CN111011866B - Application of prebiotics in promoting growth of lactobacillus plantarum in intestinal environment - Google Patents
Application of prebiotics in promoting growth of lactobacillus plantarum in intestinal environment Download PDFInfo
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- 230000001737 promoting effect Effects 0.000 title claims abstract description 21
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Images
Classifications
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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
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
-
- 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
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides application of prebiotics in promoting growth of lactobacillus plantarum in an intestinal environment, and belongs to the field of microorganisms. The invention provides an application of fructo-oligosaccharide or galacto-oligosaccharide in promoting growth of lactobacillus plantarum AR237 in an intestinal environment, and also provides an application of isomaltooligosaccharide in promoting proliferation of lactobacillus plantarum AR113 or lactobacillus plantarum AR237 in a middle fermentation stage. Not all prebiotics have a proliferation promoting effect on lactobacillus plantarum in the middle of fermentation, and some prebiotics also have a matrix inhibiting effect.
Description
Technical Field
The invention relates to application of prebiotics in promoting growth of lactobacillus plantarum in an intestinal environment, and belongs to the field of microorganisms.
Background
The prebiotics are difficult to be absorbed and utilized by human intestinal tracts, but can be selectively utilized by microorganisms as nutrient components, and can improve the metabolism of the probiotics and promote the growth and reproduction of the probiotics. Due to the important role of the micro-ecological environment of human intestinal microorganisms, prebiotics and probiotics are at the focus of research and many correlation studies are being carried out around them.
The lactobacillus plantarum is one of lactobacillus, has the optimal growth temperature of 30-35 ℃, is anaerobic or facultative anaerobic, has straight or bent rod-shaped strains, is single or sometimes paired or chained, has the optimal pH of about 6.5, and belongs to homofermentation lactobacillus. Lactobacillus plantarum has a number of health-care effects, as follows: (1) has certain immunoregulation function; (2) has an inhibiting effect on pathogenic bacteria; (3) reducing serum cholesterol levels and preventing cardiovascular disease; (4) maintaining the flora balance in the intestinal tract; (5) promoting the absorption of nutrient substances; (6) relieving lactose intolerance; (7) inhibiting the formation of tumor cells, etc.
However, different probiotics have extremely strong strain specificity, and the utilization of prebiotics by different probiotics and even different probiotic strains are different. In the prior art, the study of the lactobacillus plantarum on the utilization of the prebiotics is lacked, and particularly the study of the lactobacillus plantarum on the utilization of the prebiotics in the intestinal environment is lacked. Through research on the utilization of the lactobacillus plantarum on the prebiotics in the intestinal environment, people can develop lactobacillus beverage or food which has higher nutritive value and can condition the intestines and the stomach of a human body.
Disclosure of Invention
The present invention has been made to solve the above problems, and an object of the present invention is to provide a use of prebiotics for promoting the growth of lactobacillus plantarum in an intestinal environment.
The invention provides application of prebiotics in promoting the growth of lactobacillus plantarum in an intestinal environment, which is characterized by comprising the following components: the prebiotics are fructo-oligosaccharide or galacto-oligosaccharide.
In the application of the prebiotics provided by the invention in promoting the growth of lactobacillus plantarum in an intestinal environment, the prebiotics can also have the following characteristics: wherein the lactobacillus plantarum is lactobacillus plantarum AR237.
In the application of the prebiotics provided by the invention in promoting the growth of lactobacillus plantarum in an intestinal environment, the prebiotics can also have the following characteristics: wherein the concentration of the prebiotics is 10 mug/mL-30 mug/mL.
The invention also provides application of isomaltose hypgather in promoting proliferation of lactobacillus plantarum in the middle stage of fermentation.
In the application of the isomaltooligosaccharide provided by the invention in promoting the proliferation of lactobacillus plantarum in the middle stage of fermentation, the isomaltooligosaccharide also has the following characteristics: wherein the concentration of isomaltooligosaccharide is 10. Mu.g/mL-30. Mu.g/mL.
In the application of the isomaltooligosaccharide provided by the invention in promoting the proliferation of lactobacillus plantarum in the middle stage of fermentation, the isomaltooligosaccharide also has the following characteristics: wherein the Lactobacillus plantarum is Lactobacillus plantarum AR113 or Lactobacillus plantarum AR237.
Action and effects of the invention
According to the application of the prebiotics in the intestinal environment to promote the growth of the lactobacillus plantarum, the fructo-oligosaccharides and the galactooligosaccharides can be absorbed and utilized by specific lactobacillus plantarum strains in the intestinal microenvironment better, so that the number of specific lactobacillus plantarum is increased remarkably within a certain time, and therefore the fructo-oligosaccharides and the galactooligosaccharides provided by the invention can promote the growth of the specific lactobacillus plantarum.
Drawings
FIG. 1 is a graph of the growth of Lactobacillus plantarum AR113 in a medium with different prebiotics as carbon source in an example of the invention;
FIG. 2 is a graph of the growth of Lactobacillus plantarum AR117 in a medium with different prebiotics as carbon source in an example of the invention;
FIG. 3 is a graph of the growth of Lactobacillus plantarum AR237 in a medium with different prebiotics as carbon source in an example of the invention;
FIG. 4 is a graph of the growth of Lactobacillus plantarum AR509 in a medium with different prebiotics as carbon source in an example of the invention;
FIG. 5 is a graph of the growth of Lactobacillus plantarum AR514 in a medium with different prebiotics as carbon source in an example of the invention;
FIG. 6 is a graph of the growth of Lactobacillus plantarum AR113 in a simulated mouse gut environment with different prebiotics according to an embodiment of the invention;
FIG. 7 is a graph of the growth of Lactobacillus plantarum AR117 in a simulated mouse gut environment with different prebiotics in an example of the invention;
FIG. 8 is a graph of the growth of Lactobacillus plantarum AR237 in a simulated mouse gut environment in various prebiotics according to an embodiment of the invention;
FIG. 9 is a graph of the growth of Lactobacillus plantarum AR509 in a simulated mouse gut environment with different prebiotics in an example of the invention;
fig. 10 is a graph of the growth of lactobacillus plantarum AR514 in a simulated mouse gut environment with different prebiotics in an example of the invention.
Detailed Description
In order to make the technical means, the creation features, the achievement purposes and the effects of the invention easy to understand, the invention is specifically described below by combining the embodiment and the attached drawings.
Specifically, the sources of the raw materials used in the following examples are as follows:
lactobacillus plantarum AR113 strain has been deposited in China general microbiological culture Collection center (address: no. 3, xilu No.1, beijing, chaoyang, north Chen) in 2017, 03 and 22 months, and its number is CGMCC No.13909.
Lactobacillus plantarum AR237 strain has been deposited in China general microbiological culture Collection center (address: no. 3 Xilu No.1 Beijing, chaoyang, north Chen) in 2017 at 07/04, and its number is CGMCC No.14003.
The Lactobacillus plantarum (Lactobacillus plantarum) AR117 strain, the Lactobacillus plantarum (Lactobacillus plantarum) AR509 strain and the Lactobacillus plantarum (Lactobacillus plantarum) AR514 strain are obtained by self-screening in a laboratory by the inventors.
MRS culture medium formula: 10.0g of peptone, 2.0g of dipotassium phosphate, 10.0g of beef extract powder, 5.0g of yeast extract, 0.25g of manganese sulfate, 5.0g of anhydrous sodium acetate, 20.0g of glucose, 2.0g of diamine citrate, 0.58g of magnesium sulfate, tween-801mL and 1000mL of deionized water.
< example >
Experiments are carried out by taking lactobacillus plantarum AR113, AR117, AR237, AR509 and AR514 as research objects, and the specific steps are as follows:
the specific experimental procedures were as follows:
five strains (AR 113, AR117, AR237, AR509 and AR 514) are taken out from a refrigerator at the temperature of-80 ℃, streaked on an MRS solid medium, cultured and activated in an anaerobic incubator for 48 hours, and preserved by a glycerol preservation method after being repeated three times for standby.
The five activated strains are inoculated into an MRS culture medium according to the inoculation amount of 1 percent, 2 percent of the prebiotics (namely 20ug/ml FOS, 20ug/ml XOS, 20ug/ml GOS, 20ug/ml IMO and 20ug/ml Inulin) are respectively added into each strain culture medium, wherein all the added prebiotics are filtered by a 0.22um water phase filter to keep the added prebiotics in a sterile state, no prebiotics are added into a Control group (Control), and the OD value of the strain at 600nm is measured by using a growth curve instrument.
The results of the experiment are shown in table 1.
TABLE 1 absorbance values of Lactobacillus plantarum in MRS media supplemented with different prebiotics
As shown in Table 1, after different 20. Mu.g/ml prebiotics are added to the MRS culture medium, IMO has obvious promotion effect on each Lactobacillus plantarum in the middle stage of culture. The difference of absorbance of IMO group and the control group is obvious (P < 0.05) when five strains of lactobacillus plantarum are at 24h and 32h, while the proliferation effect of other prebiotic groups on the strains is not obvious, and the growth state of some prebiotic groups is obviously inhibited compared with the control group due to the change of the substrate concentration (P < 0.05), such as FOS group of AR113, XOS group of XOS, inulin group of AR117, XOS group and GOS group of AR237, FOS group of AR509, XOS group, inulin group, XOS group of AR514, GOS group and Inulin group. In the five strains, except for the IMO groups of AR113, AR509 and AR514, the secondary growth phenomenon occurs in the late stage of fermentation culture, and the secondary growth phenomenon does not occur in the IMO groups of the five strains. This is probably because the Lactobacillus plantarum can be simultaneously assimilated and utilized to promote the growth and propagation of the strain when glucose and isomaltooligosaccharide are used as the composite carbon source, and the strain preferentially utilizes glucose as the carbon source after other prebiotics are compounded with glucose, thereby producing catabolite repression effect. Therefore the direct addition of prebiotics in MRS medium cannot be used as a screen for prebiotics suitable for lactobacillus plantarum.
And 2, replacing glucose in the MRS culture medium with the prebiotics in the step 1, culturing the lactobacillus plantarum by using the prebiotics as a carbon source, and determining a growth curve.
The specific experimental steps are as follows:
preparing MRS basal medium without glucose, replacing glucose with 2% of the prebiotics (namely 20ug/ml FOS, 20ug/ml XOS, 20ug/ml GOS, 20ug/ml IMO and 20ug/ml Inulin) as carbon source, inoculating five activated lactobacillus plantarum strains with 1% inoculation amount in the basal medium, measuring absorbance at 600nm by using a growth curve instrument and drawing a growth curve.
The test results are shown in FIGS. 1-5.
FIG. 1 is a graph of the growth of Lactobacillus plantarum AR113 in a medium with different prebiotics as carbon source in an example of the invention. FIG. 2 is a graph showing the growth of Lactobacillus plantarum AR117 in a medium with different prebiotics as carbon source in an example of the present invention. FIG. 3 is a graph showing the growth of Lactobacillus plantarum AR237 in a medium with different prebiotics as carbon source in an example of the present invention. FIG. 4 is a graph showing the growth of Lactobacillus plantarum AR509 in a medium with different prebiotics as carbon source in an example of the present invention. FIG. 5 is a graph showing the growth of Lactobacillus plantarum AR514 in a medium with different prebiotics as carbon source in an example of the present invention.
As shown in FIGS. 1-5, the absorption and utilization of FOS, GOS and IMO by AR113 are good, the growth is very vigorous, the growth is second to Inulin, and the growth is very slow when XOS is used as a carbon source; AR117 has good growth condition in MRS culture medium taking FOS and GOS as carbon sources, and has poor growth condition of IMO, inulin times and XOS; AR237 has good growth condition in MRS culture medium with FOS and Inulin as carbon source, IMO and GOS have inferior and XOS has poor growth condition; AR509 has good growth condition in MRS culture medium taking FOS and GOS as carbon sources, and has poor growth condition of Inulin and XOS after IMO; AR514 grows well in MRS culture medium with Inulin and FOS as carbon source, and GOS and IMO grow less frequently and XOS grows less frequently. 2 kinds of suitable prebiotics are screened out for each lactobacillus plantarum to continue the experiment by integrating the growth conditions of each strain. Therefore, the addition of prebiotics in MRS medium without glucose can be used as a screen for prebiotics suitable for the growth of Lactobacillus plantarum.
And 3, simulating the intestinal environment of the mouse in vitro, performing liquid culture on different lactobacillus plantarum by using the sterilized excrement of the mouse and the prebiotics obtained by screening, and drawing a growth curve, thereby exploring the absorption effect of the lactobacillus plantarum on the prebiotics in the simulated intestinal environment.
The specific experimental steps are as follows: preparing a liquid culture medium of mouse feces: 6ug/ml mouse feces +20ug/ml tryptone. According to the experimental result of the step 2, two kinds of prebiotics with good absorption effect of each strain are selected, the activated bacteria liquid is centrifuged at 9600rpm for 1min, supernatant liquid is poured out, the bacteria liquid is washed by sterile water and then inoculated into a liquid culture medium of the mouse excrement with the inoculation amount of 1%, corresponding prebiotics are respectively added, the concentration is 1%, namely 10ug/ml, and the instrument measures the absorbance at 600nm and then draws the growth curve. The specific grouping of step 3 is shown in table 2.
TABLE 2 Experimental design groupings
The results of the experiments are shown in FIGS. 6-10.
As shown in fig. 6 to 10, when each lactobacillus plantarum was fermented in vitro in a mouse fecal broth medium, the growth conditions of each strain were greatly different from those in MRS medium due to the change in nutrient composition. The growth of AR113 and AR509 was good although the number of bacteria was significantly reduced during the fermentation, and FOS and GOS could be absorbed as carbon sources. AR117 grew slower than AR113 and AR509 and did not adapt well to the simulated mouse intestinal environment. AR237 and AR514 grew well when FOS was used as a carbon source, and were similar to AR113 and AR509, while when Inulin was used as a carbon source, the effect of absorption and utilization was significantly reduced compared to the case of the medium in MRS (FIG. 3 and FIG. 5).
In conclusion, IMO has the effect of promoting the proliferation of each plant lactobacillus in the middle stage of fermentation at the concentration level of 20 ug/ml. While other prebiotics can not play a role in promoting growth of the lactobacillus plantarum at the concentration level of 20ug/ml, and some prebiotics also cause the effect of matrix inhibition. Among five lactobacillus plantarum strains, AR117 has weak adaptability to simulated intestinal environments, and the other four strains have strong adaptability. Of the five prebiotics, FOS and GOS can be absorbed and utilized by the strain in the intestinal microenvironment better, and the growth of the strain is promoted.
Effects and effects of the embodiments
According to the application of the prebiotics in the intestinal environment to promote the growth of lactobacillus plantarum, the provided fructo-oligosaccharides and galacto-oligosaccharides can promote the growth of specific lactobacillus plantarum because the fructo-oligosaccharides and the galacto-oligosaccharides can be absorbed and utilized by specific lactobacillus plantarum strains in the intestinal microenvironment, so that the number of specific lactobacillus plantarum is increased remarkably in a certain period of time.
According to the application of isomaltooligosaccharide in promoting proliferation of lactobacillus plantarum in the middle fermentation stage, when glucose and isomaltooligosaccharide are used as a composite carbon source, lactobacillus plantarum can be absorbed and utilized simultaneously to promote growth and propagation of strains, each strain of lactobacillus plantarum has the proliferation promoting effect in the middle fermentation stage, other prebiotics cannot play a remarkable growth promoting effect on lactobacillus plantarum in the same concentration level, and some prebiotics also cause a matrix inhibition effect.
The above embodiments are preferred examples of the present invention, and are not intended to limit the scope of the present invention.
Claims (1)
1. The application of the prebiotics in promoting the growth of the lactobacillus plantarum in the intestinal environment is characterized in that the prebiotics are fructo-oligosaccharide,
wherein the lactobacillus plantarum is lactobacillus plantarum AR113 or lactobacillus plantarum AR237,
the concentration of the prebiotics is 10 mug/mL-30 mug/mL.
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