sour soup prepared in example 5 is superior to the rice sour soup prepared in comparative example 1 in taste and flavor.
It should be noted that the above examples and test examples are only for further illustration and understanding of the technical solutions of the present invention, and are not to be construed as further limitations of the technical solutions of the present invention, and the invention which does not highlight essential features and significant advances made by those skilled in the art still belongs to the protection scope of the present invention.
Sequence listing
Sequence listing
<110> Guizhou Lianghua Zizhai Biotechnology Co., Ltd
<120> lactobacillus casei and application thereof
<130>
<160>1
<170>PatentIn version 3.5
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<211>1377
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<213>Lactobacillus casei
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CACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAGGTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGT
1377
Claims (9)
1. Lactobacillus casei (L.casei) (L.casei)Lactobacillus casei) LHZ-ST-002 is characterized in that the lactobacillus casei is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation date is 2018, 9 and 10 days, the preservation address is No.1 Siro Sai Lu of the Chaoyang district in Beijing, the preservation number is CGMCC number 16447, and the lactobacillus casei is classified and named.
2. Lactobacillus casei LHZ-ST-002, as claimed in claim 1, wherein the lactobacillus casei LHZ-ST-002 grows well in whey fermentation broth with acidity of 20 g/L.
3. The lactobacillus casei LHZ-ST-002 as claimed in claim 1, wherein the lactobacillus casei LHZ-ST-002 has strong acid and salt tolerance, grows well in bean serum with salt concentration of 4-8wt%, and can generate 8.45-8.90g/L acid after being cultured for 48h in bean serum with salt concentration of 0-4 wt%.
4. Lactobacillus casei LHZ-ST-002, as claimed in claim 1, for use in the preparation of high salt fermented food products.
5. Lactobacillus casei LHZ-ST-002 in accordance with claim 4, wherein the high-salt fermented food product includes but is not limited to soups.
6. A method for the preparation of a red sour soup with lactobacillus casei LHZ-ST-002 as claimed in claim 1, which comprises the following process: firstly, inoculating lactobacillus casei LHZ-ST-002 into sterilized fresh soybean clear liquid, and fermenting for 20-24h at 32-36 ℃ to obtain lactobacillus casei LHZ-ST-002 fermentation liquid; then processing and mixing the raw materials for preparing the sour soup and the seasonings to prepare a fermentation stock solution; finally, the lactobacillus casei LHZ-ST-002 fermentation liquor is inoculated in the fermentation stock solution and fermented for 10 to 15 days at the temperature of 32 to 36 ℃, and the red sour soup with the acidity of more than 13.95g/L can be obtained.
7. A method for preparing white sour soup from lactobacillus casei LHZ-ST-002 as claimed in claim 1, which comprises the following steps: firstly, inoculating lactobacillus casei LHZ-ST-002 into sterilized fresh soybean clear liquid, and fermenting for 20-24h at 32-36 ℃ to obtain lactobacillus casei LHZ-ST-002 fermentation liquid; then gelatinizing and liquefying the rice flour to obtain rice soup, and spreading for cooling for later use; finally inoculating lactobacillus casei LHZ-ST-002 fermentation liquor into rice soup, and fermenting at 32-36 deg.C for 10-15 days to obtain white sour soup with acidity of 9.42-10.01 g/L.
8. Preparation of Lactobacillus casei LHZ-ST-002 according to claim 6Method for preparing sour soup with red color or the method for preparing sour soup with white color from lactobacillus casei LHZ-ST-002 as claimed in claim 7, wherein the viable count of the lactobacillus casei LHZ-ST-002 fermentation liquor is 9.1 x 108 cfu/ml。
9. The method for preparing red sour soup with lactobacillus casei LHZ-ST-002 as claimed in claim 6 or the method for preparing white sour soup with lactobacillus casei LHZ-ST-002 as claimed in claim 7, wherein the inoculation amount of the lactobacillus casei LHZ-ST-002 fermentation liquor is 3% of the volume of the fermentation stock liquor or rice soup.
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