CN109971668B - Lactobacillus plantarum with cholesterol reducing function and application and microecological preparation thereof - Google Patents
Lactobacillus plantarum with cholesterol reducing function and application and microecological preparation thereof Download PDFInfo
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- CN109971668B CN109971668B CN201910029597.5A CN201910029597A CN109971668B CN 109971668 B CN109971668 B CN 109971668B CN 201910029597 A CN201910029597 A CN 201910029597A CN 109971668 B CN109971668 B CN 109971668B
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- lactobacillus plantarum
- cholesterol
- lactobacillus
- preparation
- reducing function
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Abstract
The invention relates to the field of microorganisms, and particularly provides lactobacillus plantarum with a cholesterol-reducing function, application thereof and a microecological preparation. The lactobacillus is obtained by separating from the intestinal tract of healthy adult minks, is identified and named as lactobacillus plantarum S2303, and has a preservation number of CCTCC M2018941. The lactobacillus plantarum has good genetic stability, researches show that the lactobacillus plantarum S2303 has a remarkable degradation effect on cholesterol, the degradation rate of the cholesterol can reach 82.22% in the presence of cholate, and the lactobacillus plantarum S2303 also has a good cholesterol-reducing function in organisms.
Description
Technical Field
The invention relates to the field of microorganisms, and particularly relates to lactobacillus plantarum with a cholesterol-reducing function, application thereof and a microecological preparation.
Background
Lactic acid bacteria (lactic acid bacteria) are ubiquitous and closely related to human health. Lactic acid bacteria are widely present in the digestive system, respiratory system, oral cavity, skin and feces of humans and animals. In a healthy state, lactic acid bacteria have a plurality of excellent probiotic functions of inhibiting pathogenic bacteria, improving the intestinal environment, regulating the intestinal flora, improving the growth performance and the like, and are important microorganisms widely applied to food, medicines and feed additives. In recent years, as research on lactic acid bacteria has become more widespread, many functions of lactic acid bacteria have become known. Research shows that the probiotics comprising the lactic acid bacteria have the effects of increasing the weight gain of young livestock and poultry, improving the digestibility of nutrient substances, improving the reward of feed, reducing the diarrhea rate, relieving the stress condition, reducing the death rate and the like. Meanwhile, part of the lactic acid bacteria can adjust abnormal metabolism to restore the normal metabolism, and can also directly participate in cholesterol metabolism and the like. Research also shows that the lactobacillus has obvious immunity promotion effect on organisms, can promote the development of intestinal immune systems of newborns, can also promote local and systemic immune functions of intestinal mucosa, has regulation effect on both specific and non-specific immune responses, maintains the normal level of the immune functions of the organisms in a healthy state, and the like.
Lactic acid bacteria that have been found in nature at present are classified into 43 genera including lactobacillus, lactococcus, enterococcus, streptococcus, pediococcus, leuconostoc, and the like in terms of bacteriology. The Lactobacillus species mainly relate to Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus plantarum and the like.
Although lactic acid bacteria have many of the above probiotic effects, not only are species of the lactic acid bacteria significantly different in probiotic effect, but also the strains of different habitats are significantly different from one another. The culture time, the additive amount and the like have different effects on the application.
Elevated cholesterol levels are one of the major risk factors for cardiovascular disease. An important probiotic function of lactic acid bacteria is the direct involvement in the metabolism of cholesterol. The lactic acid bacteria entering the body through the digestive system must be able to withstand the strong acid and high bile salt environment of the digestive tract and exert a probiotic effect on the body, but the degradation capability of the existing lactic acid bacteria on cholesterol is still to be further improved.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to provide lactobacillus plantarum S2303 which has remarkable capacity of degrading cholesterol.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
lactobacillus plantarum S2303 with preservation number of CCTCC M2018941.
The Lactobacillus plantarum S2303 can be used for regulating intestinal microecological balance.
The Lactobacillus plantarum S2303 can be used for preparing products for regulating intestinal microecological balance.
Further, the product comprises a probiotic.
A microecological preparation contains Lactobacillus plantarum S2303 as effective component.
Use of the lactobacillus plantarum S2303 described above for the reduction of cholesterol for non-therapeutic purposes.
The lactobacillus plantarum S2303 is applied to preparation of products with cholesterol reducing function.
Further, the product comprises any one of medicines, cosmetics, foods, health products, leavening agents, animal feeds or pet foods;
preferably, the food product comprises yoghurt, fermented milk, cream, cheese, drink, sausage or bread.
The lactobacillus plantarum S2303 is applied to the breeding industry.
Further, the breeding industry includes broiler production, pig fattening, cattle fattening, piglet production or cow production.
Compared with the prior art, the invention has the beneficial effects that:
the lactobacillus is obtained by separating from the intestinal tract of healthy adult minks, is identified and named as lactobacillus plantarum S2303, and has a preservation number of CCTCC M2018941. The lactobacillus plantarum has good genetic stability, researches find that the lactobacillus plantarum S2303 has obvious degradation effect on both yolk cholesterol and skim milk cholesterol, the degradation rate of the cholesterol can reach 82.22% in the presence of cholate, and a mouse serum cholesterol content degradation test also proves that the lactobacillus plantarum S2303 also has good cholesterol-reducing function in organisms. The lactobacillus plantarum S2303 provided by the invention has strong and stable capacity of reducing cholesterol, has good probiotic characteristics, can be used for developing a cholesterol-reducing probiotic product or fermenting and preparing a product with low cholesterol content, and has wide application prospects in production and life.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a scanning electron micrograph of Lactobacillus plantarum S2303 provided by the present invention in example 2;
FIG. 2 is a diagram showing the results of gene sequence alignment analysis of the 1 st 16S rRNA of Lactobacillus plantarum S2303 according to the present invention in example 3;
FIG. 3 is a diagram showing the results of gene sequence alignment analysis of the 7 th 16S rRNA of Lactobacillus plantarum S2303 according to the present invention in example 3;
FIG. 4 is a diagram showing the results of alignment analysis of the 1 st pheS gene sequences of Lactobacillus plantarum S2303 according to the present invention in example 3;
FIG. 5 is a diagram showing the results of comparison and analysis of the 7 th-generation pheS gene sequences of Lactobacillus plantarum S2303 according to the present invention in example 3;
FIG. 6 shows the degradation rate of Lactobacillus plantarum S2303 provided by the present invention to cholesterol in egg yolk under in vitro culture conditions in example 4, with the abscissa representing the culture time (unit: hour) and the ordinate representing the degradation rate (%);
FIG. 7 shows the degradation rate of the Lactobacillus plantarum S2303 provided by the invention on skim milk cholesterol under in vitro culture conditions in example 5;
FIG. 8 shows the degradation rate of Lactobacillus plantarum S2303 provided by the present invention on cholesterol in the presence of cholate in vitro culture in example 6;
FIG. 9 shows the effect of Lactobacillus plantarum S2303 provided by the invention in example 7 on serum cholesterol in mice.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer.
In the present invention, all the embodiments and preferred methods mentioned herein can be combined with each other to form a new technical solution, if not specifically stated.
In the present invention, all the technical features mentioned herein and preferred features may be combined with each other to form a new technical solution, if not specifically stated.
In the present invention, the components referred to or the preferred components thereof may be combined with each other to form a novel embodiment, if not specifically stated.
The "ranges" disclosed herein may have one or more lower limits and one or more upper limits, respectively, in the form of lower limits and upper limits.
In the present invention, unless otherwise specified, the individual reactions or operation steps may or may not be performed in sequence. Preferably, the reaction processes herein are carried out sequentially.
Unless otherwise defined, technical and scientific terms used herein have the same meaning as is familiar to those skilled in the art. In addition, any methods or materials similar or equivalent to those described herein can also be used in the present invention.
The lactobacillus plantarum S2303 is preserved in China Center for Type Culture Collection (CCTCC) in 2018, 12 months and 28 days, the preservation unit address is eight-way Lopa nationality mountain in Wuchang district, Wuhan City, Hubei province, the preservation number is CCTCC M2018941, the classification name of the strain is lactobacillus plantarum, the latin name is: lactobacillus plantarum.
Minks, which belong to the family of the weasels, meat animals, can tolerate a daily ration with a fat content of up to 35%, even though normally the fat content in mink diets averages around 25% (cumulative statistics in the present study). The lactobacillus plantarum S2303 is obtained by screening lactobacillus capable of degrading cholesterol from mink sources, evaluating the biological characteristics and the degradation effect of the lactobacillus on cholesterol and detecting the advantageous effects of the lactobacillus plantarum based on the intestinal microbiology which plays a decisive role in nutrient digestion as a main entry point. The lactobacillus plantarum has good genetic stability, researches find that the lactobacillus plantarum S2303 has obvious degradation effect on both yolk cholesterol and skim milk cholesterol, the degradation rate of the cholesterol can reach 82.22% in the presence of cholate, and a mouse serum cholesterol content degradation test also proves that the lactobacillus plantarum S2303 also has good cholesterol-reducing function in organisms. Shows that the lactobacillus plantarum S2303 provided by the invention
Has strong and stable capacity of reducing cholesterol and good probiotic characteristics, can be used for developing probiotic products for reducing cholesterol or fermenting products for preparing products with low cholesterol content, and has wide application prospect in production and life.
The specific process for screening and obtaining the lactobacillus plantarum S2303 comprises the following steps: separating lactobacillus from intestinal tract of healthy adult mink, adding physiological saline into the content of the distal end of small intestine, shaking, mixing, performing gradient dilution to obtain gradient dilution, performing separation culture on the gradient dilution by using a pouring culture method, and purifying the strain by using a plate-streaking method. Inoculating the purified lactobacillus into an MRS liquid culture medium, detecting the microbiological characteristics of the lactobacillus, such as morphology, physiological characteristics, sugar catabolism capability and morphology of the lactobacillus by scanning electron microscope observation, detecting the degradation function of the lactobacillus to cholesterol by in vitro culture, and detecting the safety of mice and serum cholesterol removal capability to obtain the lactobacillus plantarum.
The invention provides application of the lactobacillus plantarum S2303 in regulating intestinal microecological balance and application in preparing a product for regulating intestinal microecological balance. Lactobacillus plantarum S2303 is used as probiotic bacteria, and has effects in promoting health of consumer, inhibiting propagation of harmful bacteria in intestine, and regulating intestinal microecological balance. The product prepared for regulating intestinal microecological balance, such as but not limited to microecologics, has good effect.
A microecological preparation contains Lactobacillus plantarum S2303 as effective component.
A microecological preparation (Probiotics), also called a live bacterial preparation (Biogen) or a biological agent, is a preparation prepared by using Probiotics beneficial to and harmless to a host or growth promoting substances of the Probiotics through a special process by applying the microecological principle. Wherein the characteristic bacteria must exist in a large amount, and can continue to survive and have activity.
The lactobacillus plantarum S2303 of the present invention is used for cholesterol reduction for non-therapeutic purposes.
The lactobacillus plantarum S2303 is applied to preparation of products with the function of reducing cholesterol.
The product can be any one of medicines, cosmetics, foods, health products, leavening agents, animal feeds or pet foods.
The food may be, but is not limited to, yogurt, fermented milk, cream, cheese, drink, sausage, or bread.
The lactobacillus plantarum S2303 can be applied to the breeding industry. For example, but not limited to broiler production, fattening pig, fattening cattle, piglet production or cow production.
The invention is further illustrated by the following specific examples, which, however, are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.
Example 1 screening of Lactobacillus plantarum S2303
Adding 20g of the content of the far end of small intestine of healthy adult mink into 180ml of sterile water, shaking in shaking table for 30min to dissolve the particulate matter sufficiently, according to the ratio of 0, 102、103、104、105、106、107、108And 109Diluting with 10 gradients, performing isolation culture by using a pouring culture method, culturing at 37 deg.C for 72h, selecting colony with calcium-dissolving ring, bulge, whole edge and white color, purifying by plate-streaking method for 3-4 generations, and obtaining pure Lactobacillus plantarum S2303 biological culture.
Example 2 identification of Lactobacillus plantarum S2303
Inoculating the purified Lactobacillus plantarum S2303 into an MRS liquid culture medium, and detecting the microbiological characteristics, such as morphological characteristics and physiological characteristics.
Morphological characteristics
Culturing in MRS culture medium at 37 deg.C for 72 hr to obtain rod-shaped thallus of 0.4 μm × 1.2-1.9 μm, single or fence-shaped thallus, gram-positive thallus, white and round shape, and has convex surface, moist, opaque, and neat edge. As shown in FIG. 1, the scanning results of the electron microscope showed that the cells were rod-shaped, 0.6-0.8. mu. m.times.0.9-1.6. mu.m, and had fine particles on the surface.
Physiological characteristics
The physiological and biochemical characteristics of lactobacillus plantarum S2303 were detected, and the results are shown in table 1:
TABLE 1 physiological and biochemical characteristics of Lactobacillus plantarum S2303
The TLC analysis results are: the main amino acid composition of the cell wall of the lactobacillus plantarum S2303 provided by the invention is Mesco-DAP (diaminopimelic acid), alanine, glutamic acid and glycine; the main sugar component of the whole-cell hydrolysate is galactose.
The HPLC analysis result is: the (G + C) mol% content was 45.3 mol%.
Example 3 genetic stability analysis of Lactobacillus plantarum S2303
Continuously subculturing the lactobacillus plantarum S2303 slant for 7 generations, and respectively carrying out morphological observation, physiological and biochemical characteristic detection, 16S rRNA sequence sequencing and comparative analysis, and pheS gene sequence sequencing and gene sequence comparative analysis on strains of the 1 st generation and the 7 th generation.
The 1 st generation 16S rDNA sequence is as follows (SEQ ID NO. 1):
GTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAAC
the 7 th 16S rDNA sequence is as follows (SEQ ID NO. 2):
TGCTATACATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAAC
the 1 st pheS sequence is as follows (SEQ ID NO. 3):
CGTGACATGCAAGACACGTTCTATATTACCAAAGACGTGCTACTACGCACGCAGACGTCTGCTGATCAGCCGCGGTCACTTGAAAATCACGATTTTTCTAAAGGACCGCTGAAGGTCTTGTCACCTGGCCGCGTTTATCGGCGTGATACGGATGATGCAACCCATTCCCATCAATTTCATCAAATTGAAGGGTTAGTCGTGGACAAGCATATTACGATGGCTGATTTGAAGGGCACCTTAATTCTGGTTGCCAAGACTTTGTTTGGCGATCAATTCGATGTTCGGCTACGGCCAAGCTTCTTTCCATTCACGGAACCATCCGTAGAAGCTGATGTAACTTGCTTTAATTGCAATGGCAAGGGCTGTGCAATCTGTAAGCAAACGGGTTGGATCGAAGTACTGGGTGCCGGCATGGTTCACCCCCACGTGTTAGAAATGTCTGGCATTGATCCAGAAGAATATGGTGGCTTTGCTT
the 7 th pheS sequence is as follows (SEQ ID NO. 4):
GACATGCAAGACACGTTCTATATTACCAAAGACGTGCTACTACGCACGCAGACGTCTGCTGATCAGCCGCGGTCACTTGAAAATCACGATTTTTCTAAAGGACCGCTGAAGGTCTTGTCACCTGGCCGCGTTTATCGGCGTGATACGGATGATGCAACCCATTCCCATCAATTTCATCAAATTGAAGGGTTAGTCGTGGACAAGCATATTACGATGGCTGATTTGAAGGGCACCTTAATTCTGGTTGCCAAGACTTTGTTTGGCGATCAATTCGATGTTCGGCTACGGCCAAGCTTCTTTCCATTCACGGAACCATCCGTAGAAGCTGATGTAACTTGCTTTAATTGCAATGGCAAGGGCTGTGCAATCTGTAAGCAAACGGGTTGGATCGAAGTACTGGGTGCCGGCATGGTTCACCCCCACGTGTTAGAAATGTCTGGCATTGATCCAGAAGAATATGGTGGTTTTGC
the results of the 1 st generation lactobacillus plantarum S2303 gene sequence alignment analysis based on 16S rRNA are shown in fig. 2, and the results of the gene sequence alignment analysis based on pheS are shown in fig. 4. Through Blast comparison, the sequence homology of the 16S rRNA gene sequence and the pheS gene sequence with lactobacillus plantarum reaches 100%. The results of the alignment analysis of the Lactobacillus plantarum S2303 generation 7 based on the 16S rRNA gene sequence are shown in FIG. 3, and the results of the alignment analysis based on the pheS gene sequence are shown in FIG. 5. Through Blast comparison, the sequence homology of the 16S rRNA gene sequence and the pheS gene sequence with lactobacillus plantarum reaches 100%. Through the experiment, the following results are found: the morphological characteristics of the strain of the lactobacillus plantarum S2303 are not changed, the physiological and biochemical characteristics are not changed obviously, the 16S rRNA gene sequence is not changed, the pheS gene sequence is not changed, and the strain character can be inherited stably.
Example 4 degradation of egg yolk cholesterol by Lactobacillus plantarum S2303
Adopts 100ml of MRS basic culture medium, adds fresh egg yolk, the amount of the egg yolk is about 4 percent of the amount of the culture medium, and carefully removes the egg yolk by using a 1ml sterilization pipette. Inoculating 100ul of fresh bacteria liquid cultured for 18h into the mixed culture medium, culturing at 37 ℃, taking 10ml of culture solution in 4, 8, 12, 16 and 24h respectively, adding into a sterilized centrifuge tube with a cover, storing at 4 ℃, immediately centrifuging at 6000rpm/min for 15min after all samples are taken, taking supernatant, and performing colorimetric detection at 550nm by using an o-phthalaldehyde method. The degradation rate of egg yolk cholesterol was counted, and the results are shown in fig. 6.
The cholesterol content of 1 common egg is about 250mg, the daily intake of cholesterol of each person is not higher than 300mg, and excessive intake of cholesterol can cause atherosclerosis, thereby inducing cardiovascular and cerebrovascular diseases such as cerebral infarction, coronary heart disease and the like. In the experiment, the yolk containing higher cholesterol in nature is taken as an experimental material, the influence of lactobacillus plantarum S2303 on the in-vitro degradation of egg cholesterol is researched, and basic data are provided for the utilization of lactobacillus plantarum S2303.
The cholesterol content in the egg yolk is 13.89 mg/g. As can be seen from FIG. 6, the Lactobacillus plantarum S2303, when cultured in MRS-egg yolk medium for 24h, has a high cholesterol removal rate from egg yolk without cholate in the initial stage, which is up to 63.07% at 4h, and gradually decreases the cholesterol removal rate with the increase of the culture time, and remains substantially stable after 16h, which is about 10%.
Example 5 degradation of skim milk Cholesterol by Lactobacillus plantarum S2303
A skim milk fermentation medium with skim milk content of 12% and glucose content of 8% was used, and after 24 hours of culture, sampling and centrifugation were performed, and the assay was the same as in example 4. The results are shown in FIG. 7.
The content of cholesterol in the skim milk powder is 28mg/100 mg. As can be seen from FIG. 7, under the in vitro culture conditions, Lactobacillus plantarum S2303 has a degradation effect on cholesterol in natural skim milk, and the degradation rate reaches 22.35% after 24h of culture. It is demonstrated that the lactobacillus plantarum S2303 provided by the present invention can utilize cholesterol even in the case of low cholesterol content, and at the same time, it is demonstrated that lactobacillus plantarum S2303 can grow in skim milk and has the ability to be applied to yogurt fermentation production.
Example 6 degradation of cholesterol by Lactobacillus plantarum S2303 in the Presence of cholate
10mg/ml of sterile cholesterol (purity > 99%, sigma) solution was added to the culture medium to a final concentration of 70. mu.g/ml, while 2% of bovine cholate (2 g of bovine cholate added to 1L of medium) was added to the culture medium, and Lactobacillus plantarum S2303 was inoculated in an inoculum size of 1%, cultured at 37 ℃ for 24 hours in the same manner as in example 4, and the results are shown in FIG. 8. Under the condition of the existence of cholate, the degradation rate of the lactobacillus plantarum S2303 to cholesterol is 82.22%, which is better than 81% and 72% in the prior art.
Example 7 Effect of Lactobacillus plantarum S2303 on serum cholesterol levels in mice
Feeding 45d Kunming mice in cages, dividing into 2 groups, control group and S2303, centrifuging fresh liquid test strain cultured daily at 5000rpm/min before each administration, centrifuging for 15min, collecting thallus precipitate, diluting with normal saline, and making into cell suspension 0.3ml per mouse with viable bacteria content of about 3 × 1011cfu/ml, the control group was administered with an equal amount of physiological saline, and administered orally with a gastric lavage apparatus. Each group had 15 animals, and the rats were fed with free water. And (30) limiting feeding for 12h for each group of mice, removing eyeballs in the morning to collect blood, immediately centrifuging all blood samples at 6000rpm/min for 10 minutes, taking serum, and measuring the content of total cholesterol in the serum by using a full-automatic biochemical analyzer.
Every 1% of cholesterol in plasma is reduced, and the risk of coronary heart disease is reduced by 3-4%. As shown in FIG. 9, the serum cholesterol level of Lactobacillus plantarum S2303 was reduced by 0.26mmol/L and the serum cholesterol reduction rate was 18.71% in the S2303 group, both of which were drenched and administered without death or abnormality. The lactobacillus plantarum S2303 provided by the invention has the effect of reducing the total cholesterol content in the serum of a mouse, and compared with other lactic acid bacteria, the lactobacillus plantarum has the cholesterol degradation capability which is less seen in other habitats.
While particular embodiments of the present invention have been illustrated and described, it would be obvious that various other changes and modifications can be made without departing from the spirit and scope of the invention. It is therefore intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.
SEQUENCE LISTING
<110> institute of specialty products of Chinese academy of agricultural sciences
<120> Lactobacillus plantarum with cholesterol-reducing function, application thereof and microecological preparation
<130> 2018
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 1412
<212> DNA
<213> Lactobacillus plantarum
<400> 1
gtcgaacgaa ctctggtatt gattggtgct tgcatcatga tttacatttg agtgagtggc 60
gaactggtga gtaacacgtg ggaaacctgc ccagaagcgg gggataacac ctggaaacag 120
atgctaatac cgcataacaa cttggaccgc atggtccgag tttgaaagat ggcttcggct 180
atcacttttg gatggtcccg cggcgtatta gctagatggt ggggtaacgg ctcaccatgg 240
caatgatacg tagccgacct gagagggtaa tcggccacat tgggactgag acacggccca 300
aactcctacg ggaggcagca gtagggaatc ttccacaatg gacgaaagtc tgatggagca 360
acgccgcgtg agtgaagaag ggtttcggct cgtaaaactc tgttgttaaa gaagaacata 420
tctgagagta actgttcagg tattgacggt atttaaccag aaagccacgg ctaactacgt 480
gccagcagcc gcggtaatac gtaggtggca agcgttgtcc ggatttattg ggcgtaaagc 540
gagcgcaggc ggttttttaa gtctgatgtg aaagccttcg gctcaaccga agaagtgcat 600
cggaaactgg gaaacttgag tgcagaagag gacagtggaa ctccatgtgt agcggtgaaa 660
tgcgtagata tatggaagaa caccagtggc gaaggcggct gtctggtctg taactgacgc 720
tgaggctcga aagtatgggt agcaaacagg attagatacc ctggtagtcc ataccgtaaa 780
cgatgaatgc taagtgttgg agggtttccg cccttcagtg ctgcagctaa cgcattaagc 840
attccgcctg gggagtacgg ccgcaaggct gaaactcaaa ggaattgacg ggggcccgca 900
caagcggtgg agcatgtggt ttaattcgaa gctacgcgaa gaaccttacc aggtcttgac 960
atactatgca aatctaagag attagacgtt cccttcgggg acatggatac aggtggtgca 1020
tggttgtcgt cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga gcgcaaccct 1080
tattatcagt tgccagcatt aagttgggca ctctggtgag actgccggtg acaaaccgga 1140
ggaaggtggg gatgacgtca aatcatcatg ccccttatga cctgggctac acacgtgcta 1200
caatggatgg tacaacgagt tgcgaactcg cgagagtaag ctaatctctt aaagccattc 1260
tcagttcgga ttgtaggctg caactcgcct acatgaagtc ggaatcgcta gtaatcgcgg 1320
atcagcatgc cgcggtgaat acgttcccgg gccttgtaca caccgcccgt cacaccatga 1380
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<213> Lactobacillus plantarum
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acacctggaa acagatgcta ataccgcata acaacttgga ccgcatggtc cgagtttgaa 180
agatggcttc ggctatcact tttggatggt cccgcggcgt attagctaga tggtggggta 240
acggctcacc atggcaatga tacgtagccg acctgagagg gtaatcggcc acattgggac 300
tgagacacgg cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa 360
agtctgatgg agcaacgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt 420
taaagaagaa catatctgag agtaactgtt caggtattga cggtatttaa ccagaaagcc 480
acggctaact acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt 540
attgggcgta aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggctcaa 600
ccgaagaagt gcatcggaaa ctgggaaact tgagtgcaga agaggacagt ggaactccat 660
gtgtagcggt gaaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctgg 720
tctgtaactg acgctgaggc tcgaaagtat gggtagcaaa caggattaga taccctggta 780
gtccataccg taaacgatga atgctaagtg ttggagggtt tccgcccttc agtgctgcag 840
ctaacgcatt aagcattccg cctggggagt acggccgcaa ggctgaaact caaaggaatt 900
gacgggggcc cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct 960
taccaggtct tgacatacta tgcaaatcta agagattaga cgttcccttc ggggacatgg 1020
atacaggtgg tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca 1080
acgagcgcaa cccttattat cagttgccag cattaagttg ggcactctgg tgagactgcc 1140
ggtgacaaac cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg 1200
ctacacacgt gctacaatgg atggtacaac gagttgcgaa ctcgcgagag taagctaatc 1260
tcttaaagcc attctcagtt cggattgtag gctgcaactc gcctacatga agtcggaatc 1320
gctagtaatc gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc 1380
ccgtcacacc atgagagttt gtaacaccca aagtcggtgg ggtaac 1426
<210> 3
<211> 475
<212> DNA
<213> Lactobacillus plantarum
<400> 3
cgtgacatgc aagacacgtt ctatattacc aaagacgtgc tactacgcac gcagacgtct 60
gctgatcagc cgcggtcact tgaaaatcac gatttttcta aaggaccgct gaaggtcttg 120
tcacctggcc gcgtttatcg gcgtgatacg gatgatgcaa cccattccca tcaatttcat 180
caaattgaag ggttagtcgt ggacaagcat attacgatgg ctgatttgaa gggcacctta 240
attctggttg ccaagacttt gtttggcgat caattcgatg ttcggctacg gccaagcttc 300
tttccattca cggaaccatc cgtagaagct gatgtaactt gctttaattg caatggcaag 360
ggctgtgcaa tctgtaagca aacgggttgg atcgaagtac tgggtgccgg catggttcac 420
ccccacgtgt tagaaatgtc tggcattgat ccagaagaat atggtggctt tgctt 475
<210> 4
<211> 470
<212> DNA
<213> Lactobacillus plantarum
<400> 4
gacatgcaag acacgttcta tattaccaaa gacgtgctac tacgcacgca gacgtctgct 60
gatcagccgc ggtcacttga aaatcacgat ttttctaaag gaccgctgaa ggtcttgtca 120
cctggccgcg tttatcggcg tgatacggat gatgcaaccc attcccatca atttcatcaa 180
attgaagggt tagtcgtgga caagcatatt acgatggctg atttgaaggg caccttaatt 240
ctggttgcca agactttgtt tggcgatcaa ttcgatgttc ggctacggcc aagcttcttt 300
ccattcacgg aaccatccgt agaagctgat gtaacttgct ttaattgcaa tggcaagggc 360
tgtgcaatct gtaagcaaac gggttggatc gaagtactgg gtgccggcat ggttcacccc 420
cacgtgttag aaatgtctgg cattgatcca gaagaatatg gtggttttgc 470
Claims (9)
1. Lactobacillus plantarum S2303 with preservation number of CCTCC M2018941.
2. Use of lactobacillus plantarum S2303 according to claim 1 for the preparation of a product for the regulation of intestinal microecological balance.
3. Use according to claim 2, wherein the product comprises a probiotic.
4. A microecological preparation which comprises the Lactobacillus plantarum S2303 according to claim 1 as an active ingredient.
5. Use of lactobacillus plantarum S2303 according to claim 1 for the preparation of a product with cholesterol-lowering function.
6. Use according to claim 5, wherein the product comprises any of a pharmaceutical, a cosmetic, a food product, a nutraceutical, a starter, an animal feed or a pet food.
7. Use according to claim 6, wherein the food product comprises yoghurt, fermented milk, cream, cheese, drink, sausage or bread.
8. Use of lactobacillus plantarum S2303 according to claim 1 for the preparation of products for aquaculture.
9. Use according to claim 8, wherein the breeding industry comprises broiler production, fattening pigs, fattening cattle, piglet production or dairy production.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102093966A (en) * | 2010-12-02 | 2011-06-15 | 中国农业科学院特产研究所 | Mink-derived Lactobacillus plantarum strain MDL1118 and application thereof |
| CN108728382A (en) * | 2018-06-07 | 2018-11-02 | 四川大学 | One plant of tool norcholesterol and lactobacillus plantarum and its application for promoting enteron aisle short chain fatty acids generation ability |
| CN110317757A (en) * | 2019-07-16 | 2019-10-11 | 自然资源部第三海洋研究所 | Lactobacillus plantarum HJ-S2 and its application of one plant of tool norcholesterol and selenium enriching functions |
-
2019
- 2019-01-11 CN CN201910029597.5A patent/CN109971668B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102093966A (en) * | 2010-12-02 | 2011-06-15 | 中国农业科学院特产研究所 | Mink-derived Lactobacillus plantarum strain MDL1118 and application thereof |
| CN108728382A (en) * | 2018-06-07 | 2018-11-02 | 四川大学 | One plant of tool norcholesterol and lactobacillus plantarum and its application for promoting enteron aisle short chain fatty acids generation ability |
| CN110317757A (en) * | 2019-07-16 | 2019-10-11 | 自然资源部第三海洋研究所 | Lactobacillus plantarum HJ-S2 and its application of one plant of tool norcholesterol and selenium enriching functions |
Non-Patent Citations (3)
| Title |
|---|
| Ability of lactic acid bacteria isolated from mink to remove cholesterol: in vitro and in vivo studies;Hanlu Liu,等;《Can. J. Microbiol》;20130620;第59卷;563-569 * |
| 不同乳酸杆菌添加剂对水貂生长性能、营养物质消化率、氮平衡及血清生化指标的影响;荆等;《动物营养学报》;20130915(第09期);全文 * |
| 益生菌益生特性的研究进展;刘晗璐等;《中国畜牧兽医》;20100220(第02期);全文 * |
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