CN111004735A - Lactobacillus fermentum and application thereof in improving intestinal health - Google Patents
Lactobacillus fermentum and application thereof in improving intestinal health Download PDFInfo
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Abstract
The invention discloses lactobacillus fermentum and application thereof in improving intestinal health, and belongs to the technical field of microorganisms and medicines. The Lactobacillus fermentum (CCFM 1037) obtained by screening can improve the intestinal health, and is specifically embodied in that: (1) can grow and metabolize to produce acid by taking fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide as unique carbon sources, and has broad-spectrum oligosaccharide utilization capacity; (2) in vitro metabolism can generate a plurality of short chain fatty acids such as acetic acid, propionic acid, butyric acid and the like; (3) the relative abundance of the Parabacteroidides in the intestinal tract can be remarkably reduced, so that the Lactobacillus fermentum CCFM1037 has a huge application prospect in preparing products (such as food, medicines or health care products) for improving the intestinal tract health.
Description
Technical Field
The invention relates to lactobacillus fermentum and application thereof in improving intestinal health, and belongs to the technical field of microorganisms and medicines.
Background
The intestinal micro-ecology is the most important and complex ecological environment of the human body, and the intestinal flora is one of the most active components of the intestinal micro-ecology. The microbial species in the human intestinal tract can reach 500-1000, wherein, the phylum firmicutes, bacteroidetes, actinomycetes, proteobacteria and verrucomicrobia are the main body parts of the normal human intestinal tract flora structure.
The intestinal flora structure is a co-selection of microorganisms and their hosts, which interact to maintain functional stability of the intestinal micro-ecosystem. Generally, if the intestinal flora of a healthy host is in a dynamic equilibrium state, if the equilibrium is disturbed by the influence of diet, environment, and use of antibiotics, the change of intestinal permeability is caused, and then the transfer of bacteria and metabolites thereof is caused, thereby causing gastrointestinal diseases of the host and having the possibility of generating a series of complications. Therefore, maintaining the intestinal flora balance is very important for human health.
The Parabacteroides is a pathogenic bacterium existing in human intestinal tracts, and if the intestinal flora of a human body is unbalanced, the Parabacteroides can be greatly proliferated in the human body, so that diseases such as appendicitis, celiac inflammation, bacteremia and the like of the human body are caused; in addition, because the antibiotics cannot selectively inhibit the Parabacteroides, and the Parabacteroides is easy to acquire drug resistance in the antibiotic application process, in addition, researches show that the low-dose penicillin can even improve the relative abundance of the Parabacteroides in the intestinal tract and increase the risk of bacteremia, so that the infection of the Parabacteroides in a human body is difficult to treat, and a method for inhibiting the Parabacteroides in the intestinal tract is urgently needed to be found.
At present, although some studies have been made on obtaining some probiotics with improved intestinal flora imbalance by screening, for example, patent application publication No. CN107312726A discloses a Lactobacillus plantarum that can inhibit the growth of harmful bacteria such as Escherichia coli, Salmonella, Streptococcus suis and Staphylococcus aureus in intestinal tract; patent application publication No. CN104232515A discloses Bifidobacterium animalis which is capable of increasing the number of bifidobacteria and lactobacilli in the intestinal tract and, at the same time, of decreasing the number of bacteroides and coliform bacteria in the intestinal tract.
However, the probiotics lack pertinence to the Parabacteroides, and at present, no probiotics capable of remarkably inhibiting the Parabacteroides in human intestinal tracts exists, so that the treatment of the Parabacteroides infection is undoubtedly prevented.
Disclosure of Invention
[ problem ] to
The invention aims to solve the technical problem of providing a Lactobacillus fermentum which can obviously inhibit the relative abundance of the Parabacteroides in human intestinal tracts; the relative abundance is the percentage of the number of a particular species of bacteria in a sample relative to the total number of bacteria in the sample.
[ solution ]
In order to solve the problems, the invention provides a Lactobacillus fermentum (CCFM 1037), wherein the Lactobacillus fermentum (CCFM 1037) is stored in Guangdong province microorganism strain preservation center in 2018, 11 and 20 days, and the preservation number is GDMCC No.60488, and the preservation address is No. 59 building 5 of Michelia furiosaefolia Miyao No. 100, Guangzhou city.
The Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037 is obtained by separating village excrement samples from old town relay village in Chengmei county of Hainan province, sequencing analysis is carried out on the strain, the 16S rRNA sequence of the strain is shown as SEQ ID NO.1, and the sequence obtained by sequencing is compared with the nucleic acid sequence in NCBI, so that the strain is Lactobacillus fermentum and is named as Lactobacillus fermentum CCFM 1037.
The Lactobacillus fermentum (CCFM 1037) has the following characteristics:
(1) the bacterial colony cultured for 48 hours on the MRS culture medium is round, white, transparent and smooth;
(2) resistance to simulated gastrointestinal fluids: after 3 hours of culture in 3g/L of pepsin-containing physiological saline with pH of 3, the survival rate is as high as 110.9 +/-7.58%; after 4h of culture in physiological saline at pH 8 containing 1g/L trypsin and 0.3% (m/v) (i.e., 0.3g/100mL) bile salts, the survival rate was 14.15 + -5.54%; after the culture is carried out for 3 hours in physiological saline with pH 3 and 3g/L pepsin, the culture is continued for 4 hours in physiological saline with pH 8 and containing 1g/L trypsin and 0.3% (m/v) bile salt, and the survival rate is still 15.69 +/-0.42%;
(3) can grow and metabolize to produce acid by taking fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide as unique carbon sources, and has broad-spectrum oligosaccharide utilization capacity;
(4) in vitro metabolism can generate a plurality of short chain fatty acids such as acetic acid, propionic acid, butyric acid and the like;
(5) can obviously reduce the relative abundance of the Parabacteroides in the intestinal tract.
The invention provides application of the Lactobacillus fermentum (CCFM 1037) in preparing a product for improving intestinal health.
In one embodiment of the invention, the viable count of Lactobacillus fermentum (CCFM 1037) in the product is not less than 1 × 106CFU/mL or 1X 106CFU/g。
In one embodiment of the invention, the viable count of Lactobacillus fermentum (CCFM 1037) in the product is not less than 1 × 108CFU/mL or 1X 108CFU/g。
In one embodiment of the invention, the product comprises a food, pharmaceutical or nutraceutical product.
In one embodiment of the invention, the medicament comprises lactobacillus fermentum (CCFM 1037), a pharmaceutical carrier and/or a pharmaceutical excipient.
The invention provides a product for improving intestinal health, which contains the Lactobacillus fermentum (CCFM 1037).
In one embodiment of the invention, the viable count of Lactobacillus fermentum (CCFM 1037) in the product is not less than 1 × 106CFU/mL or 1X 106CFU/g。
In one embodiment of the invention, the viable count of Lactobacillus fermentum (CCFM 1037) in the product is not less than 1 × 108CFU/mL or 1X 108CFU/g。
In one embodiment of the invention, the product comprises a food, pharmaceutical or nutraceutical product.
In one embodiment of the invention, the medicament comprises lactobacillus fermentum (CCFM 1037), a pharmaceutical carrier and/or a pharmaceutical excipient.
Has the advantages that:
the invention screens out a Lactobacillus fermentum (CCFM 1037), the Lactobacillus fermentum (CCFM1037 can improve the intestinal health, and the specific expression is as follows:
(1) functional oligosaccharide can not be digested and absorbed by stomach and small intestine, but can be utilized by probiotics in large intestine, beneficial bacteria can be proliferated, harmful bacteria can be inhibited, and intestinal flora balance can be adjusted, while lactobacillus fermentum (CCFM 1037) can grow and metabolize acid by taking fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide as unique carbon sources, and has broad-spectrum oligosaccharide utilization capability;
(2) short-chain fatty acids can reduce the pH value in intestinal tracts, improve the acidic environment of the intestinal tracts, proliferate beneficial bacteria, inhibit harmful bacteria to adjust intestinal flora and improve the intestinal functions, while the in vitro metabolism of the Lactobacillus fermentum (CCFM 1037) can generate a plurality of short-chain fatty acids such as acetic acid, propionic acid and butyric acid, and the content of the short-chain fatty acids in the mouse excrement is obviously increased after the Lactobacillus fermentum (CCFM 103715 days);
(3) the Parabacteroides belongs to a harmful bacterium which is easy to generate resistance to antibiotics and exists in human intestinal tracts, and can cause diseases such as appendicitis, abdominal inflammation, bacteremia and the like of organisms, the Lactobacillus (Lactobacillus fermentum) CCFM1037 can obviously reduce the relative abundance of the Parabacteroides in the intestinal tracts, and after the Lactobacillus (Lactobacillus fermentum) CCFM103715 days, the relative abundance of the Parabacteroides in mouse excrement is obviously reduced,
therefore, the Lactobacillus fermentum (CCFM 1037) has a huge application prospect in preparing products (such as food, medicines or health care products) for improving intestinal health.
Biological material preservation
A Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037 is classified and named as Lactobacillus fermentum, and is preserved in Guangdong province microorganism strain preservation center in 11 and 20 months in 2018, wherein the preservation number is GDMCC No.60488, and the preservation address is No. 59 building 5 of Dazhou No. 100 of Xielianlu Guangzhou city.
Drawings
FIG. 1: culturing different lactobacillus fermentum in simulated gastric fluid for 3h, and culturing in simulated intestinal fluid for 4 h; wherein the abscissa is the number of Lactobacillus fermentum (LF 1, LF2, LF3, LF4, LF5, LF6, LF7, LF8, LF9, LF10, CCFM 1037.
FIG. 2: the yield of total short-chain fatty acids of different lactobacillus fermentum strains in the in vitro culture process; wherein the abscissa is the number of Lactobacillus fermentum (LF 1, LF2, LF3, LF4, LF5, LF6, LF7, LF8, LF9, LF10, CCFM 1037.
FIG. 3: the yield of acetic acid of different strains of lactobacillus fermentum in the in vitro culture process; wherein the abscissa is the number of Lactobacillus fermentum (LF 1, LF2, LF3, LF4, LF5, LF6, LF7, LF8, LF9, LF10, CCFM 1037.
FIG. 4: the yield of propionic acid of different lactobacillus fermentum strains in the in vitro culture process; wherein the abscissa is the number of Lactobacillus fermentum (LF 1, LF2, LF3, LF4, LF5, LF6, LF7, LF8, LF9, LF10, CCFM 1037.
FIG. 5: the yield of butyric acid of different lactobacillus fermentum strains in the in vitro culture process; wherein the abscissa is the number of Lactobacillus fermentum (LF 1, LF2, LF3, LF4, LF5, LF6, LF7, LF8, LF9, LF10, CCFM 1037.
FIG. 6: relative abundance of parabacteroidides in feces of CCFM1037 group of mice in example 9, where "×" indicates a significant difference from the blank group (P < 0.05).
Detailed Description
The invention is further illustrated with reference to specific examples.
Pepsin 1:10000U (product No. A600688, CAS: [9001-75-6]) referred to in the following examples was purchased from Biotechnology engineering (Shanghai) Ltd; trypsin 1:250 (product No. 64008867, CAS: [9002-07-7]) referred to in the following examples was purchased from national pharmaceutical group chemical Co., Ltd; bile salts referred to in the following examples were purchased from Shanghai Bayer Biotech Ltd; glucose referred to in the following examples was purchased from national pharmaceutical group chemical agents limited; fructooligosaccharides (FOS) and Galactooligosaccharides (GOS) referred to in the following examples were purchased from cheng bao bio-inc; xylo-oligosaccharides (XOS) referred to in the following examples were purchased from shanghai-sourced leaf biotechnology limited; the skim milk powders referred to in the examples below were purchased from illite.
The media involved in the following examples are as follows:
MRS liquid medium (g/L): 10g/L of peptone, 5g/L of yeast extract, 20g/L of glucose, 2g/L of anhydrous sodium acetate and 2g/L, K of citric acid hydrogen diamine2HPO4·3H2O 2.6g/L、MgSO4·7H2O 0.5g/L、MnSO4·7H2O0.25 g/L, Tween-801 g/L and distilled water 1000 g/L.
MRS solid medium (g/L): 10g/L of peptone, 5g/L of yeast extract, 20g/L of glucose, 2g/L of anhydrous sodium acetate and hydrogen citrateDiamine 2g/L, K2HPO4·3H2O 2.6g/L、MgSO4·7H2O 0.5g/L、MnSO4·7H2O0.25 g/L, Tween-801 g/L, agar 20g/L and distilled water 1000 g/L.
Example 1: screening and strain identification of lactobacillus fermentum
1. Screening
Taking village feces from old town Luo Ying village of Chengmei county of Hainan province as sample, and performing gradient dilution with sterile physiological saline 10 times to 10 times-6Then 100. mu.L of each dilution gradient was set to 10-4、10-5、10-6The diluted solution is plated on an MRS solid culture medium, cultured for 48 hours at 37 ℃, and observed and recorded colony morphology; selecting colonies with different forms on an MRS solid culture medium for streaking separation, and after culturing for 48 hours at 37 ℃, selecting single colonies with different forms on the MRS solid culture medium again for streaking separation until obtaining pure single colonies with consistent forms; selecting pure colonies on an MRS solid culture medium, inoculating the pure colonies in a 5mLMRS liquid culture medium, and culturing for 18h at 37 ℃; and (3) taking 1mL of bacterial liquid in a sterile centrifuge tube, centrifuging for 3min at 8000r/min, removing an upper culture medium, resuspending bacterial sludge in a 30% glycerol solution, and preserving at-80 ℃ to obtain the bacterial strain.
2. Identification
The isolated strains were subjected to PCR amplification of 16S rDNA, the PCR products were sent to Chiba Gene sequencing Co., Ltd for sequencing (the 16S rDNA sequence of CCFM1037 is shown in SEQ ID NO. 1), and the results of sequencing were compared with the nucleic acid sequence in NCBI to finally obtain 11 strains of fermented milk bacteria, which were respectively named Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037, Lactobacillus fermentum (Lactobacillus fermentum) LF1, Lactobacillus fermentum (Lactobacillus fermentum) LF2, Lactobacillus fermentum (Lactobacillus fermentum) LF3, Lactobacillus fermentum (Lactobacillus fermentum) LF4, Lactobacillus fermentum (Lactobacillus fermentum) 5, Lactobacillus (Lactobacillus fermentum) LF6, Lactobacillus fermentum (Lactobacillus) 5639 LF7, Lactobacillus fermentum (Lactobacillus) 8, Lactobacillus fermentum (Lactobacillus fermentum) 9, and Lactobacillus fermentum (Lactobacillus fermentum) LF 10.
Example 2: culture of Lactobacillus fermentum
After Lactobacillus fermentum (CCFM 1037) is inoculated to MRS solid culture medium and cultured for 48h at 37 ℃, the colony is observed and is found to be round, white, transparent and smooth.
Example 3: tolerance of different lactobacillus fermentum to simulated gastrointestinal fluids
The method comprises the following specific steps:
1. tolerance of different lactobacillus fermentum to simulated gastric juice
The Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037, the Lactobacillus fermentum (Lactobacillus fermentum) LF1, the Lactobacillus fermentum (Lactobacillus fermentum) LF2, the Lactobacillus fermentum (Lactobacillus fermentum) LF3, the Lactobacillus fermentum) LF4, the Lactobacillus fermentum (Lactobacillus fermentum) LF5, the Lactobacillus fermentum (Lactobacillus fermentum) LF6, the Lactobacillus fermentum (Lactobacillus fermentum) LF7, the Lactobacillus fermentum (Lactobacillus fermentum) LF8, the Lactobacillus fermentum) LF9, and the Lactobacillus fermentum (Lactobacillus fermentum) LF10 were respectively inoculated into an MRS liquid medium for 18h at 37 ℃, cells were collected by centrifugation in physiological saline, washed, and then collected again in pH adjusted by centrifugation in physiological saline, pH 3/pH 3 of the liquid containing the pepsin LF3600And 5.0, taking 1mL of bacterial liquid to perform flat viable count to serve as the original viable count of the lactobacillus fermentum in the bacterial liquid, placing the residual bacterial liquid at 37 ℃ for culturing for 3 hours, and taking 1mL of bacterial liquid to perform flat viable count to serve as the viable count of the lactobacillus fermentum in the bacterial liquid after tolerating simulated gastric juice.
Wherein the survival rate (%) after the gastric juice tolerance (viable count of lactobacillus fermentum in the bacterial liquid after the simulated gastric juice tolerance/original viable count of lactobacillus fermentum in the bacterial liquid) is 100%.
The detection results are as follows: after 3h of cultivation in physiological saline containing pepsin at pH 3, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037 was 110.46 + -5.66%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF1 was 97.08 + -2.15%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF2 was 110.84 + -4.98%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF3 was 97.62 + -1.46%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF4 was 114.89 + -16.49%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF5 was 192.95 + -0.82%, the survival rate of Lactobacillus (Lactobacillus fermentum) LF 38 was 48 + -8.97%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF7 was 585 + -8.97%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF 638.89%, the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF 6326 was 585.64%, and the survival rate of Lactobacillus fermentum (Lactobacillus fermentum) LF 89% was 89%.
It can be seen that Lactobacillus fermentum (CCFM1037, LF2, LF4, LF5, LF6, LF7, LF8 are more resistant to simulated gastric juice.
2. Tolerance of different lactobacillus fermentum to simulated intestinal fluid
The Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037, the Lactobacillus fermentum (Lactobacillus fermentum) LF1, the Lactobacillus fermentum (Lactobacillus fermentum) LF2, the Lactobacillus fermentum (Lactobacillus fermentum) LF3, the Lactobacillus fermentum) LF4, the Lactobacillus fermentum (Lactobacillus fermentum) LF5, the Lactobacillus fermentum (Lactobacillus fermentum) LF6, the Lactobacillus fermentum (Lactobacillus fermentum) LF7, the Lactobacillus fermentum (Lactobacillus fermentum) LF8, the Lactobacillus fermentum) LF9, and the Lactobacillus fermentum (Lactobacillus fermentum) LF10 were respectively inoculated into an MRS liquid medium for 18h at 37 ℃, the cells were collected by centrifugation in physiological saline, washed, and then collected again in pH adjusted by pH 0/pH 36005.0, taking 1mL of the bacterial liquid to perform flat viable count to serve as the original viable count of the lactobacillus fermentum in the bacterial liquid, putting the rest of the bacterial liquid at 37 ℃ for culturing for 4h, taking 1mL of the bacterial liquid to perform flat viable count to serve as the tolerant simulated intestinal liquid, and then fermenting the bacterial liquidViable count of lactobacillus.
Wherein the survival rate (%) after the intestinal fluid tolerance (viable count of lactobacillus fermentum in the bacterial liquid after the intestinal fluid simulation tolerance/original viable count of lactobacillus fermentum in the bacterial liquid) is 100%.
The detection results are as follows: after 4h of cultivation in physiological saline containing trypsin and bile salts at a pH of 8, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) CCFM1037 was 14.20 + -0.35%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF1 was 4.89 + -0.65%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF2 was 1.56 + -0.02%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF3 was 1.11 + -0.03%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF4 was 0.40 + -0.05%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) 5 was 1.56 + -0.03%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF 78 was 8.12 + -0.04%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF 7.56 + -0.03%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) LF6 was 8.12 + -0.04%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) was 9%, the survival rate of the fermented milk bacteria (Lactobacillus fermentum) was 8%, and the survival rate of the fermented milk bacteria (Lactobacillus fermentum) was 9%.
It can be seen that Lactobacillus fermentum (CCFM1037, LF1, and LF6 have strong tolerance to simulated intestinal fluid.
3. Tolerance of different lactobacillus fermentum to simulated gastrointestinal fluids
The fermented milk bacillus (Lactobacillus fermentum) CCFM1037, the fermented milk bacillus (Lactobacillus fermentum) LF1, the fermented milk bacillus (Lactobacillus fermentum) LF2, the fermented milk bacillus (Lactobacillus fermentum) LF3, the fermented milk bacillus (Lactobacillus fermentum) LF4, the fermented milk bacillus (Lactobacillus fermentum) LF5, the fermented milk bacillus (Lactobacillus fermentum) LF6, the fermented milk bacillus (Lactobacillus fermentum) LF7, the fermented milk bacillus (Lactobacillus fermentum) LF8, the fermented milk bacillus (Lactobacillus fermentum) LF9 and the fermented milk bacillus (Lactobacillus fermentum) LF10 obtained in example 1 were respectively inoculated into an MRS liquid medium and cultured at 37 ℃ for 18 hours, and then the cells were collected by centrifugation, and the cultured cells were collectedWashing the collected cells with normal saline, centrifuging again to collect the cells after washing, and respectively suspending the collected cells in normal saline with pH of 3(pH adjusted by HCl) and containing 3g/L pepsin until the cells reach bacterial liquid OD6005.0, taking 1mL of bacterial liquid to perform plate viable count to obtain the original viable count of lactobacillus fermentum in the bacterial liquid, culturing the rest of bacterial liquid at 37 ℃ for 3h, centrifuging the cultured bacterial liquid to collect cells, and resuspending the collected cells in physiological saline with pH 8(pH adjusted by NaOH) containing 1g/L trypsin and 0.3% (m/v) cholate to obtain bacterial liquid OD6005.0, after culturing at 37 ℃ for 4h, 1mL of the broth was taken for viable flat count as the viable count of Lactobacillus fermentum in the broth after tolerating simulated gastrointestinal fluids.
Wherein the survival rate after gastrointestinal fluid tolerance (%) (% viable count of lactobacillus fermentum in a bacterial liquid after gastrointestinal fluid tolerance/original viable count of lactobacillus fermentum in the bacterial liquid) is 100%.
The detection results are as follows: as is clear from FIG. 1, after culturing for 3 hours in physiological saline containing pepsin at pH 3, the survival rates of fermented milk bacteria (Lactobacillus fermentum) CCFM1037 and Lactobacillus fermentum (Lactobacillus fermentum) LF1 and Lactobacillus fermentum (Lactobacillus fermentum) LF2 were 15.69 + -0.42%, 1.08 + -0.02%, 0.46 + -0.00%, 3.3902%, 3.43 + -0.04%, 3.43 + -0.69%, and 3.69%, respectively, 4.69%, and 80%, respectively, after culturing for 3 hours in physiological saline containing pepsin at pH 8, 4.05%, and 80%, respectively, the survival rates of fermented milk bacteria (Lactobacillus fermentum) LF 598, 4.73%, 3.06%, and 3.69%, respectively, The survival rate of Lactobacillus fermentum LF10 was 0.39. + -. 0.01%.
It can be seen that Lactobacillus fermentum (CCFM1037, LF6, LF7 are more tolerant to simulated gastrointestinal fluids.
Example 4: oligosaccharide utilization capacity of different lactobacillus fermentum
The method comprises the following specific steps:
(1) on the basis of MRS solid culture medium, removing glucose and beef extract in the formula, taking no sugar as a blank control, respectively adding 0.5% (m/v) of glucose, fructo-oligosaccharide, xylo-oligosaccharide and galacto-oligosaccharide as carbon sources, and adding bromocresol purple as an acid-base indicator to obtain a solid culture medium plate which is sugar-free, contains glucose, fructo-oligosaccharide, xylo-oligosaccharide or galacto-oligosaccharide;
(2) the Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037, the Lactobacillus fermentum (Lactobacillus fermentum) LF1, the Lactobacillus fermentum (Lactobacillus fermentum) LF2, the Lactobacillus fermentum (Lactobacillus fermentum) LF3, the Lactobacillus fermentum (Lactobacillus fermentum) LF4, the Lactobacillus fermentum (Lactobacillus fermentum) LF5, the Lactobacillus fermentum (Lactobacillus fermentum) LF6, the Lactobacillus fermentum (Lactobacillus fermentum) LF7, the Lactobacillus fermentum) LF8, the Lactobacillus fermentum (Lactobacillus fermentum) LF9, and the Lactobacillus fermentum (Lactobacillus fermentum) LF10 were respectively inoculated into an MRS liquid medium and cultured at 37 ℃ for 18h, and after centrifugation in physiological saline, the cells were respectively resuspended in physiological saline to OD600Obtaining bacterial liquid when the concentration is 0.5;
(3) and (3) sucking 10 mu L of bacterial liquid, respectively dropping the bacterial liquid on the solid culture medium plate obtained in the step (1), after the bacterial liquid is completely absorbed, carrying out inverted culture at 37 ℃, after 12h, observing whether a bromocresol purple indicator in the solid culture medium plate turns yellow or not, wherein if the bromocresol purple indicator turns yellow, the carbon source is utilized, and if the bromocresol purple indicator does not turn yellow, the carbon source is not utilized (the result is shown in table 1).
As can be seen from table 1, Lactobacillus fermentum generally has a good ability to utilize oligosaccharides, and can widely utilize fructo-oligosaccharides, xylo-oligosaccharides, and galacto-oligosaccharides to produce acids, so as to yellow the bromocresol purple indicator in the solid medium.
TABLE 1 ability of different L.fermentum to utilize different carbon sources
| Group of | Sugar-free | Glucose | Fructo-oligosaccharide | Xylo-oligosaccharide | Galacto-oligosaccharides |
| CCFM1037 | - | + | + | + | + |
| LF1 | - | + | + | + | + |
| LF2 | - | + | + | + | + |
| LF3 | - | + | + | + | + |
| LF4 | - | + | + | + | + |
| LF5 | - | + | + | + | + |
| LF6 | - | + | + | + | + |
| LF7 | - | + | + | + | + |
| LF8 | - | + | + | + | + |
| LF9 | - | + | + | + | + |
| LF10 | - | + | + | + | + |
Wherein, + indicates that the Lactobacillus can utilize the carbon source, -indicates that it cannot.
Example 5: short chain fatty acid producing ability of different lactobacillus fermentum
The method comprises the following specific steps:
the Lactobacillus fermentum (Lactobacillus fermentum) CCFM1037, the Lactobacillus fermentum (Lactobacillus fermentum) LF1, the Lactobacillus fermentum (Lactobacillus fermentum) LF2, the Lactobacillus fermentum (Lactobacillus fermentum) LF3, the Lactobacillus fermentum (Lactobacillus fermentum) LF4, the Lactobacillus fermentum (Lactobacillus fermentum) LF5, the Lactobacillus fermentum (Lactobacillus fermentum) LF6, the Lactobacillus fermentum (Lactobacillus fermentum) LF7, the Lactobacillus fermentum) LF8, the Lactobacillus fermentum (Lactobacillus fermentum) LF9, and the Lactobacillus (Lactobacillus fermentum) LF10 were respectively inoculated into an MRS liquid medium and cultured at 37 ℃ for 18h, and after centrifugation in physiological saline, the cells were respectively resuspended in physiological saline to a concentration of 2 × 108CFU/mL, obtaining bacterial liquid.
Mixing and acidifying the bacterial liquid and 10% sulfuric acid at a volume ratio of 25:1, adding diethyl ether with the volume 4 times that of the 10% sulfuric acid, oscillating and mixing uniformly to extract fatty acid to obtain an extracting solution; centrifuging the extractive solution at 18000g for 15minSeparating the upper diethyl ether phase, and passing the upper diethyl ether phase over anhydrous Na2SO4Drying, standing for 30min, centrifuging at 18000g for 5min, and analyzing each short chain fatty acid in the upper ether phase by GC-MS (see FIGS. 2-5).
As shown in FIGS. 2 to 5, the total short-chain fatty acid yield of the Lactobacillus fermentum CCFM1037 is 3.641 +/-0.25 mu mol/mL, wherein the acetic acid yield is the highest and is 3.601 +/-0.23 mu mol/mL, which accounts for 98.98% of the total short-chain fatty acid yield and is obviously superior to that of the rest Lactobacillus fermentum.
From examples 3 to 5, it is clear that Lactobacillus fermentum (CCFM 1037), Lactobacillus fermentum (LF 6) and Lactobacillus fermentum (LF 7 are all better able to tolerate simulated gastrointestinal fluids, oligosaccharides utilization and short chain fatty acid production in vitro and able to survive and function in the human gastrointestinal environment, and therefore, Lactobacillus fermentum (CCFM 1037), Lactobacillus fermentum (LF 6) and Lactobacillus fermentum (LF 7) were selected for further experiments to study their probiotic function. .
Example 8: influence of different lactobacillus fermentum on mouse health index, fecal water content and concentration of short-chain fatty acids in feces
The method comprises the following specific steps:
the Lactobacillus fermentum (CCFM 1037), the Lactobacillus fermentum (LF 6) and the Lactobacillus fermentum (LF 7) obtained in example 1 were respectively inoculated into MRS broth and cultured at 37 ℃ for 18 hours, then centrifuged to collect cells, washed with physiological saline, and then respectively resuspended in 12% (m/v) skim milk powder solution to a bacteria concentration of 5 × 109CFU/mL to obtain bacterial suspension, and storing the bacterial suspension at-80 ℃ for later use.
Taking 40 SPF-grade BALB/c male mice with the weight of 20-22 g, randomly dividing the mice into 4 groups, wherein each group comprises 10 mice, and the mice respectively comprise: the kit comprises a blank group, a CCFM1037 group of lactobacillus gasseri CCFM1037 bacterial suspension, an LF6 group of lactobacillus gasseri LF6 bacterial suspension and an LF7 group of lactobacillus gasseri LF7 bacterial suspension, wherein the CCFM1037 group, the LF6 group and the LF7 group are experimental groups.
The experiment took 22 days: the first week (7 days) is the mouse adaptation period; starting intragastric administration on day 8 until the experiment is finished, performing intragastric administration on lactobacillus fermentum CCFM1037, LF6 and LF7 bacterial suspensions respectively at a dose of 0.2mL bacterial suspension/mouse/day, and performing intragastric administration on a blank group by using an equivalent skim milk powder solution as a control; gavage for 15 days.
After 15 days, the fecal content of short-chain fatty acids was measured in each mouse (see Table 2 for the results, and example 5 for the method of measuring the short-chain fatty acid content).
As can be seen from Table 2, the total short-chain fatty acid content in the feces of the mice of the L.fermentum CCFM1037 is 61.63 +/-17.54 mu mol/g, wherein the propionic acid content is 38.99 +/-11.93 mu mol/g, the propionic acid content is 14.12 +/-3.85 mu mol/g, the isobutyric acid content is 0.56 +/-0.18 mu mol/g, the n-butyric acid content is 7.93 +/-3.90 mu mol/g, and the total acid content, the acetic acid content and the n-butyric acid content are all obviously higher than those of the blank group; and various short-chain fatty acids in the mouse feces of the lactobacillus gasseri LF6 and LF7 are higher than those in the blank group, but have no significant difference.
Therefore, the lactobacillus fermentum CCFM1037 can obviously improve the content of short-chain fatty acid in excrement and has obvious advantages.
TABLE 2 types and contents of short-chain fatty acids in feces of mice of different groups
Example 9: effect of different Lactobacillus fermentum on fecal flora in mice
The method comprises the following specific steps:
the Lactobacillus fermentum (CCFM 1037), the Lactobacillus fermentum (LF 6) and the Lactobacillus fermentum (LF 7) obtained in example 1 were respectively inoculated into MRS broth and cultured at 37 ℃ for 18 hours, then centrifuged to collect cells, washed with physiological saline, and then respectively resuspended in 12% (m/v) skim milk powder solution to a bacteria concentration of 5 × 109CFU/mL to obtain bacterial suspension, and culturingThe suspension was stored at-80 ℃ until use.
Taking 40 SPF-grade BALB/c male mice with the weight of 20-22 g, randomly dividing the mice into 4 groups, wherein each group comprises 10 mice, and the mice respectively comprise: the kit comprises a blank group, a CCFM1037 group of lactobacillus gasseri CCFM1037 bacterial suspension, an LF6 group of lactobacillus gasseri LF6 bacterial suspension and an LF7 group of lactobacillus gasseri LF7 bacterial suspension, wherein the CCFM1037 group, the LF6 group and the LF7 group are experimental groups.
The experiment took 22 days: the first week (7 days) is the mouse adaptation period; starting intragastric administration on day 8 until the experiment is finished, performing intragastric administration on lactobacillus fermentum CCFM1037, LF6 and LF7 bacterial suspensions respectively at a dose of 0.2mL bacterial suspension/mouse/day, and performing intragastric administration on a blank group by using an equivalent skim milk powder solution as a control; gavage for 15 days.
Collecting feces of each mouse in the morning of 15 days after gavage, extracting bacterial genome in a feces sample of the mouse by using a feces sample genome extraction Kit purchased from American MP company, amplifying a V4 region of 16S rDNA by using the bacterial genome as a template, performing gel cutting recovery after PCR verification, and quantifying (ng/mu L) the recovered product according to the instruction of a QuantTMdsDNAssay Kit purchased from American Life Technologies company; according to the quantitative result of the PicoGreen fluorescent dye, samples are mixed according to equal mass concentration, and the barcode between the samples is not repeated; constructing a library according to the instruction of TurSeq DNALT Sample Preparation Kit purchased from Life Technologies, USA, and mainly comprising the steps of end repair, 3' end adding A, linker connection, PCR amplification and the like; performing on-machine sequencing on the library; and (3) carrying out offline treatment according to the sequencing result to obtain the distribution condition of the flora in the mouse feces of different experimental groups on the phylum level, the distribution condition of the flora in the mouse feces of different experimental groups on the genus level and the relative abundance of Acinetobacter in the mouse feces of different experimental groups (the detection result is shown in figure 6).
As can be seen from FIG. 6, the relative abundance of Parabacteroides in the feces of mice in the blank group is high and is about 1.70 +/-0.35%, the relative abundance of Parabacteroides in the feces of mice in the small education blank group and Lactobacillus gasseri CCFM1037, LF6 and LF7 is remarkably reduced, wherein the CCFM1037 has the most remarkable reduction amplitude which is only 0.018 +/-0.02%, and the effect of reducing the host infection of the Parabacteroides is the most remarkable.
Therefore, the lactobacillus fermentum CCFM1037 can obviously show the relative abundance of the Parabacteroides in the intestinal tract of the mouse, and has obvious advantages.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Sequence listing
<110> university of south of the Yangtze river
<120> lactobacillus fermentum and application thereof in improving intestinal health
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<170>PatentIn version 3.3
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113151070A (en) * | 2021-04-02 | 2021-07-23 | 江南大学 | Lactobacillus fermentum capable of improving relative abundance of Guttiferae in intestinal tract |
| CN114686393A (en) * | 2020-12-31 | 2022-07-01 | 杭州远大生物制药有限公司 | Bifidobacterium animalis subsp lactis and composition thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012142605A1 (en) * | 2011-04-15 | 2012-10-18 | Samaritan Health Services | Rapid recolonization deployment agent |
| CN107075563A (en) * | 2014-09-30 | 2017-08-18 | 深圳华大基因科技有限公司 | Biomarkers for Coronary Artery Disease |
| CN108384735A (en) * | 2018-03-15 | 2018-08-10 | 江南大学 | Lactobacillus plantarum CCFM1019, its fermented food and its application in medicine preparation |
| CN108495637A (en) * | 2016-01-26 | 2018-09-04 | 雀巢产品技术援助有限公司 | Composition with specific oligosaccharide is by increasing colon SCFA yield and/or by increasing GLP-1 secretions for preventing fat or relevant comorbidities in the future |
-
2019
- 2019-03-21 CN CN201910217288.0A patent/CN111004735A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012142605A1 (en) * | 2011-04-15 | 2012-10-18 | Samaritan Health Services | Rapid recolonization deployment agent |
| CN107075563A (en) * | 2014-09-30 | 2017-08-18 | 深圳华大基因科技有限公司 | Biomarkers for Coronary Artery Disease |
| CN108495637A (en) * | 2016-01-26 | 2018-09-04 | 雀巢产品技术援助有限公司 | Composition with specific oligosaccharide is by increasing colon SCFA yield and/or by increasing GLP-1 secretions for preventing fat or relevant comorbidities in the future |
| CN108384735A (en) * | 2018-03-15 | 2018-08-10 | 江南大学 | Lactobacillus plantarum CCFM1019, its fermented food and its application in medicine preparation |
Non-Patent Citations (3)
| Title |
|---|
| 吉梦馨: ""具有调节肠道动力作用益生乳杆菌的筛选与评价"", 《中国优秀硕士学位论文全文数据库（电子期刊）基础科学辑》 * |
| 沈旭丹等: ""具有肠道动力及肠道菌群调节功能乳杆菌的筛选及功能评价"", 《食品科学》 * |
| 郭蕾: ""乳酸菌对小鼠肠道菌群失衡的调节作用研究"", 《中国优秀硕士学位论文全文数据库（电子期刊）医药卫生科技辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114686393A (en) * | 2020-12-31 | 2022-07-01 | 杭州远大生物制药有限公司 | Bifidobacterium animalis subsp lactis and composition thereof |
| CN114686393B (en) * | 2020-12-31 | 2023-09-05 | 杭州远大生物制药有限公司 | Bifidobacterium animalis subspecies lactis and composition thereof |
| CN113151070A (en) * | 2021-04-02 | 2021-07-23 | 江南大学 | Lactobacillus fermentum capable of improving relative abundance of Guttiferae in intestinal tract |
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