CN116286537A - Lactobacillus paracasei GF045 with auxiliary weight-losing effect and application thereof - Google Patents
Lactobacillus paracasei GF045 with auxiliary weight-losing effect and application thereof Download PDFInfo
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- CN116286537A CN116286537A CN202310335478.9A CN202310335478A CN116286537A CN 116286537 A CN116286537 A CN 116286537A CN 202310335478 A CN202310335478 A CN 202310335478A CN 116286537 A CN116286537 A CN 116286537A
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- lactobacillus paracasei
- sour meat
- weight
- cholesterol
- millet
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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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
Abstract
The invention discloses lactobacillus paracasei GF045 with auxiliary weight-losing effect and application thereof, and the strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.26482. The lactobacillus paracasei GF045 with the auxiliary weight-losing effect has the advantages of good acid resistance, bile salt resistance and safety, good cholesterol and triglyceride degradation capability and obvious weight-losing effect; the lactobacillus paracasei GF045 can be used for preparing the sour meat with moderate acidity, good taste and relatively low fat and cholesterol content, and the preparation method of the sour meat has high production efficiency and is beneficial to industrial production. The lactobacillus paracasei GF045 provided by the invention can be further applied to preparing foods, health-care foods or medicines with the auxiliary weight-losing effect, and can also be used for fermenting sour meat.
Description
Technical Field
The invention belongs to the field of microbial engineering, and relates to probiotics and application thereof, in particular to lactobacillus paracasei GF045 with an auxiliary weight-losing effect and application thereof.
Background
Over the last decade, with the increasing level of living and changing dietary structure of people, the overweight/obese population ratio has increased and gradually progressed to younger. Obesity can cause fat accumulation in vivo, and increase triglyceride and total cholesterol levels, and triglyceride and cholesterol levels are high, which can increase occurrence of obesity related diseases such as hyperlipidemia, coronary heart disease, type 2 diabetes, hypertension, nonalcoholic fatty liver, etc. Obesity and its complications not only pose a serious threat to health, but also pose a heavy burden on the socioeconomic and medical health system, so obesity and related chronic diseases at this stage have become global significant public health problems.
Currently, obesity treatment mainly includes two approaches, lifestyle intervention and medical treatment. Among these, the medical means mainly include the use of drugs, bariatric surgery and intervention of intestinal flora. Lifestyle intervention is the basis of all weight loss plans, but weight loss is limited in magnitude and weight rebound is easy to occur. The medicine has the defects of poor effectiveness, large side effect, long administration period and the like, and can cause weight rebound even if the weight is maintained by restricting diet after stopping administration. Obesity surgery is one of the most effective weight-losing means at present, although the effect of the surgery treatment is remarkable, risks exist, such as general anesthesia risks and surgery risks, and the surgery risks mainly comprise bleeding, digestive tract fistula, infection, internal hernia, postoperative gastroesophageal reflux, nutritional disorder, obesity and other risks.
Numerous studies have found that the intestinal flora is closely related to the onset of obesity. Probiotics can have beneficial effects on human health by mediating changes in the intestinal flora. The regulation of intestinal flora is an effective strategy for improving and managing the obesity, and the therapeutic intervention of probiotics has the characteristics of zero side effect, zero rebound, no water reduction, no skin injury, no diet, no movement, simple and convenient use and the like, and can provide a new treatment method for the obesity.
At present, commercial probiotics strains capable of reducing fat accumulation or reducing weight are almost monopolized by foreign enterprises, such as bifidobacterium animalis B420 of DuPont Dannique, lactobacillus amylovorus CP1563 of Japanese cobicism, bifidobacterium breve B-3 of Japanese forest Yongru industry and the like, and the current research of screening probiotics with the function of losing weight in China is still in an early stage, so that the development of more novel probiotics strains with the function of losing weight and reducing fat has important social significance.
Disclosure of Invention
The invention aims to provide a probiotic with an auxiliary weight-losing effect, namely lactobacillus paracasei GF045, which has good in-vitro tolerance and safety, can remarkably reduce the weight of rats, reduce fat accumulation in vivo, reduce body fat ratio, effectively reduce the index level of total cholesterol, triglyceride and low-density lipoprotein cholesterol in blood fat, and effectively reduce the weight and body fat rate of obese people;
another object of the present invention is to provide an application of lactobacillus paracasei GF045 with auxiliary weight loss effect as described above for preparing sour meat to achieve industrial application of the strain;
the invention also provides a preparation method of the sour meat, which is simple and convenient, has high production efficiency and is beneficial to industrial production;
it is still another object of the present invention to provide another application of the above Lactobacillus paracasei GF045, which can be widely used for preparing foods, health foods or medicines with an auxiliary weight-reducing effect.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
lactobacillus paracasei GF045 with the auxiliary weight-losing effect is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.26482 and the Latin name of Lactobacillus paracasei.
As a limitation to the Lactobacillus paracasei GF045 having the effect of assisting weight loss, the gene sequence of 16SrDNA of this bacterium is as follows:
GAGTTCTCCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAA
CGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGG
GGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGC
ATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCG
CGGCGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGATGATAC
GTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCC
CAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAA
GTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAA
ACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTG
ACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGG
TAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAG
CGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGG
AAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAA
CTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGG
CGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGG
GTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAA
TGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTA
AGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATT
GACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAAC
GCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAG
GTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCT
CGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATG
ACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACA
AACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACC
TGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGC
GAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGC
AACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGC
CGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATG
AGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTTTT。
the invention also provides an application of the lactobacillus paracasei GF045 with the auxiliary weight-losing effect, and the strain can be used for preparing foods, health-care foods or medicines with the auxiliary weight-losing effect.
The invention also provides another application of the lactobacillus paracasei GF045 with the auxiliary weight-losing effect, and the strain can be used for preparing sour meat.
The invention also provides a preparation method of the sour meat, which comprises the following steps: placing pork in a solution A containing glucose and millet juice, sealing, sterilizing, inoculating lactobacillus paracasei GF045, and fermenting until the pH is 4.3-4.5 to obtain sour meat.
As a limitation to the above preparation method, the mass ratio of glucose to millet juice in the solution a is 1: 50-100, wherein the mass ratio of pork to the solution A is 1:2 to 2.5.
As a further limitation to the above preparation method, the preparation method of the millet juice comprises the steps of adding millet into water for boiling, stopping boiling when the specific gravity of the juice is within the range of 1.05-1.35 g/mL at 35-37 ℃, and removing the millet to obtain the millet juice.
As a second limitation of the above preparation method, the sterilization temperature is 75 to 80 ℃ and the time is 3 to 5min.
As a third limitation to the above-mentioned production method, the viable count of the Lactobacillus paracasei GF045 is 4.0X10 9 ～8.0×10 9 cfu/mL, the inoculation amount is 3-10%.
The third limitation of the preparation method is that the fermentation time is 72-120 h, and the fermentation temperature is 35-37 ℃.
By adopting the technical scheme, compared with the prior art, the invention has the following technical progress:
(1) the lactobacillus paracasei GF045 with the auxiliary weight-losing effect has good acid and bile salt resistance, high safety, and meanwhile, has the specific capacity of degrading cholesterol and triglyceride, remarkably reduces the weight and fat/body ratio of an obese rat model, remarkably reduces the TG, TC, LDL-C level of the obese rat model, has obvious weight-losing effect on people and the like;
(2) compared with the traditional medicine for losing weight, the lactobacillus paracasei GF045 provided by the invention has the advantages of no side effect, zero rebound, no water reduction, no harm to skin, no diet, and simpler and more convenient use, and the beneficial lipoprotein-high density lipoprotein content of the organism is higher than that of the traditional weight-losing medicine;
(3) the preparation method of the lactobacillus paracasei GF045 sour meat provided by the invention is determined by combining the characteristics of lactobacillus paracasei GF045 strain. Firstly, the millet paste contains rich high-quality proteins, fat, carbohydrate, vitamins, inorganic salts and the like, and the millet paste juice is adopted to provide propagation nutrients for lactobacillus paracasei GF045 in the preparation method; secondly, determining a fermentation temperature according to the optimal growth temperature of lactobacillus paracasei GF045, and determining a fermentation end point according to a fermentation pH value; the preparation method provided by the invention combines the traditional sour meat fermentation technology by utilizing the modern microorganism fermentation technology, the working efficiency is improved by more than 2 times, the fat and cholesterol content of the prepared sour meat is lower, and the product meets the requirements of modern consumers on healthy diet;
(4) the lactobacillus paracasei GF045 provided by the invention has potential wide application fields, and can be used as a raw material for preparing foods, health-care foods or medicines with the auxiliary weight-losing effect.
In conclusion, the lactobacillus paracasei GF045 provided by the invention has good acid and bile salt resistance and high safety, and also has good cholesterol and triglyceride degradation capability, so that the weight-losing effect is remarkable; the lactobacillus paracasei GF045 can be used for preparing the sour meat with moderate acidity, good taste and relatively low fat and cholesterol content, and the preparation method of the sour meat has high production efficiency and is beneficial to large-scale industrial production.
The lactobacillus paracasei GF045 provided by the invention is a probiotic with a weight-losing function, and can be further applied to preparing foods, health-care foods or medicines with an auxiliary weight-losing function, and can also be used for fermenting sour meat.
Drawings
FIG. 1 is a macro morphology of Lactobacillus paracasei GF045 according to example 1 of the present invention;
FIG. 2 is a microscopic morphology of Lactobacillus paracasei GF045 according to example 1 of the present invention.
Detailed Description
The invention is further illustrated by the following examples. It should be understood that the described embodiments are only for explaining the present invention and do not limit the present invention.
Materials, reagents, and the like used in the examples of the present invention are commercially available unless otherwise specified. The experimental methods for which specific conditions are not specified in the examples are generally conducted under conventional conditions or under conditions recommended by the manufacturer.
EXAMPLE 1 Lactobacillus paracasei GF045
The example includes identification and preservation of lactobacillus paracasei GF045, in vitro tolerability experiments, safety assessment, in vitro cholesterol degradation capability studies, in vitro triglyceride degradation capability studies, and effects on in vivo fat and related indicators.
(1) Identification and preservation of Lactobacillus paracasei GF045 strain
The strain provided by the invention is obtained by separating from yoghurt self-made by a grazing plain herdsman of an inner Mongolian autonomous region by the inventor, and is named as GF045. The GF045 strain is Lactobacillus paracasei (Lactobacillus paracasei) which is identified by China food fermentation industry institute, namely, the inventor stores the strain to China general microbiological culture Collection center with the preservation number of CGMCC No.26482. The biological properties and identification of the GF045 strain are based on the following.
(1) Morphological features
Culturing strain GF045 in MRS culture medium at 37deg.C for 48 hr, wherein the macroscopic morphology of strain GF045 is shown in figure 1, and the colony is white, round, moist, opaque and regular in edge as seen in figure 1; the microscopic morphology of the GF045 strain was cultivated in MRS medium at 37℃for 48 hours, and as shown in FIG. 2, the cells were rod-shaped, 0.5 to 0.7. Mu.m.times.1.0 to 2.2. Mu.m, arranged singly or in pairs, and were gram-positive, as seen from FIG. 2.
(2) Physiological and biochemical characteristics
The physiological and biochemical characteristics of the GF045 strain are shown in table 1.
TABLE 1 VITEKANC identification card for GF045 Strain
The symbols indicate "+", positive; "w", weak positive; "-", negative.
(3) Biological identification
Sequencing the 16SrDNA and pheS genes of the GF045 strain, and respectively comparing the sequences with a professional database taking the NCBI database as a base source to determine that the GF045 strain is lactobacillus paracasei and the Latin name is Lactobacillus paracasei.
Wherein, the 16SrDNA gene sequence of GF045 is as follows:
GAGTTCTCCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTG
ACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGG
TAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAG
CGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGG
AAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAA
CTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGG
CGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGG
GTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAA
TGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTA
AGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATT
GACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAAC
GCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAG
GTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCT
CGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATG
ACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACA
AACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACC
TGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGC
GAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGC
AACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGC
CGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATG
AGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTTTT；
the pheS gene is as follows:
AAGAGACTTTTTATATTACCAATGAGTTACTCATGCGCTCGCAGACAAGTCCAATGCAGGCGCGGACAATGGAAAAGCACGACTTTACCAAAGAACCGCTGAAAATGATTAGCCCTGGGGTGGTTTATCGACGTGATGACGACGATGCTACTCATAGCCATCAGTTTCACCAGATGGAAGGACTCGTCATTGACAAGCATATAACCATGGCTGATCTAAAGGGAACCTTGTTGGCCATGTGCCAACACGTTTTTGGTAAAGATCGGACAATTCGCTTGCGGCCAAGTTATTTTCCATTTACGGAGCCATCCGTTGAAGTTGATGTTTCCTGTTTTCGTTGCGGCGG。
(4) preservation of bacterial species
The inventor stores lactobacillus paracasei GF045 strain to China general microbiological culture Collection center with the preservation number of CGMCCNo.26482 in 2023, 01 and 13.
(2) In vitro tolerance experiment of Lactobacillus paracasei GF045
The part is a lactobacillus paracasei GF045 acid and bile salt resistant characteristic experiment, and an acid resistant experiment and a bile salt resistant experiment of the strain are respectively carried out, and the method comprises the following steps.
(1) Acid resistance test
S1, preparation of artificial gastric juice
Diluting hydrochloric acid with volume fraction of 10% with deionized water, respectively adjusting pH to 2.0, 3.0 and 4.0, adding pepsin to make its final concentration be 1%, and adjusting pH to 2.0, 3.0 and 4.0 with hydrochloric acid. Filtering with microporous membrane of 0.22 μm in a sterile operation table;
s2, activation of Lactobacillus paracasei GF045
S21, taking the Lactobacillus paracasei GF045 cryopreservation tube out of the refrigerator, melting at room temperature, inoculating 100 mu L of the tube into 10mL of sterilized MRS culture medium, and culturing at 37 ℃ for 24 hours to obtain first-generation bacterial liquid;
s22, inoculating the first-generation bacterial liquid into a sterilized MRS culture medium in an inoculum size of 3%, and culturing for 12 hours at 37 ℃ to obtain a second-generation bacterial liquid;
s23, taking 10mL of secondary bacterial liquid, and measuring lactobacillus paracasei in the secondary bacterial liquidThe viable count of GF045 was 1X 10 8 Centrifuging CFU/mL at 8000 r/min for 10min, and separating to obtain thallus;
each activated strain requires microscopic examination to confirm that the strain is pollution-free;
s3, preparation of bacterial suspension
The cells prepared in the step S23 were washed 3 times with 0.85% physiological saline, and a bacterial suspension was prepared.
S4, acid resistance test
Mixing 0.5mL of bacterial suspension with 4.5mL of artificial gastric juice with pH value of 2.0, 3.0 and 4.0 respectively, standing and culturing in a 37 ℃ incubator, sampling after 2h and 4h respectively, diluting and coating a flat plate, culturing for 48h at 37 ℃, and measuring the colony number. The survival rate was calculated using the number of colonies not treated with artificial gastric juice as a control, and the results of the acid-resistance experiment of Lactobacillus paracasei GF045 are shown in Table 2 below.
TABLE 2 Lactobacillus paracasei GF045 acid resistance test results
Note that: colony count without artificial gastric juice treatment was 3.59X10 pairs 7 CFU/mL。
As can be seen from the experimental results in table 2, lactobacillus paracasei GF045 showed very good tolerance in artificial gastric juice, and at pH 2.0, 34.26% of viable bacteria survived after 4 hours of tolerance treatment; when the pH is 3.0, the survival rate of the viable bacteria in the artificial gastric juice reaches 71.59 percent; at a pH of 4.0, lactobacillus paracasei GF045 was hardly lost. Gastric acid can reach 3.5-4.0 after meals, and gastric emptying time is about 4 hours, so that lactobacillus paracasei GF045 is eaten by the human body after meals, and the gastric acid can be effectively assisted to reach intestinal tracts through gastric juice.
(2) Experiment of bile salt resistance
S1, preparation of bile salt solution
Preparing MRS culture medium with concentration of bile salt of 0.1%,0.2% and 0.3% respectively;
s2, activation of Lactobacillus paracasei GF045
Taking out the preserved lactobacillus paracasei GF045 from the glycerol tube for resuscitation, taking 300 mu L of the lactobacillus paracasei GF045, inoculating the 300 mu L of the lactobacillus paracasei to 10mL of MRS liquid culture medium, and culturing the lactobacillus paracasei at 37 ℃ for 16 hours to obtain first-generation bacterial liquid; inoculating the first-generation bacterial liquid into a sterilized MRS culture medium with an inoculum size of 3%, and culturing at 37 ℃ for 16 hours to obtain a second-generation bacterial liquid; each activated strain requires microscopic examination to confirm that the strain is free of contamination.
S3, experiment of bile salt resistance
Inoculating the cultured second-generation bacterial liquid into 50mL culture medium containing bile salts with different concentrations respectively according to the inoculation amount of 2%, and culturing for 2h and 4h at 37 ℃. Sampling, gradient dilution, coating MRS plate, culturing at 37 ℃ for 24 hours, and measuring colony number. The viability was calculated using the number of colonies not treated with the bile salt solution as a control, and the results of the Lactobacillus paracasei GF045 bile salt tolerance test are shown in Table 3 below.
TABLE 3 results of Lactobacillus paracasei GF045 bile salt tolerance experiments
Note that: colony count without bile salt solution treatment was 4.73X10 pairs 7 CFU/mL。
From the experimental results in Table 3, it is found that Lactobacillus paracasei GF045 is hardly lost at a bile salt concentration of 0.1%; at 0.2% bile salt concentration, survival rate exceeded 60%; at 0.3% bile salt concentration, the survival rate decreases rapidly, but the survival rate of the living bacteria exceeds 25%, so that the strain with very good tolerance in probiotics is obtained.
(3) Safety evaluation of Lactobacillus paracasei GF045
(1) Antibiotic susceptibility test
Whether bacteria are sensitive to antibiotics is an important indicator for evaluating bacterial safety, in this example
The 19 antibacterial agent paper sheets are used for detecting the drug sensitivity of lactobacillus paracasei GF045, and the drug sensitivity of the tested strain is judged according to the judgment standard of sensitivity or drug resistance of NCCLS 7 th edition provided by WHO, and the specific measurement results are shown in table 4.
TABLE 4 Lactobacillus paracasei GF045 antibiotic susceptibility test results
Note that: d represents diameter (mm); MGD represents sensitivity; s represents sensitivity; m represents moderate sensitivity; r represents resistance
From the experimental results in table 4 above, it can be seen that: lactobacillus paracasei GF045 showed sensitivity to most antibiotics, only moderately sensitive to trimethoprim (SXT), only resistant to Ciprofloxacin (CIP), indicating a high safety of lactobacillus paracasei GF045.
(2) Hemolysis experiment
Lactobacillus paracasei GF045 was inoculated with an inoculating loop in sheep blood plates under sterile conditions and incubated at 37℃for 48h to observe haemolysis. Hemolysis can form three features on the blood platelets:
alpha hemolysis: grass green hemolytic rings appear around colonies, which are generally opportunistic;
beta hemolysis: a wide transparent hemolytic ring appears around the colony, and the colony is generally strong in pathogenicity;
gamma hemolysis: there were no hemolytic rings around the colonies, and in general the strains were nonpathogenic.
In this example, staphylococcus aureus (Staphylococcus aureus) ATCC25923 was used as a positive control, and after culture, no hemolytic ring appeared around each colony of lactobacillus paracasei GF045, so that lactobacillus paracasei GF045 was gamma-hemolyzed and had no hemolytic activity.
(4) Investigation of ability of Lactobacillus paracasei GF045 to degrade cholesterol in vitro
S1 preparation of cholesterol-containing MRS liquid culture Medium (MRS-CHO)
Preparing a cholesterol solution with concentration of 10mg/mL by using absolute ethyl alcohol, filtering and sterilizing by using a microporous filter membrane with pore diameter of 0.45 mu m, and adding the solution into a sterilized MRS liquid culture medium containing 0.3% of bovine bile salt to make the final concentration of cholesterol be 100 mu g/mL;
s2, determination of cholesterol degradation capacity of lactobacillus paracasei GF045
The viable count of the live bacteria prepared in the step S23 under the item of (1) the acid resistance test in the in vitro tolerance test of (2) Lactobacillus paracasei GF045 in this example was 1X 10 8 Inoculating CFU/mL second-generation bacterial liquid into MRS-CHO liquid culture medium according to an inoculum size of 3%, culturing at 37 ℃ for 12, 24, 36 and 48 hours, and centrifuging the culture liquid for 10min under 4000r/min to obtain supernatant 1;
adding absolute ethyl alcohol into the supernatant 1, mixing uniformly, standing for 5min, centrifuging for 10min at 4000r/min to obtain supernatant 2, measuring absorbance of the supernatant 2 at 510nm by using a cholesterol measurement kit (purchased from Shanghai enzyme-linked biotechnology Co., ltd.), and taking a non-inoculated cholesterol medium as a blank control;
the cholesterol degradation rate was calculated according to the method described in the kit instructions, wherein the formula of the cholesterol degradation rate is shown in formula 1.
Cholesterol degradation rate (%) = (a-B)/a×100% formula 1
In equation 1: a is the absorbance of supernatant fluid when no bacteria are inoculated; b is the absorbance of the supernatant after inoculation.
The cholesterol degradation rates of Lactobacillus paracasei GF045 at various measurement times were calculated as shown in Table 5 below.
TABLE 5 results of in vitro cholesterol degradation rates of Lactobacillus paracasei GF045
| Measurement time (h) | Cholesterol degradation rate (%) |
| 12 | 15.55±1.11 |
| 24 | 20.20±1.68 |
| 36 | 31.45±2.09 |
| 48 | 33.18±3.10 |
As can be seen from the data in table 5: in a cholesterol medium containing 100 mug/mL, lactobacillus paracasei GF045 shows good cholesterol degradation capability, and the cholesterol degradation rate is obviously enhanced within 12-36 hours along with the time extension. At 12h, 15.55% of cholesterol in the medium was degraded; at 36h 31.45% of the cholesterol in the medium was degraded, whereas at 48h lactobacillus paracasei GF045 33.18% of the cholesterol in the medium was degraded, and it was seen that after 36h no significant improvement in degradation capacity occurred, possibly because the shortage of nutrients in the medium was insufficient to support the growth of lactobacillus paracasei.
(5) Investigation of in vitro degradation ability of Lactobacillus paracasei GF045 to triglyceride
S1 preparation of triglyceride-MRS culture medium
Adding 1mL of triglyceride into a 50mL volumetric flask, using absolute ethyl alcohol to fix the volume to prepare triglyceride solution with volume fraction of 2%, ultrasonically dissolving 30min, filtering by using a 0.45 mu m microporous filter membrane, sucking 0.5mL to 9.5mL of liquid MRS culture medium, and sterilizing by high-pressure steam to obtain sterilized triglyceride-MRS culture medium;
s2, determination of degradation capability of lactobacillus paracasei GF045 on triglyceride
Under aseptic conditions, the viable count of the live bacteria prepared in the step S23 under the (1) acid resistance test item in the in vitro tolerance test of lactobacillus paracasei GF045 of (2) this example was 1×10 8 Adding the second-generation bacterial liquid of CFU/mL into a sterilized triglyceride-MRS liquid culture medium according to the inoculation amount of 2%, and culturing for 12 hours, 24 hours, 36 hours and 48 hours at the constant temperature of 37 ℃;
taking 2mL of fermentation liquor in a centrifuge tube, and centrifuging at 6000 r/min for 10min to obtain supernatant A; adding 1mL of the supernatant A into a centrifuge tube, adding 1mL of absolute ethyl alcohol, oscillating for 5mi n, centrifuging 10000 r/min for 10min, obtaining a supernatant B, and measuring the triglyceride degradation rate of the supernatant B according to a method described in a triglyceride detection kit instruction book; the triglyceride detection kit of the embodiment is purchased from Shanghai enzyme-linked biotechnology limited company; the formula of the triglyceride degradation rate is shown in formula 2.
Triglyceride degradation rate (%) = (a-B)/a×100% formula 2
In equation 2: a is the absorbance of a non-inoculated control; b is the absorbance of the sample after inoculation.
Calculated triglyceride degradation rates for Lactobacillus paracasei GF045 at various assay times are shown in Table 6 below.
TABLE 6 Lactobacillus paracasei GF045 in vitro triglyceride degradation results Table
From the results shown in Table 6, lactobacillus paracasei GF045 exhibited a good triglyceride degrading ability in a triglyceride-MRS liquid medium at a content of 0.1%, and the triglyceride degrading ability was gradually increased with the increase of the culture time. At 12h, 19.04% of the triglycerides in the medium were degraded; at 36h 61.82% of the triglycerides in the medium were degraded, whereas at 48h the triglycerides in the medium were not significantly reduced, only 62.75% of the triglycerides were degraded, probably because after 36h lactobacillus paracasei GF045 had no support for the entry of nutrients into the decay and no degradation of the triglycerides could be performed.
(6) Effect of Lactobacillus paracasei GF045 on in vivo fat and related index
Based on the evaluation method described in "Jiu, contributing to control of body fat" in the "method for functional test and evaluation of health food (2022 edition)", the influence of Lactobacillus paracasei GF045 on the indexes such as total cholesterol (TG), triglyceride (TC), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) in rat serum was examined in the animal experiment section.
(1) Animal experiment
90 clean SD male adult rats with body weight range of 200+ -20 g are selected, and after normal feeding and adapting to environment for 5d, the rats are randomly divided into 2 groups according to body weight. The first group was 10 animals, and normal maintenance feed was given as a blank group; the other group is 80, high-calorie feed (basal feed 70%, lard 15% and sucrose 15%) is fed for 14d to be used as a model group, the feed intake and the residual feed are recorded every week, the weight is weighed for 1 time, after continuous feeding for 2 weeks, the fat resistant rats with lower weight gain are eliminated, and the remaining 50 rats with higher weight gain are constructed as a rat fat model; the 50 rat obesity models constructed above were randomly divided into 5 groups, which were model group, positive control group, low dose group, medium dose group, and high dose group, respectively.
After the model 2d is completed, each group is dosed, and the dosed gastric lavage doses of the different groups are shown in table 7, wherein the dosed doses are converted according to the dosed doses of animals and humans, and the dosed doses are 5 times of the recommended dose of human bodies. The administration time of the test substances is 6 weeks, and the test substances are infused 1 time a day; during the experiment, the blank control group is fed with common feed, and the model group, the positive control group, the low-dose group, the medium-dose group and the high-dose group are all fed with high-calorie feed; orlistat used for gastric lavage is purchased from Chongqing Huasen pharmaceutical Co., ltd, and is prepared into a solution with the concentration of 120mg/mL during gastric lavage; the Lactobacillus paracasei GF045 used for stomach irrigation is self-made Lactobacillus paracasei GF045 freeze-dried powder, and the viable count is 5.0X10 11 CFU/g, and the viable count is 1.0X10 during gastric lavage 11 CFU/mL of bacterial suspension. During the experiment, the feed and the residual feed were recorded weekly, and the body weight was weighed 1 time.
Table 7 dosing schedules for rats of different groups during the experimental period
| Group of | Dosing regimen |
| Blank control group | 1mL of physiological saline is infused into stomach for 1 time/d |
| Model group | 1mL of physiological saline is infused into stomach for 1 time/d |
| Positive control group | Gastric orlistat, 60mg/kg BW dose, 1 time/d |
| Low dose group | Lactobacillus paracasei GF045, dose 2.0X10 10 cfu/kg BW,1 time/d |
| Medium dose group | Lactobacillus paracasei GF045, dose 6.0X10 10 cfu/kg BW,1 time/d |
| High dose group | Lactobacillus paracasei GF045, dose 1.0X10 11 cfu/kg BW,1 time/d |
Before the first administration, 0.5ml of the model group, the positive control group, the low dose group, the medium dose group and the high dose group are collected by adopting the orbital veins, and serum is centrifugally taken to measure total cholesterol (TG), triglyceride (TC), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C), and an Elisa kit is adopted for detection.
After the last administration of rats, 16 hours on an empty stomach, the rats were weighed, anesthetized with 1% sodium pentobarbital (0.5 ml/100g BW), dissected to obtain perirenal fat, peritesticular fat and weighed, the fat/body ratio was calculated, and blood was collected and centrifuged to obtain serum TG, TC, LDL-C, HDL-C.
Data were processed using SPSS 22.0, using one-way analysis of variance between groups, P <0.05 indicated significant differences, and P <0.01 indicated significant differences. The statistical table of the body weight, fat and body fat ratio of the rats in different groups in the experimental period is shown in table 8, and the statistical table of the blood lipid indexes of the rats in different groups is shown in table 9.
Table 8 statistical table of body weight, fat and body fat ratios of rats of different groups during the experimental period
Note that: * : the variability was significant (P < 0.05) relative to the placebo group; * *: the difference was very significant (P < 0.01) compared to the blank control group
#: relative to the model group, the variability was significant (P < 0.05); # #. The differences were very pronounced relative to the model group (P < 0.01)
From the data in Table 8, it can be seen that: the rat obesity model is successfully constructed, the weight of the lactobacillus paracasei GF045 low-dose group rats is obviously different from that of the model group rats at the 5 th week, and the difference is extremely obvious at the sixth week; the weights of rats in the medium dose group and the high dose group were significantly different from those of rats in the model group at week 4, and the differences were extremely significant at week 5; the positive control group of rats had a significant difference in weight from the model group at week 3. The three experimental groups and the positive control group have a certain difference in weight loss in the first 3 weeks, but have stronger weight loss effect with the positive control group in the 6 th week, and the high-dose group has similar effect with the positive control group. In addition, the body fat weight and body fat of the three experimental groups were significantly reduced compared to the model group. From this, lactobacillus paracasei GF045 has a better weight-reducing effect on the rat obesity model.
TABLE 9 statistics of blood lipid indicators for rats of different groups
Note that: * : relative to the blank control group, the variability was significant (P < 0.05) #: relative model group, the difference is significant (P < 0.05)
As can be seen from the data in table 9 above, four indices TG, TC, LDL-C, HDL-C of the low model group, the positive control group, the low dose group, the medium dose group and the high dose group showed significant differences (P < 0.05) relative to the blank control group before the gastric drug orlistat or lactobacillus paracasei GF045 was administered after the rat obesity model was established; after the positive control group, the low dose group, the medium dose group and the high dose group are respectively fed with respective samples for 6 weeks, the three indexes of TG, TC, LDL-C are obviously reduced (P < 0.05) relative to the model group, and the indexes of HDL-C are obviously increased (P < 0.05) relative to the model group. Clinically, the high-density lipoprotein level is generally reflected by detecting the content of high-density lipoprotein cholesterol (HDL-C), and the main function of the high-density lipoprotein is to transfer cholesterol in tissues except the liver to the liver for catabolism so as to prevent free cholesterol from accumulating in arterial walls and other tissues, so that the high-density lipoprotein is beneficial to organisms in a normal range; the data in this experiment show that lactobacillus paracasei GF045 can raise the high density lipoprotein level in the blood to some extent, and that the medium and high dose groups have an ability to raise the high density lipoprotein level even more than the well-known OTC slimming drug-orlistat. The experimental result shows that the lactobacillus paracasei GF045 shows the efficacy of assisting in reducing blood fat.
(2) Human body test feeding test
The experiment was carried out according to the evaluation method described under "nine and helps control of body fat" in "method for functional examination and evaluation of health food (2022 edition)".
Subject inclusion criteria: the tested subjects are simple obese people, adult BMI is more than or equal to 30.0, or total fat percentage reaches male >25%, female >30% volunteer subjects.
Subject exclusion criteria: patients with mental diseases complicated with serious diseases such as heart, liver, kidney and hematopoietic system; taking items related to the tested function in a short period of time affects the judgment of the result; the efficacy or safety of the test sample cannot be judged if the test sample is not consumed according to the specification.
In the morning, the body weight and waist circumference of the subject were measured on an empty stomach. Wherein the waistline is the abdomen circumference of the navel. Body fat formula for women: parameter a = waist circumference x 0.74; parameter b = body weight x 0.082+34.8; total body fat weight = a-b; body fat rate= (total body fat weight ≡body weight) ×100%; body fat formula for men: parameter a = waist circumference x 0.74; parameter b = body weight x 0.082+44.7; total body fat weight = a-b; body fat rate= (total body fat weight ≡body weight) ×100%.
The age of the subjects is 20-55 years, 418 people in total. Volunteers were divided into test group 360 and control group 58. Volunteers in the test group consumed the solid beverage 2 bags (2 g each bag containing lactobacillus paracasei GF045 viable count of 2.5x10) after half an hour per day dinner 10 CFU/g), no other type of product was taken during the trial, no product of any type was taken by volunteers in the control group; the preparation method of the solid beverage comprises the following steps: culturing Lactobacillus paracasei GF045 in MRS culture medium, lyophilizing to obtain lyophilized powder, and detecting viable count of 6.2X10 11 cfu/g, uniformly mixing the prepared bacterial powder with sterile fructo-oligosaccharide, inulin and xylo-oligosaccharide according to the mass ratio of 5:15:4:1 to prepare lactobacillus paracasei GF045 with the viable count of 2.5X10 10 The CFU/g solid beverage is packaged into 2 g.
Maintaining regular diet, physical activity and life habit during test food; the test sample was administered for 60 days.
The body weight, waist circumference, body fat rate and other indexes of all volunteers before the test, after the test for 30d and after the test for 60d were accurately recorded, and the specific statistical table is shown in table 10.
Table 10 statistical table of body weight, waist circumference and body fat rate of volunteers before and after test
Note that: * Indicating significant differences (P < 0.05) compared to the pre-test
As can be seen from the data in table 10, after 30 days of eating lactobacillus paracasei GF045 solid beverage, the test group women and men have significantly less body fat weight and significantly reduced body fat rate, wherein the female group body fat rate reaches 30.7% which is close to 30% of the standard of women participating in weight-losing test in the evaluation method described in "nine, contribution to control of in vivo fat under item" human body test food test "in" health food function test and evaluation method (2022 edition); after 60d consumption, the female body fat rate of the test group is reduced to 27.5% to reach the normal range, the weight loss is successful, and the male body fat rate is reduced to 24.8% which is slightly lower than the weight loss test standard male by more than 25%, and the weight loss is within the normal range. From test data, the female has obvious weight-reducing effect compared with the male, and can reach the normal range within 30 days, while the male can reach the normal standard within 60 days.
EXAMPLE 2 use of Lactobacillus paracasei GF045 in weight loss aid products
In view of the weight-losing efficacy of reducing the weight, the body fat percentage and the like of the lactobacillus paracasei GF045, and the indexes of reducing the total cholesterol, the triglyceride, the low-density lipoprotein cholesterol and the like in the blood fat, the lactobacillus paracasei GF045 can be used as the raw material of food, health-care food or medicine, and is further used for preparing the food, the health-care food or the medicine with auxiliary weight-losing according to the conventional application method of probiotics in the prior art.
EXAMPLE 3 Lactobacillus paracasei GF045 fermented sour meat
The present example provides a method for preparing fermented sour meat using lactobacillus paracasei GF045, and the physical and chemical indexes of the obtained sour meat were measured, and the obtained sour meat was subjected to sensory evaluation, and in addition, was compared with sour meat prepared according to the existing standard preparation method, and the specific experimental method is as follows.
(1) Method for fermenting sour meat by using lactobacillus paracasei GF045
The embodiment provides a method for fermenting sour meat by using lactobacillus paracasei GF045, which specifically comprises the following steps in sequence:
1000g of commercially available fresh streaky pork was washed and cut into 3cm X5 cm pieces;
taking 110g of millet, washing, adding the millet into a boiling pot filled with 2.2L of water, boiling with strong fire, regulating the small fire to keep slight boiling for 30min, measuring that the specific gravity of the juice is 1.05g/mL at 37 ℃, removing the millet to obtain 1.9L of millet juice, and adding 19g of glucose into the millet juice to prepare a solution A;
adding cut pork slices into the solution A, sealing, sterilizing at 80 ℃ for 3min, inoculating lactobacillus paracasei GF045 seed solution according to the inoculation amount of 3% of the volume of the millet juice, fermenting at 37 ℃ for 5 days, and obtaining the sour meat, namely the lactobacillus paracasei GF045 sour meat.
Wherein the viable count of the lactobacillus paracasei GF045 seed liquid is 5.0X10 9 CFU/mL；
During fermentation, a proper amount of pork is taken every day to detect fat, cholesterol and viable count, and meanwhile, the pH value of the millet juice is detected. In the embodiment, the fat detection adopts the method of 'GB 5009.6-2016 national food safety standard for determination of fat in food'; the cholesterol detection adopts GB 5009.128-2016 determination of cholesterol in national food safety standard food; the live bacteria detection of the lactobacillus paracasei GF045 adopts the lactobacillus detection of the national food safety standard food microbiology inspection of GB 4789.35-2016.
Specific statistics of the pH values of the sour meat fat, cholesterol and viable count and millet juice at different fermentation times are shown in Table 11.
TABLE 11 statistics of pH of sour meat fat, cholesterol and viable count and millet juice at various fermentation times
As can be seen from the data in table 11, the fresh pork, which is 50% fat and lean, was immersed and fermented in the millet juice by lactobacillus paracasei GF045, and the fat and cholesterol in the fresh pork were gradually degraded over the fermentation time, the degradation rates of the fat and cholesterol were gradually increased in the first 3 days, and the degradation rates were decreased in the 4 th and 5 th days; the viable count of Lactobacillus paracasei GF045 increases in pairs during the first 3 days, and enters the stationary phase during 4 and 5 days. The fat content and cholesterol content of the final sour meat were reduced by 42.3% and 35.1%, respectively, compared to fresh pork, the fat in the fresh meat was gradually catabolized by lactobacillus paracasei GF045.
(2) Sensory evaluation
The sensory evaluation of the lactobacillus paracasei GF045 sour meat prepared as described above was also carried out in this example, with the following specific results.
The evaluation of the results is mainly carried out by people of different sexes and different ages. The age is between 20 and 55, and the proportion of men and women is 1:1 and a total of 30 people make up a sensory panel.
The sensory scoring criteria were: the sour meat taste has pleasant fermented taste and the acidity is proper to be 5 minutes, and the lighter the fermented taste or the more sour the taste, the lower the score;
the degree of sour meat greasy satisfaction was scored, with a score of 5 points being full, with lower scores being more greasy.
The test was divided into a control group and a test group, wherein the control group was commercial fresh meat and the test group was Lactobacillus paracasei GF045 sour meat. On the premise of not adding edible oil, the fermented sour meat is salted for 30min by adopting a proper amount of salt, green onion, ginger, spice, soy sauce and vinegar, and simultaneously, the commercial fresh pork with five flowers is salted for 30min by adopting the same ingredients. The pickled sour meat and fresh meat are put into a steamer for steaming, steamed for 1 hour with big fire after being steamed, and the taste of the steamed sour meat and fresh meat is evaluated.
Sensory evaluation panels were made by sensory panel evaluation and statistics as shown in table 12 below.
Table 12 fresh meat and sour meat feel evaluation table
As can be seen from the data in table 12: the degree of greasiness of fresh meat fermented by lactobacillus paracasei GF045 is greatly reduced, people are satisfied with the degree of greasiness of the fermented pork, a small number of people can not eat meat acid, and knowledge explanation of fermented sour meat products is needed. Experimental data also show that the content of the fat in the sour meat fermented by the lactobacillus paracasei GF045 is reduced, and the fat intake of a human body can be reduced by taking the sour meat, so that the physical health is ensured.
(3) Comparison with the conventional method for preparing sour meat
In this example, the lactobacillus paracasei GF045 sour meat prepared as described above was compared with the sour meat prepared by the conventional method, and whether the sour meat obtained by the preparation method of the present invention is significantly different from the sour meat prepared by the conventional method in fat content and cholesterol content was examined, and the specific method is as follows.
Firstly, the sour meat of the traditional process is prepared according to the method recorded in the ' 7 th part of the sour fish of Hunan Carnis Pseudosciaenae ' of DB 43/T1589.7-2019 Hunan nationality dish '. 1000g of streaky pork with the same fat content as that selected in the method for fermenting sour meat by using lactobacillus paracasei GF045 in the embodiment (1) is cut into meat slices with the length of 7cm, the width of 5cm and the thickness of 3mm, 150g of salt, 8g of pricklyash peel powder and a proper amount of onion ginger cooking wine are added, and the mixture is uniformly stirred for pickling for 30-60 minutes; and then 500g pork: mixing pork and glutinous rice flour at a ratio of 200g corn flour (or glutinous rice flour), pressing in a jar, sealing with rice straw or husk, and fermenting in a water bowl for 20 days to obtain sour meat.
After the fermentation, the fat and cholesterol contents of the sour meat and the lactobacillus paracasei GF045 sour meat of the conventional process were measured, wherein the method for measuring the fat and cholesterol contents was the same as that described in the section of the present example (1). The content of fat and cholesterol in the two sour meats was determined as shown in table 13 below.
Table 13 statistics of fat and cholesterol levels in two sour meats
From the data in Table 13, the content of fat and cholesterol in the Lactobacillus paracasei GF045 sour meat is significantly lower than in the traditional process sour meat. The preparation method provided by the invention can be used for fermenting for 5 days to obtain the sour meat with proper sense and relatively low fat and cholesterol content, and the traditional process needs to be used for fermenting for more than 15 days when preparing the sour meat, and the fat and cholesterol content of the obtained sour meat is obviously higher than that of lactobacillus paracasei GF045 sour meat. Therefore, the preparation method provided by the invention can obviously improve the working efficiency, and the prepared product is healthier.
Examples 4 to 9 preparation method of sour meat
Examples 4 to 9 are a method for producing sour meat using Lactobacillus paracasei GF045, respectively, which are substantially the same as the production method described in section (1) of example 3, except that the control parameters are different, and the list of control parameters for the different examples is shown in Table 14.
Table 14 control parameter lists in examples 4 to 9
Claims (10)
1. A lactobacillus paracasei GF045 with the function of assisting weight losing is characterized in that the strain is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and the preservation number is CGMCC No.26482 and Latin name isLactobacillus paracasei。
2. Lactobacillus paracasei GF045 with weight loss aiding effect according to claim 1, characterized in that its gene sequence of 16SrDNA is as follows:
GAGTTCTCCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGATCAGGTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCTAGTAAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTTTT。
3. use of lactobacillus paracasei GF045 with weight loss aiding effect according to claim 1 or 2 for the preparation of food, health food or pharmaceutical products with weight loss aiding effect.
4. Use of lactobacillus paracasei GF045 with weight loss aiding effect according to claim 1 or 2 for the preparation of sour meat.
5. The method for producing sour meat according to claim 4, comprising the steps of:
placing pork in a solution A containing glucose and millet juice, sealing, sterilizing, inoculating lactobacillus paracasei GF045, and fermenting until the pH is 4.3-4.5 to obtain the sour meat.
6. The method for preparing sour meat according to claim 5, wherein the mass ratio of glucose to millet juice in the solution a is 1: 50-100, wherein the mass ratio of pork to the solution A is 1: 2-2.5.
7. The method for preparing sour meat according to claim 6, wherein the method for preparing millet juice comprises adding millet into water and decocting, stopping decocting when the specific gravity of the juice is within the range of 1.05-1.35 g/mL at 35-37 ℃, and removing the millet to obtain the millet juice.
8. The method for producing sour meat according to claim 5, wherein the sterilization temperature is 75-80 ℃ and the time is 3-5 min.
9. The method for producing sour meat according to any one of claims 5 to 8, wherein the viable count of lactobacillus paracasei GF045 is 4.0×10 9 ～8.0×10 9 cfu/mL, the inoculation amount is 3-10%.
10. The method for producing sour meat according to claim 9, wherein the fermentation time is 72 to 120 hours and the fermentation temperature is 35 to 37 ℃.
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