CN107099491A - Lactobacillus plantarum YS3 and its application in the food of Constipation is prepared - Google Patents

Lactobacillus plantarum YS3 and its application in the food of Constipation is prepared Download PDF

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CN107099491A
CN107099491A CN201710542452.6A CN201710542452A CN107099491A CN 107099491 A CN107099491 A CN 107099491A CN 201710542452 A CN201710542452 A CN 201710542452A CN 107099491 A CN107099491 A CN 107099491A
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lactobacillus plantarum
constipation
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骞宇
赵欣
易若琨
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Chongqing University of Education
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    • C12R2001/225Lactobacillus
    • C12R2001/25Lactobacillus plantarum

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Abstract

It is CCTCC NO the invention discloses deposit number:M2016749 Lactobacillus plantarum YS3 (Lactobacillus plantarum YS3) and its application in the food of Constipation is prepared, not only expand Lactobacillus plantarum YS3 application, its application value is improved, and new hope is brought to the prevention of constipation.

Description

Lactobacillus plantarum YS3 and its application in the food of Constipation is prepared
Technical field
The invention belongs to microbial technology field, it is related to a kind of Lactobacillus plantarum and its application in food is prepared.
Background technology
Yak yogurt is a kind of spontaneous fermentation dairy products in Qinghai-Tibet Tibetan area, contains abundant nutritional ingredient, tool There are anti-oxidant, reduction cholesterol, regulation immunity.Due to special spontaneous fermentation condition, including fermentation raw material breast, hair Ferment temperature, fermentation time, fermentation vessel, particularly special fermentative microorganism composition, cause yak yogurt to have flavour And quality.
Constipation state can be considered at when defecation frequency is less than 3 times weekly, when number of times is considered as seriously less than 1 time Constipation state, be it is a kind of threaten health, particularly there is the complicated symptom of considerable influence to colon health.Constipation is usual Disease is not considered as, majority of case can pass through diet and the regulation Constipation of custom.
The content of the invention
Carried out it is an object of the invention to carry out separation identification to the microorganism in yak yogurt, and to its activity and application Research.
Through research, the present invention provides following technical scheme:
1. Lactobacillus plantarum YS3 (Lactobacillus plantarum YS3), deposit number is CCTCC NO: M2016749。
2. applications of the Lactobacillus plantarum YS3 in the food of Constipation is prepared.
The present invention has carried out separation identification to the microorganism in the yak yogurt of Yushu district, Qinghai Tibetan autonomous prefecture, will wherein one Lactobacillus plantarum (Lactobacillus plantarum) is named as YS3, and Chinese Typical Representative is preserved on December 14th, 2016 Culture collection (abbreviation CCTCC, address:No. 299 Wuhan Universitys of Wuhan City, Hubei Province Wuchang District Bayi Road), deposit number For CCTCC NO:M2016749.
Activated carbon inducing constipation model mice result of study shows that Lactobacillus plantarum YS3 anti-hydrochloric acid in gastric juice and cholate ability is strong In lactobacillus bulgaricus, the mouse weight decline that constipation is caused can be suppressed, stool weight, granule number and water content decline, Propulsion rate of the activated carbon in small intestine can be improved simultaneously, reduce the time of the first grain melena of discharge, moreover it is possible to make the blood of mice with constipation Clearing stomach therbligs (MTL), gastrin (Gas), Endothelin (ET), acetylcholinesterase (AChE), Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2 (SP) and vasoactive Peptide (VIP) serum levels are improved, and growth hormone release inhibiting hormone (SS) level declines.RT-PCR experimental results further show, Lactobacillus plantarum YS3 can raise c-Kit, SCF, GDNF mRNA in mice with constipation small intestine and express, and lower TRPV1, NOS mRNA expression. The Lactobacillus plantarum YS3 of high concentration shows more preferable effect, and is significantly better than the lactobacillus bulgaricus of same concentration.These realities Testing result proves, Lactobacillus plantarum YS3 can effectively suppress constipation, can be used in preparing the food of Constipation.
The beneficial effects of the present invention are:The invention provides a kind of Lactobacillus plantarum YS3, it can effectively suppress just It is secret, and effect is significantly better than conventional lactobacillus bulgaricus, can be used in preparing the food of Constipation, not only expands plant Thing lactobacillus YS3 application, improves its application value, and bring new hope to the prevention of constipation.
Brief description of the drawings
Fig. 1 is the agarose gel electrophoresis figure of Lactobacillus plantarum YS3 genomic DNA, and wherein M is λ DNA/Hind III Marker, 1 is Lactobacillus plantarum YS3 genomic DNA.
Fig. 2 is the PCR amplifications of Lactobacillus plantarum YS3 16S rDNA genetic fragments, and wherein M is DNA Marker, C It is the pcr amplification product of Lactobacillus plantarum YS3 16S rDNA genetic fragments for negative control, 1.
Fig. 3 is influences of the Lactobacillus plantarum YS3 to activated carbon inducing constipation mouse weight.
Fig. 4 is influences of the Lactobacillus plantarum YS3 to activated carbon inducing constipation small mouse's head grain melena efflux time.
Fig. 5 is that Lactobacillus plantarum YS3 is expressed c-Kit and SCF mRNA in activated carbon inducing constipation mouse small intestine tissue Influence.
Fig. 6 is Lactobacillus plantarum YS3 to TRPV1, GDNF and NOS mRNA in activated carbon inducing constipation mouse small intestine tissue The influence of expression.
Fig. 3 is into Fig. 6, and LB represents lactobacillus bulgaricus, and LP-YS3 represents Lactobacillus plantarum YS3.
Embodiment
In order that the object, technical solutions and advantages of the present invention are clearer, below in conjunction with accompanying drawing to the excellent of the present invention Embodiment is selected to be described in detail.
The separation and identification of embodiment 1, Lactobacillus plantarum YS3
First, lactic acid bacteria isolates and purifies
Gather 10 parts of the spontaneous fermentation yak yogurt sample of Yushu district, Qinghai Tibetan autonomous prefecture herdsman family, every part of sample nothing After bacterium glass rod is fully stirred evenly, draw 50mL and be put into sterile centrifugation tube, be placed in food sampling box and take back laboratory, 4 DEG C of refrigerators Preserve.
Every part of yak yogurt sample takes 1mL, and 10 times of gradient dilutions are made to 10 with sterile saline-7.Take 10-5、10-6、 10-7Each 1mL dilutions of three dilution factors respectively at equipped with 15mL MRS culture mediums (containing mass ratio be 5% CaCO3) plate In, mix, put 30 DEG C of 72 ± 3h of insulating box culture.Each three, dilution factor work is parallel.After bacterium colony is formed, its form is observed special Levy, the different bacterium colonies that picking has molten calcium circle are inoculated in degreasing milk medium respectively, put 30 DEG C of 24~48h of culture, treat that bacterial strain is given birth to After length is good, streak inoculation puts 30 DEG C of culture 48h in MRS agar mediums;Above step is repeated at least three times, until obtaining Pure bacterium colony.
Pure bacterial strain is rule on MRS solid mediums, 30 DEG C of culture 48h are put, with 10 times of amplification sem observation bacterium colony sizes, The appearance features such as shape, carry out Gram's staining, and the typical bacterial strain of picking form carries out catalase test.Gram is contaminated Color is positive, catalase test is negative meningitidis strains and bacillus strain, are fixed tentatively as lactic acid bacteria.
2nd, the identification of Bacillus acidi lactici
By the lactic acid bacillus mycopremna that preliminary screening goes out carry out respectively growth temperature experiment (10 DEG C, 15 DEG C, 45 DEG C, 60 DEG C, 30min), pH value gradient test, mobility test, catalase test, oxidase test, hydrogen sulfide production test, gelatin liquefaction experiment, Nitrate reduction test, indole test, the experiment of glucose aerogenesis, benzidine test, litmus milk tests, arginine production ammonia examination Test, casein decomposition run and various sugar alcohol fermentation tests.
As a result isolated from 10 parts of yak yogurt samples and be purified into 20 plants of Bacillus acidi lacticis, be Gram-positive, Catalase test is negative.By morphological observation and physiological and biochemical test, this 20 plants of bacterial strains are initially identified as 7 kinds of lactic acid Bacillus, number consecutively is Lb1~Lb7, wherein, numbering is Lb6 Bacillus acidi lactici is initially identified as Lactobacillus plantarum (Lactobacillus plantarum) (table 1).
The numbering of table 1 is LbThe morphological feature and physiological and biochemical test result of 6 Bacillus acidi lactici
Note:+, it is positive;-, it is negative;+ w, weakly positive.
3rd, the identification of Lactobacillus plantarum
It is L to take numberingbSupernatant is abandoned in the bacteria suspension of 6 Bacillus acidi lactici Liquid Culture, centrifugation, thalline is collected, according to bacterium base Because a group DNA extraction kit (Beijing Suo Laibao scientific & technical corporation) methods described extracts genomic DNA, enter row agarose gel electrophoresis (gel mass concentration ratio 0.8%, voltage 80V, electrophoresis 80min), after GelRed nucleic acid staining dyes, in gel imaging system Middle observation (Fig. 1).The genomic DNA of extraction is diluted 200 times, concentration purity testing is carried out with ultraviolet specrophotometer.Using Primer 2 7F:5 '-AGAGTTTGATCCTGGCTCAG-3 ' (SEQ ID No.1) and 1492R:5’- GGCTACCTTGTTACGACTT-3 ' (SEQ ID No.2) enters performing PCR amplification (Fig. 2) to 16S rDNA genetic fragments.Amplification production Thing is sequenced by Sangon Biotech (Shanghai) Co., Ltd. and Beijing Liuhe Huada Genomics Technology Co., Ltd, and will Sequencing result (SEQ ID No.3) is compared with correlated series in Genbank.As a result show, numbering is Lb6 Bacillus acidi lactici The similitude of 16S rDNA gene fragment orders and Lactobacillus plantarum strain CIP 103151 reach 99%.
Therefore, numbering is Lb6 Bacillus acidi lactici is accredited as Lactobacillus plantarum (Lactobacillus plantarum), will It is named as YS3, and is preserved in China typical culture collection center (abbreviation CCTCC, address on December 14th, 2016:Lake No. 299 Wuhan Universitys of Bayi Road of Bei Sheng wuchang, wuhan area), deposit number is CCTCC NO:M2016749.
The prevention effect of embodiment 2, Lactobacillus plantarum YS3 to activated carbon inducing mouse constipation
By making gastrointestinal tract mucous surface be adhered to by activated carbon to mouse stomach activated carbon, cause in alimentary canal under water content Drop, digestive juice is reduced, and triggers gastrointestinal motility to slow down to form constipation.In the early-stage Study to lactic acid bacteria constipation preventive effect, hair A person of good sense confirms that first grain melena defecation time, MTL, Gas, ET, SS, AChE, SP, VIP level etc. all can be as evaluation lactic acid bacterias The index of constipation prevention effect;Detect that correlation mRNA expression can be more true in colon by further molecular biology experiment The physiological function of the confirmation lactic acid bacteria cut.
Main material, instrument and the experimental animal that the present embodiment is used are as follows:Lactobacillus plantarum YS3 (preserving numbers:CCTCC NO:M2016749), lactobacillus bulgaricus (Chinese industrial Microbiological Culture Collection administrative center);MTL、Gas、ET、SS、 AChE, SP and VIP (Beijing pul great achievement bio tech ltd);Trizol reagents, OligodT18、RNase、dNTP、MLV Reverse transcriptase (Invitrogen companies of the U.S.);ROX reference Dye and SYBR Premix Ex Taq II (Japan TAKARA companies);RT-PCR primer (Beijing Tiangeng biochemical technology Co., Ltd).D550 digital cameras (Japan Canon); SevenEasy pH meters (Mettler Toledo Inc. of Switzerland);(U.S. Bole is public for iMark ELIASAs, T100 grads PCRs instrument Department).Seven week old female KM mouse are purchased from Medical University Of Chongqing's Experimental Animal Center (credit number:SYXK (Chongqing) 2012- 0001), raise in temperature and humidity control environment (25 ± 2 DEG C of temperature, relative humidity 50 ± 5%), light dark cycles are adjusted per 12h, are carried For mouse standard feed and drinking-water.
First, the anti-hydrochloric acid in gastric juice abilities of Lactobacillus plantarum YS3 and the detection of cholate tolerance
Lactic acid bacteria will play probiotic action, it is necessary under the strong acidic condition of stomach and enteron aisle, arrive at (typically Large intestine) it is colonized and plays its physiological function.Therefore, in order to study the potential probiotic action of lactic acid bacteria, external virtual mould is set up Type, the anti-hydrochloric acid in gastric juice ability and cholate tolerance for detecting lactic acid bacteria are important detection means.
It is 3.0 to be dissolved in distilled water and adjust pH with 1mol/L HCl NaCl and pepsin, reaches NaCl quality It is 0.35% than the mass ratio for 0.2%, pepsin, vacuum filter removes bacterium, standby as simulated gastric fluid.By plant breast Cultivated after bacillus YS3 activation, take nutrient solution 5mL, 3000r/min centrifugation 10min, thalline is collected, with 5mL physiological saline weights After outstanding, mixed at 1: 9 by volume with simulated gastric fluid, 37 DEG C of culture 3h determine Lactobacillus plantarum YS3 viable counts during 0h and 3h, Survival rate (%)=3h viable count (CFU/mL)/0hs of the Lactobacillus plantarum YS3 in the simulated gastric fluids of pH 3.0 is calculated by formula to live Bacterium number (CFU/mL) × 100.
It is 0.0,0.3%, 0.5%, 1.0% that Lactobacillus plantarum YS3 is inoculated in containing mass ratio by 2% inoculum concentration The MRS-THIO culture mediums of bovine bile, 37 DEG C of culture 24h, using non-inoculated plant lactobacillus YS3 blank cultures as control, Absorbance is determined at 600nm, survival rates (%) of the Lactobacillus plantarum YS3 in cholate=contain bile salt culture-medium is calculated by formula OD600/ blank cultures OD600×100。
As a result as shown in table 2, survival rates of the Lactobacillus plantarum YS3 in the simulated gastric fluids of pH 3.0 and in various concentrations cholate Lactobacillus bulgaricus is above, and survival rates of the Lactobacillus plantarum YS3 in the simulated gastric fluids of pH 3.0 is bulgarian milk bar About 3 times of bacterium, and the survival rate in various concentrations cholate is about 8-11 times of lactobacillus bulgaricus, illustrates Lactobacillus plantarum YS3 has the anti-hydrochloric acid in gastric juice ability and cholate tolerance more stronger than lactobacillus bulgaricus.
The anti-hydrochloric acid in gastric juice abilities of the Lactobacillus plantarum YS3 of table 2 and the detection of cholate tolerance
2nd, prevention effects of the Lactobacillus plantarum YS3 to activated carbon inducing mouse constipation
Choose body weight about 25g kunming mice 100, be equally divided into 5 groups, be normal group, constipation control group respectively, protect plus Leah lactobacillus treatment group, Lactobacillus plantarum YS3 low concentrations treatment group and high concentration treatment group.Adapt to after raising 1 week, normal group Diet is normally freely taken in constipation control group mice and is drunk water 2 weeks;Except normally freely taking in diet and drinking-water in this is 2 weeks Outside, the 1.0 × 10 of the daily every gavage 2mL of lactobacillus bulgaricus treatment group mouse9CFU/mL lactobacillus bulgaricus, plants The 1 × 10 of thing lactobacillus YS3 low concentrations treatment group and the daily every difference gavage 2mL of high concentration treatment group mouse8CFU/mL's Lactobacillus plantarum YS3 and 1 × 109CFU/mL Lactobacillus plantarum YS3.After 2 weeks in addition to normal group mouse, other 4 groups of mouse are every (temperature is 4 DEG C to the activated carbon water of day gavage 2mL, and 10% activated carbon is added into the water containing 10% gum arabic in mass ratio Suspension is made in solution), while lactic acid bacteria processing each group continues gavage lactic acid bacteria, continue 3 days.
1st, influences of the Lactobacillus plantarum YS3 to mouse weight and excrement
In experimentation the timing of daily morning determine the body weight of all mouse, dietary intake, amount of drinking water, stool weight and Excrement humidity.
Changes of weight is the important indicator that constipation occurs for mouse.As seen from Figure 3, in first 2 weeks each group mouse body weight All showing between normal growth state, each group does not have notable difference;After activated carbon inducing constipation, not by activated carbon at The normal group mouse weight of reason continues to increase, and remaining each group mouse weight declines, wherein the body of constipation control group mice It is reduced by up to again, the body weight of Lactobacillus plantarum YS3 high concentration treatment group mouse is significantly higher than the guarantor of same concentration closest to normal group Plus Leah lactobacillus treatment group, illustrate that Lactobacillus plantarum YS3 can suppress the mouse weight decline that constipation is caused, and effect is better than Lactobacillus bulgaricus.
Defecation state most can significantly embody stool weight, granule number and water content in the degree of constipation, constipation state It is important excrement state index, these index decreaseds show the exacerbation of constipation degree.As shown in table 3, the 1-14 days it is each Stool weight, granule number and the water content of group mouse are not significantly different;After activated carbon inducing constipation, the 15-17 days normal groups Stool weight, granule number and the water content highest of mouse, constipation control group are minimum, lactobacillus bulgaricus and Lactobacillus plantarum YS3 can stool weight that significantly relief of constipation is caused, granule number and water content decline (P < 0.05), but Lactobacillus plantarum Stool weight, granule number and the water content of YS3 high concentration treatment group mouse are closest to normal group, and the guarantor for being significantly higher than same concentration adds Leah lactobacillus treatment group, illustrates that Lactobacillus plantarum YS3 can suppress the stool in mice weight that constipation is caused, granule number and aqueous Amount declines, and effect is better than lactobacillus bulgaricus.
Influences of the Lactobacillus plantarum YS3 of table 3 to stool in mice
Note:Alphabetical different expression group differences are significantly (P < 0.05).
2nd, influences of the Lactobacillus plantarum YS3 to activated carbon propulsion rate and first grain melena efflux time
To the jejunitas 24h of all mouse implementations including normal group after 17th day gavage activated carbon water, but still give mouse Freely take in the activated carbon water of all every gavage 0.2mL of mouse after drinking-water, 24h;Every group of half mouse (10) is real after 30min Apply disconnected neck to put to death, take mice plasma standby, while taking small intestine to observe propulsion rate of the activated carbon in mouse small intestine, propulsion rate (%) =activated carbon promotes length/total small intestinal length × 100;Every group of remaining 10 mouse continue to observe its discharge first grain melena when Between.
Constipation causes the reduction of intestines peristalsis frequency, and excrement rests on the time increase of enteron aisle, causes harmful bacteria with excrement Continuously bred for food, so as to threaten intestinal health, aggravate constipation.Activated carbon is promoted in small intestine after activated carbon inducing mouse constipation Length and activated carbon propulsion rate can be used as the index for evaluating small intestine activity and constipation degree.As can be seen from Table 4, activated carbon is lured Lead after constipation, lactobacillus bulgaricus and Lactobacillus plantarum YS3 can promote propulsion length of the activated carbon in mouse small intestine, But the activated carbon propulsion rate highest of Lactobacillus plantarum YS3 high concentration treatment groups, is significantly higher than the processing of Lactobacillus plantarum YS3 low concentrations Group and the lactobacillus bulgaricus treatment group (P < 0.05) with concentration, illustrate that Lactobacillus plantarum YS3 can improve activated carbon small Propulsion rate in mouse small intestine, and effect is better than lactobacillus bulgaricus.
The influence that the Lactobacillus plantarum YS3 of table 4 is promoted to mice with constipation intestines and stomach
Note:Alphabetical different expression group differences are significantly (P < 0.05).
Constipation causes intestinal movement to slow down, and excrement holdup time in enteron aisle increases, and lengthens first grain melena efflux time, Shorter first grain melena efflux time means that intestinal movement is more normal.As seen from Figure 4, the first grain melena discharge of normal group Time is most short (83 ± 9min), and the first grain melena efflux time of constipation control group is most long (195 ± 28min), Lactobacillus plantarum YS3 The first grain melena efflux time (112 ± 19min) of high concentration treatment group is considerably shorter than Lactobacillus plantarum YS3 low concentration treatment groups (147 ± 26min) and the lactobacillus bulgaricus treatment group (163 ± 22min) (P < 0.05) with concentration, illustrates the newborn bar of plant Bacterium YS3 can reduce the first grain melena efflux time of mice with constipation, with good constipation mitigation, and effect better than protecting plus Leah lactobacillus.
3rd, influences of the Lactobacillus plantarum YS3 to mice serum MTL, Gas, ET, SS, AChE, SP and VIP level
The mice plasma of collection is taken, 4500rmp centrifugation 15min separate serum, mouse blood is determined by kit operating instruction Clear MTL, Gas, ET, SS, AChE, SP and VIP level.MTL can stimulate the generation of pepsin and promote intestinal movement; Gas can promote gastrointestinal secretion and gastrointestinal motility, promote pylorus relaxation, play a part of relief of constipation;ET is stable in antiotasis Property and maintain to play an important role in basic cardiovascular system;SS can stimulate intestinal movement;AChE can adjust contraction of muscle and Mucous secretion, can make loosening all muscles to promote excrement to discharge;SP is a kind of to contribute to the material of intestines peristalsis;Keep VIP Normal contents in intestinal wall are also to stablize the important means of function of intestinal canal.
As can be seen from Table 5, the horizontal highest of MTL, Gas, ET, AChE, SP and VIP of normal group mice serum, SS levels It is minimum;Constipation control group mice shows opposite trend;Lactobacillus bulgaricus and Lactobacillus plantarum YS3 can make above-mentioned The serum levels of material are close to normal group level, but Lactobacillus plantarum YS3 effect is more notable, illustrate that Lactobacillus plantarum YS3 can So that MT in serum L, Gas, ET, SS, AChE, SP and VIP level of mice with constipation is tried one's best, holding is normal, so that relief of constipation.
Influences of the Lactobacillus plantarum YS3 of table 5 to mice serum MTL, Gas, ET, SS, AChE, SP and VIP level
Note:Alphabetical different expression group differences are significantly (P < 0.05).
4th, the influence that Lactobacillus plantarum YS3 is expressed mouse small intestine tissue mRNA
The small intestine of mouse is taken, colon's total serum IgE is extracted with RNAzol after crushing, is diluted to 1 μ g/ μ L;Take 2 μ L dilute Total RNAs extraction liquid after releasing, sequentially adds 1 μ L oligodT18, RNase, dNTP, MLV enzyme and 10 μ L 5 × buffer, 37 DEG C 120min, 99 DEG C of 4min, 4 DEG C of 3min synthesis cDNA;Then use primer described in table 6, RT-PCR amplification c-Kit, SCF, TRPV1, GDNF and NOS mRNA expression, internal reference is used as using housekeeping gene GAPDH;Finally with containing the bromination that mass ratio is 1% The agar electrophoresis of second ingot checks pcr amplification product.
The RT-PCR primer sequence of table 6
ICC (Cajal interstitial cells) in constipation patient enteron aisle is less, and c-Kit is ICC Specific marker, is ICC The key of propagation.SCF concentration is extremely important to ICC propagation, and ICC can not be grown in the environment of SCF is not present.Document report ICC contents in road, mice with constipation colon are less, and the c-Kit expressions in colon also decline.Lactobacillus plantarum YS3 mouse small intestine tissue c-Kit and SCF the mRNA influences expressed are shown in Fig. 5, constipation control group mice small intestine c-Kit and SCF mRNA expression declines, and Lactobacillus plantarum YS3 can significantly raise c-Kit the and SCF mRNA expression in mouse small intestine, and Acted on during high concentration more obvious.
TRPV1 with defecation it is verified that have substantial connection, and the release of neurotransmitter can be triggered by activating TRPV1, so that Cause intestinal motility obstacle.TRPV1 expression increases are a notable phenomenons of damage of intestines, because gastrointestinal disorder causes enteron aisle to damage Wound, makes constipation patient have higher TRPV1 expression.GDNF can adjust the function of gangliocyte, thus contribute to repair by The enteron aisle of damage, can prevent constipation.NOS plays an important role to Gastrointestinal motility adjustment.NO increase can cause more serious colon to move Power obstacle, control NOS can reduce NO content, be the feasible way for controlling constipation.Lactobacillus plantarum YS3 is to mouse small intestine group Fig. 6 is shown in the influence for knitting middle TRPV1, GDNF and NOS mRNA expression, and normal group mouse has highest GDNF mRNA expression and most Low TRPV1, NOS mRNA expression, and the GDNF mRNA of constipation control group mice express minimum, TRPV1 and NOS mRNA tables Up to highest, relative to constipation control group, lactobacillus bulgaricus and Lactobacillus plantarum YS3 can raise GDNF mRNA expression Expressed with TRPV1, NOS mRNA is lowered, but Lactobacillus plantarum YS3 effect is substantially better than lactobacillus bulgaricus.
Finally illustrate, the above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, although pass through ginseng According to the preferred embodiments of the present invention, invention has been described, it should be appreciated by those of ordinary skill in the art that can So that various changes are made to it in the form and details, the present invention limited without departing from appended claims Spirit and scope.
<110>The college of education of Chongqing second
<120>Lactobacillus plantarum YS3 and its application in the food of Constipation is prepared
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<212>DNA
<213>Lactobacillus plantarum YS3(Lactobacillus plantarum YS3)
<220>
<223>16S rDNA genetic fragments
<400>3
taatacatgc aagtcgaacg aactctggat tgattggtgc ttgcatcatg atttacattt 60
gagggagtgg cgaacgtgca atggcggctt tggtgagtaa cacgtgggaa acctgcccag 120
aagcggggaa taacacctgg aaacagatgc tataccgcat aacaacttgg accgcatggt 180
ccgagnttga aagatggctt cggctatcac ttttggatgg tcccgcggcg tattagctag 240
atggtggggt aacggctcac catggcaatg atacgtagcc gacctgagag ggtaatcggc 300
cacattggga ctgactcacg gccaaactcc tacgggaggc agcagtaggg aatcttccac 360
aatggacgaa gtctgatgga gcaacgccgc gtgagtgaaa agggtttcgg ctcgtaaact 420
ctgttgttaa agaagaacat atctgagagt aactgttcag gtattgacgg tatttaacag 480
aaagccacgg ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttgtcc 540
ggatttattg ggcgtaaagc gagcgcaggc ggttttttaa gtctgatgtg aaagccttcg 600
gctcaaccga agaagtgcat cggaaatggg aaacttgagt gcagaagagg acagtggaac 660
tccattgtag cggtgaaatg cgtagatata tggaagaaca ccagtggcga aggcgctgtc 720
tggtctgtaa ctgacgctga ggctcgaaag tatggtagca aacaggatta gataccctgg 780
tatccatacc gtaaacgatg aatgctaagt gttggagggt ttccgccctt cagtgctgca 840
gctaacgcat taagcattcc gcctggggag tacggccgca aggctgaaac tcaaaggaat 900
tacgggggcc cgcacaagcg gtggagcatg tggttaaatt cgaagctacg cgaagaacct 960
taccaggtct tgacatacta tgcaaatcta agagattaga cgttcccttc ggggacatgg 1020
atacaggtgg tgcatggttg tcgtcagctc atgtcgtgag atgttgggtt agtcccgcac 1080
gagcgcaacc cttattatca gttgccagcg ttaagttggg cactctggtg agactgccgg 1140
tgacaaacgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acctgggcta 1200
cacacgtgct acaatggatg gtacaacagt tgcgaactcg cgagagtaag ctaatctctt 1260
aaagccattc tcagttcgga ttgtaggctg caactcgcct acatgaagtc ggaatcgcta 1320
gtaatccgga tcagcatgcc gcggtgaata cgttcccggg ccttgacaca ccgcccgtca 1380
caccatgaga gtttgtaaca cccaaagtcg gtggggtaac cttttaggaa ccagccgcct 1440
aagggggaca gtagagtaag gtggat 1466
<210>4
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification c-Kit mRNA expression
<400>4
agaccgaacg caactt 16
<210>5
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification c-Kit mRNA expression
<400>5
ggtgccatcc acttca 16
<210>6
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification SCF mRNA expression
<400>6
aaactggtgg cgaatc 16
<210>7
<211>16
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification SCF mRNA expression
<400>7
cacgggtagc aagaac 16
<210>8
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification TRPV1 mRNA expression
<400>8
agcgagttca aagacccaga 20
<210>9
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification TRPV1 mRNA expression
<400>9
ttctccacca agagggtcac 20
<210>10
<211>19
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification GDNF mRNA expression
<400>10
ttttattcaa gccaccatc 19
<210>11
<211>17
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification GDNF mRNA expression
<400>11
agcccaaacc caagtca 17
<210>12
<211>21
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification NOS mRNA expression
<400>12
ccacatctgg caggatgaga a 21
<210>13
<211>21
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification NOS mRNA expression
<400>13
aggcacagaa ctgagggtac a 21
<210>14
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The sense primer of RT-PCR amplification GAPDH mRNA expression
<400>14
cggagtcaac ggatttggtc 20
<210>15
<211>20
<212>DNA
<213>Artificial sequence
<220>
<223>The anti-sense primer of RT-PCR amplification GAPDH mRNA expression
<400>15
agccttctcc atggtcgtga 20

Claims (2)

1. Lactobacillus plantarum YS3 (Lactobacillus plantarum YS3), deposit number is CCTCC NO:M2016749.
2. applications of the Lactobacillus plantarum YS3 in the food of Constipation is prepared described in claim 1.
CN201710542452.6A 2017-07-05 2017-07-05 Lactobacillus plantarum YS3 and its application in the food of Constipation is prepared Pending CN107099491A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109700033A (en) * 2019-03-04 2019-05-03 山东环亿生物科技有限公司 A kind of adjusting enteron aisle, probiotic composition of prevent constipation and preparation method thereof
CN110692726A (en) * 2018-07-10 2020-01-17 重庆第二师范学院 Lactobacillus plantarum CQPC01 and application thereof in preparation of food for improving constipation
CN110699271A (en) * 2018-07-10 2020-01-17 重庆第二师范学院 Lactobacillus plantarum CQPC02 and application thereof in preparation of food for improving constipation

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106906165A (en) * 2017-04-25 2017-06-30 重庆第二师范学院 A kind of Lactobacillus plantarum and its application in the food for preparing Constipation

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106906165A (en) * 2017-04-25 2017-06-30 重庆第二师范学院 A kind of Lactobacillus plantarum and its application in the food for preparing Constipation

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110692726A (en) * 2018-07-10 2020-01-17 重庆第二师范学院 Lactobacillus plantarum CQPC01 and application thereof in preparation of food for improving constipation
CN110699271A (en) * 2018-07-10 2020-01-17 重庆第二师范学院 Lactobacillus plantarum CQPC02 and application thereof in preparation of food for improving constipation
CN110699271B (en) * 2018-07-10 2022-11-01 善恩康生物科技(苏州)有限公司 Lactobacillus plantarum CQPC02 and application thereof in preparation of food for improving constipation
CN109700033A (en) * 2019-03-04 2019-05-03 山东环亿生物科技有限公司 A kind of adjusting enteron aisle, probiotic composition of prevent constipation and preparation method thereof
CN109700033B (en) * 2019-03-04 2021-06-04 山东环亿生物科技有限公司 Probiotic composition for regulating intestinal tract and preventing constipation and preparation method thereof

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Application publication date: 20170829