CN111436203B - Fermented lactobacillus plantarum and application thereof - Google Patents

Fermented lactobacillus plantarum and application thereof Download PDF

Info

Publication number
CN111436203B
CN111436203B CN201880030861.0A CN201880030861A CN111436203B CN 111436203 B CN111436203 B CN 111436203B CN 201880030861 A CN201880030861 A CN 201880030861A CN 111436203 B CN111436203 B CN 111436203B
Authority
CN
China
Prior art keywords
lactobacillus plantarum
fermented
cgmcc
fermentation
products
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201880030861.0A
Other languages
Chinese (zh)
Other versions
CN111436203A (en
Inventor
李慧
陈历水
高嘉星
刘蕾
刘洋
应欣
史玉莹
张连慧
倪伟
钱承敬
王勇
王宇
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cofco Nutrition and Health Research Institute Co Ltd
Original Assignee
Cofco Nutrition and Health Research Institute Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cofco Nutrition and Health Research Institute Co Ltd filed Critical Cofco Nutrition and Health Research Institute Co Ltd
Publication of CN111436203A publication Critical patent/CN111436203A/en
Application granted granted Critical
Publication of CN111436203B publication Critical patent/CN111436203B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • 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
    • 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
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus
    • C12R2001/25—Lactobacillus plantarum

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Dairy Products (AREA)

Abstract

The invention belongs to the technical field of microbial fermentation, and particularly relates to Lactobacillus plantarum and application thereof, wherein the Lactobacillus plantarum is classified and named Lactobacillus plantarum and is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, and the preservation number of the Lactobacillus plantarum is CGMCC No. 16441. The lactobacillus plantarum disclosed by the invention has the capabilities of acid resistance and cholate resistance, has very high adhesion to intestinal epithelial cell strain HT-29 cells, has very high acid production and polysaccharide production capabilities, and has a higher acid production speed, a stronger acid production capability and a higher polysaccharide production capability compared with other lactic acid bacteria. Fermented dairy products, fermented grain beverages, fermented bean products, fermented fruit and vegetable juices and fermented feeds prepared by the lactobacillus plantarum have unique flavor, uniform texture, smooth mouthfeel and good sensory characteristics. Can be widely applied to the field of fermentation products and can also be used for preparing live bacteria preparations.

Description

Fermented lactobacillus plantarum and application thereof
Technical Field
The invention relates to the technical field of microbial fermentation, and particularly discloses lactobacillus plantarum, a direct vat set starter containing the lactobacillus plantarum and a preparation method thereof, and application of the lactobacillus plantarum in the fields of preparing fermented dairy products, fermented grain beverages, fermented bean products, fermented fruit and vegetable products and fermented feeds, and also discloses application of the lactobacillus plantarum in preparing live bacterial preparations and the prepared live bacterial preparations.
Background
Lactic Acid Bacteria (LAB) are a general term for a group of bacteria that can utilize fermentable carbohydrates to produce large amounts of lactic acid. This is a relatively bulky group of bacteria, which can be currently divided into at least 18 genera, with a total of over 200 species, and a large variety of species. The traditional fermented dairy products in Qinghai, Tibet, inner Mongolia, Xinjiang and other places of China contain rich lactobacillus strain resources, and are rich probiotics strain treasury.
Lactobacillus plantarum strains among lactic acid bacteria are widely used in food production, especially in fermented foods. There are studies showing that: the acid production capability and the fermentation flavor characteristics of the lactic acid bacteria play an important role in the production of fermented milk products, fermented cereal beverages, fermented bean products, fermented fruit products and fermented feeds, and can be used alone or in combination with other bacteria. Lactic acid bacteria as potential probiotics need to have the following properties: has outstanding probiotic function, is required to be resistant to gastrointestinal tract acid environment and bile salt environment, and has better cell adhesion.
The main roles of lactic acid bacteria in fermented products are acid production and flavor generation. The acid-producing characteristics of lactic acid bacteria are important characteristics for screening a fermentation product starter. Lactic acid bacteria having a strong acid-producing ability generally have a strong tendency to acidify thereafter. Lactic acid bacteria with fast acid production and large acid production amount can bring lower fermentation acidity of final products, which is an effective measure for ensuring low post-acidification (such as dairy products). As probiotics, gastric juice and intestinal juice form a biological barrier of a human body, and lactobacillus can only pass through the gastrointestinal tract and attach DIAN to epithelial cells of the intestinal tract to realize colonization so as to exert beneficial effects on the human body or animal hosts.
Exopolysaccharides (EPS) of lactic acid bacteria are polysaccharides produced by lactic acid bacteria during growth metabolism and secreted outside the cells, released into the growth substrate or capsular polysaccharides tightly bound to the cell surface. The functions of the following aspects are mainly realized: the quality of the product is improved, the product is fine and uniform, and the smooth mouthfeel is increased; the water retention is increased, the viscosity of the product is improved, the non-specific adhesion of the lactobacillus to the intestinal surface is improved, the chance of field planting of the lactobacillus is increased, and a certain promotion effect on the regulation of the intestinal microbial flora is achieved.
Therefore, the compound microbial compound has good acid production capacity and exopolysaccharide production capacity, can resist gastrointestinal tract stress, has good cell adhesion, and has prominent fermentation sensory characteristics which are necessary factors for screening leavening agents and probiotics.
Disclosure of Invention
In order to solve the technical problems, the invention screens the lactobacillus plantarum which has high acid production speed, stable acid production and stable curd genetic property and unique fermented product flavor, and the lactobacillus plantarum is used for fermented dairy products, grain products, bean products, fruit and vegetable products and feed products to improve the quality of the fermented products. The lactobacillus plantarum can also tolerate the acid environment of the gastrointestinal tract, can produce extracellular polysaccharide with high yield, improves the field planting property of cells, is used for preparing a probiotic viable bacteria preparation, and is beneficial to adjusting the composition of intestinal microbial flora.
In order to achieve the above objects, in a first aspect, the present invention provides a Lactobacillus plantarum, which is classified and named Lactobacillus plantarum, and is deposited in the china general microbiological culture collection center of the committee for culture collection of microorganisms, with the collection number of CGMCC No. 16441.
In a second aspect, the present invention provides the use of lactobacillus plantarum as described above in the field of preparation of fermented dairy products, fermented cereal beverages, fermented soy products, fermented fruit delicatessens, fermented feed.
In a third aspect, the present invention provides the use of a lactobacillus plantarum as described above for the preparation of a ready-to-use starter culture.
In a fourth aspect, the present invention provides a direct vat set starter, which is prepared by fermenting and culturing the lactobacillus plantarum mentioned above in a fermentation medium.
In a fifth aspect, the present invention provides a process for the direct vat set fermentation as described above, the process comprising:
(1) will be as described aboveThe lactobacillus plantarum is subjected to fermentation culture in a fermentation culture medium, and the viable count is 108More than cfu/mL;
(2) centrifuging the zymocyte liquid obtained in the step (1), washing with buffer solution, adding freeze-drying protective agent, and adjusting viable bacteria concentration to 10 10More than cfu/mL, and after being uniformly mixed, the mixture is subjected to vacuum freeze drying to obtain the direct vat set starter.
In a sixth aspect, the present invention provides the use of the direct vat set as described above and/or the direct vat set prepared by the method as described above in the field of preparing fermented dairy products, fermented cereal beverages, fermented soy products, fermented fruit boiled products, fermented feed.
In a seventh aspect, the present invention provides the use of a lactobacillus plantarum as described above, for the preparation of a live bacterial formulation.
In an eighth aspect, the invention provides a viable bacteria preparation, which is prepared by fermenting and culturing the lactobacillus plantarum in a fermentation medium.
The invention has the beneficial effects that:
1. safe and healthy, and low cost: the lactobacillus plantarum CGMCC No.16441 is a safe strain which is screened from the milk cakes made by the traditional method in the herdsman in inner Mongolia and can be used for food, and the method has no chemical addition, is green and natural, and is nutritional and healthy;
2. the lactobacillus plantarum CGMCC No.16441 can effectively improve the fermentation quality of fermented dairy products, cereal products, bean products, fruit and vegetable products and feed products and exert the probiotic function of probiotics; the lactobacillus plantarum CGMCC No.16441 is used as a leavening agent, so that the acidity of the product can be effectively improved, the content of polysaccharide is increased, the nutritional value and flavor characteristics of the product are improved, and the sensory quality characteristics of the product are improved to a certain extent; the lactobacillus plantarum CGMCC No.16441 is used as probiotics, so that the colonization of cells can be effectively improved;
3. The lactobacillus plantarum CGMCC No.16441 is used as a leavening agent and a probiotic viable bacteria preparation, is easy to culture and prepare, and has high viable bacteria concentration.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Biological preservation
The Lactobacillus plantarum provided by the invention is preserved in the general microbiological center of China Committee for culture Collection of microorganisms (CGMCC) in 2018, 9 and 7, and the preservation number is CGMCC No.16441, the preservation address is No. 3 of Beijing university Hokko No.1 of sunward areas, and the institute of microbiology (CGMCC for short).
Drawings
In order that the present disclosure may be more readily and clearly understood, the following detailed description of the present disclosure is provided in connection with specific embodiments thereof and the accompanying drawings, in which,
FIG. 1 shows the colony morphology of Lactobacillus plantarum of the invention on a culture medium;
FIG. 2 shows the microscopic morphology of Lactobacillus plantarum in accordance with the present invention;
FIG. 3 is a graph showing the pH value dynamic change of acid produced by different strains according to the present invention;
FIG. 4 shows the analysis and detection results of the fermented flavor of Lactobacillus plantarum in accordance with the present invention.
Detailed Description
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In a first aspect, the invention provides a Lactobacillus plantarum, which is classified and named Lactobacillus plantarum, is deposited in the China general microbiological culture Collection center (CGMCC), and has a preservation number of CGMCC No. 16441.
The lactobacillus plantarum CGMCC No.16441 has the following properties:
(1) morphological characteristics: on an MRS culture medium, colonies are milky white, round, smooth in surface, moist and opaque, and cells under a microscope are rod-shaped and gram-positive;
(2) genetic stability: the acid production and curd performance are stable heredity, the subculture is carried out for 10 generations, the acidity value is 89.74, and the consistency reaches 4721.86/g;
(3) acid generation: acid production capability test is carried out according to the MC liquid culture medium, the obtained strain CGMCC NO.16441 has pH value of 3.76 after fermentation for 24h, and the acid production capability is higher than that of a commercial strain;
(4) fermentation flavor characteristics: besides common flavor substances, the fermented product also contains 3-hydroxy-2-butanone, 2-heptanone, 2-nonanone, ethyl caproate and other components, so that the product is endowed with faint scent flavor;
(5) in vivo tolerance: the survival rate after 3h of treatment in artificial gastric juice with pH 3.0 is 62%; survival rate of treatment in simulated intestinal fluid for 24h was 39%; shows good gastrointestinal tract tolerance;
(6) Polysaccharide production capacity: the strain is inoculated in an MRS liquid culture medium, after the strain is cultured for 48 hours at 37 ℃, the extracellular polysaccharide yield reaches 803mg/L which is far higher than 150-600mg/L of the similar strain subjected to optimized culture conditions;
(7) adhesiveness: up to 1000 bacteria/100 cells (HT-29).
The lactobacillus plantarum CGMCC No.16441 is screened from milk cakes made by traditional methods in herdsmen in inner Mongolia regions.
The Lactobacillus plantarum provided by the present invention can produce a large amount of viable cells of Lactobacillus plantarum by liquid culture, and the culture method is not particularly limited as long as the Lactobacillus plantarum can proliferate, and may be, for example, 106- 8Inoculating the live bacteria of the lactobacillus plantarum into a lactobacillus culture medium in the inoculation amount of CFU/mL, and culturing at 15-40 ℃ for 8-72 hours under an anaerobic or aerobic condition to obtain a culture solution. The culture medium for Lactobacillus may be any suitable culture medium known in the art, such as milk and/or "Lactobacillus-biological basis and applications" (Darby Bin)Lactobacillus (MRS) medium as described in light industry press, 1996 press).
In the present invention, the viable cells of lactobacillus plantarum in the culture solution may be further separated, and the method for separating is not particularly limited as long as the cells can be enriched from the culture solution, and for example, the separation can be achieved by centrifugation and/or filtration, and the conditions for centrifugation and filtration may be known conditions, and the present invention is not described herein again.
In a second aspect, the invention provides an application of the lactobacillus plantarum CGMCC No. 16441.
The application of the lactobacillus plantarum CGMCC No.16441 disclosed by the invention refers to the application in fermented food in one embodiment of the invention.
The fermented product, in one embodiment of the invention, comprises dairy products, cereal products, bean products, fruit and vegetable products and feed products.
The dairy product, in one embodiment of the invention, may comprise yogurt, a live lactic acid bacteria beverage, or the like.
The cereal product, in one embodiment of the present invention, may include oat fermented beverage, black rice fermented beverage, buckwheat fermented beverage, and the like.
The soy product, in one embodiment of the present invention, may include fermented soy milk or the like.
The fruit and vegetable product, in one embodiment of the present invention, may comprise fermented fruit and vegetable juice, or the like.
The feed product, in one embodiment of the invention, may comprise a solid anaerobic biological feed.
In the embodiment of the invention, in the conventional production process of producing fermented dairy products, grain products, bean products, fruit and vegetable products and feed products, the lactobacillus plantarum CGMCC No.16441 is inoculated into raw materials to be treated according to the conventional use, and the lactobacillus plantarum CGMCC No.16441 is fermented or survived at the temperature and pressure capable of propagating the lactobacillus plantarum CGMCC No. 16441. By adding CGMCC No.16441 into fermentation substrate, the metabolite of the CGMCC No.16441 ensures that the fermentation product has certain excellent characteristics of acidity, fragrance and the like, prolongs the preservation time of the product, and improves the nutritive value and flavor characteristics of the product.
In a third aspect, the invention provides application of the lactobacillus plantarum CGMCC No.16441 in preparation of a direct vat set starter.
In a fourth aspect, the invention provides a direct vat set starter, which is prepared by fermenting and culturing lactobacillus plantarum CGMCC No.16441 in a fermentation medium.
According to the present invention, the fermentation medium may be any of various media conventionally used in the art for fermentation of lactic acid bacteria, for example, MRS medium as described above, and may be any of various media suitable for fermentation of lactic acid bacteria optimized on the basis of MRS medium.
According to the present invention, the fermentation conditions may be conventional conditions for fermentation culture of lactic acid bacteria well known in the art, and for example, the temperature of the fermentation culture may be 30 to 40 ℃.
According to the present invention, the preparation method of the direct vat set starter can be prepared according to the conventional method in the field, and in one embodiment of the present invention, the direct vat set starter can be prepared according to the following method: culturing Lactobacillus plantarum CGMCC No.16441 until viable count reaches 108More than cfu/mL, and is prepared by vacuum freeze drying after being uniformly mixed.
The direct vat set starter is bacterial powder prepared by vacuum freeze drying lactobacillus plantarum CGMCC No.16441 zymocyte liquid in one embodiment of the invention, and the bacterial powder contains more than 10 hundred million/g of active lactobacillus plantarum CGMCC No. 16441.
In a fifth aspect, the present invention provides a method of preparing a direct vat set as described above, the method comprising:
(1) the lactobacillus plantarum CGMCC No.16441 is subjected to fermentation culture in a fermentation culture medium, and the viable count reaches 108cfu/mL or more;
(2) centrifuging the zymocyte liquid obtained in the step (1), washing with buffer solution, and adding freeze-drying agentProtecting agent, and adjusting viable bacteria concentration to 10 10More than cfu/mL, and after being uniformly mixed, the mixture is subjected to vacuum freeze drying to obtain the direct vat set starter.
The method for fermentation culture of Lactobacillus plantarum CGMCC No.16441 according to the present invention has been described in detail above, and will not be repeated herein in order to avoid unnecessary repetition.
According to the present invention, the method for centrifuging the fermentation broth can be performed according to conventional methods in the art, for example, centrifugation at 5000-12000rpm for 5-20min in a refrigerated centrifuge can be used to obtain cell pellet.
According to the present invention, the lyoprotectant may be various cryoprotectants conventional in the art, and for example, may be at least one of skim milk powder, maltodextrin, trehalose, dextran, and glycerin.
According to the present invention, the buffer may be a buffer conventionally used in the art for washing the bacterial cells, and may be, for example, physiological saline or PBS buffer.
In a sixth aspect, the invention provides the use of the direct vat set as described above and/or the direct vat set prepared by the method as described above in the field of preparing fermented dairy products, fermented cereal beverages, fermented soy products, fermented fruit boiled products, fermented feed.
In a seventh aspect, the invention provides an application of lactobacillus plantarum CGMCC No.16441 in preparing a live bacterium preparation.
According to the present invention, the live bacterial preparation may be in various forms known in the art, such as, but not limited to, capsules, tablets, oral liquids, and powders.
In an eighth aspect, the invention provides a viable bacteria preparation, which is prepared by fermenting and culturing the lactobacillus plantarum in a fermentation medium.
According to the present invention, the preparation method of the live bacterial preparation can be performed by referring to the direct vat set starter as described above to avoid unnecessary repetition, and thus, the details are not repeated herein.
Examples
The media formulations referred to in the following examples are as follows:
MRS medium (Man Rogosa sharp Broth): peptone 1 wt%, beef powder 0.5 wt%, yeast powder 0.4 wt%, glucose 2 wt%, Tween 800.1 wt%, and K2HPO4.7H20.2% by weight of O, sodium acetate 3H20.5% by weight of O, 0.2% by weight of triammonium citrate, MgSO4·7H20.02 wt.% of O, MnSO4·4H20.005 percent of O and 1.5 percent of agar powder by weight are heated and dissolved, the pH value is adjusted to 6.2 +/-0.2, and the mixture is sterilized for 15-20min under high pressure at 121 ℃.
MC culture medium: 0.5 weight percent of soybean peptone, 0.3 weight percent of beef powder, 0.3 weight percent of yeast powder, 2 weight percent of glucose, 2 weight percent of lactose, 1 weight percent of calcium carbonate, 1.5 weight percent of agar and 0.5 weight percent of 1 percent neutral red solution. Adding the above 7 components into distilled water, heating to dissolve, adjusting pH to 6.0 + -0.2, adding neutral red solution, packaging, and autoclaving at 121 deg.C for 15-20 min.
Example 1
This example is used to illustrate the separation, purification and characterization of Lactobacillus plantarum CGMCC No.16441
Strain: the lactobacillus plantarum CGMCC No.16441 is separated from the traditional milk crisp of inner Mongolia and is preserved by the nutritional and health strain preservation center of the Chinese food nutrition and health research institute.
The inventor collects samples such as traditional milk cake, milk powder, milk bar, milk bean curd and the like prepared by the traditional method from herdsmen in the inner Mongolia Hulunbel, Xilin Guo Allen and other areas, and uses sterile physiological saline to carry out gradient dilution to 10 DEG-6MRS plates and MC plates were sequentially applied to each dilution gradient, and cultured at 36. + -. 1 ℃ for 72 hours. And selecting colonies with different colony morphologies by an inoculating needle, and streaking on an MRS plate and an MC plate until single colonies are uniform in size and consistent in morphology.
Strains were selected which were purple in gram stain, catalase negative, and non-sporulating. The strain separated to tentatively become lactobacillus is activated for 3 generations in MRS liquid culture medium and then subjected to physiological and biochemical identification and molecular biological identification, acid production and curding genetic stability, fermentation performance, sensory evaluation, fermentation flavor substances, polysaccharide production capacity, gastrointestinal tract acid environment resistance, gastrointestinal tract bile salt environment resistance and other aspects of lactobacillus are researched, and a lactobacillus plantarum CGMCC NO.16441 is finally screened and obtained from a plurality of wild lactobacillus through a plurality of rounds of research and argumentation.
1. Morphological identification
The screened lactobacillus plantarum CGMCC NO.16441 is cultured for 72 hours at the temperature of 36 +/-1 ℃, and as shown in figure 1, the colony on an MRS culture medium is milky white, round, smooth in surface, moist and opaque. As shown in FIG. 2, the cell morphology of Lactobacillus plantarum CGMCC NO.16441 is rod-shaped and gram-positive under the microscope.
2. Physiological and biochemical identification
The screened strain CGMCC NO.16441 is subjected to physiological and biochemical identification by using a French Merrier API identification system, and the identification result is shown in Table 1 below. Through physiological and biochemical identification, the isolated strain is Lactobacillus plantarum (Lactobacillus plantarum).
TABLE 1 physiological and biochemical identification experiment results of strain CGMCC NO.16441
Figure GWB0000003175010000081
Figure GWB0000003175010000091
[ note ]: in the table, "+" indicates that the biochemical reaction result is positive, and "-" indicates that the biochemical reaction result is negative.
3. Molecular identification
The 16s rDNA of the separated strain is cloned and sequenced, the nucleotide sequence of the 16s rDNA gene is shown as SEQ ID No.1, the 16s rDNA sequence of the strain is compared with the sequence of NCBI Lactobacillus plantarum, and the similarity of the 16s rDNA sequence of the strain and the Lactobacillus plantarum sequence reaches 99 percent.
Figure GWB0000003175010000092
Combining the biochemical identification result, the strain is identified as Lactobacillus plantarum (Lactobacillus plantarum).
The isolated strain is determined to be Lactobacillus plantarum (Lactobacillus plantarum), named as Lactobacillus plantarum LCZThe microbial inoculum is preserved in the China general microbiological culture Collection center, and the preservation address is as follows: the microbial research institute of China academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, has a preservation number of CGMCC No.16441 and a preservation date of 2018, 9 months and 7 days.
Example 2
This example is used to illustrate the genetic stability study of Lactobacillus plantarum CGMCC NO.16441
In order to examine the acid production and curd performance of CGMCC NO.16441, a test strain is inoculated into a liquid MRS culture medium and cultured for 24-48h at 37 ℃, when the strain recovers the activity, the strain is inoculated into a subculture medium according to the proportion of 4 volume percent (full-cream milk powder is prepared into a reconstituted milk solution of 12 weight percent, and then 6 weight percent of white granulated sugar is added, mixed uniformly and boiled for 10min), and the strain is fermented at 37 ℃. After the milk has solidified, the fermentation time is recorded, and the acidity, texture and the like are measured. The results of the experiment after 10 serial passages are shown in tables 2 and 3.
TABLE 2 CGMCC NO.16441 acid production genetic stability test results
Numbering Fermentation time/h End of fermentation pH After-ripening titration acidity
Generation 10 15 4.53±0.5 89.74±0.32
Generation 1 10 4.32±0.67 95.93±0.78
TABLE 3 CGMCC NO.16441 curd performance genetic stability test results
Numbering Water retention capacity/%) Consistency per gram Viscosity of the oil
Generation 10 48.15±0.48 4721.86±15.80 2388±10.55
Generation 1 42.90±0.37 3886.11±12.47 2852±12.36
As can be seen, after the strain CGMCC NO.16441 is continuously passaged for 10 times, the fermentation end point pH of the fermented milk is 4.53, the after-ripening titration acidity is 89.74, the viscosity reaches 2388, the consistency reaches 4721.86/g, and the acid-producing capability and the curd performance are genetically stable.
Example 3
This example demonstrates the study of the determination of acid productivity of lactic acid bacteria
Inoculating the separated and purified lactobacillus in a closed container filled with MC liquid culture medium after activated microscopic examination, culturing at constant temperature of 37 ℃, taking out 15mI culture solution every 2h, measuring pH value with a precision acidity meter, and measuring for 24 h. The time is used as the abscissa, the pH value of the fermentation culture solution is used as the ordinate, and the pH value dynamic change curve of the acid production of each strain is obtained and is shown in figure 3.
As can be seen, the pH value of the strain CGMCC NO.16441 can reach 3.76 after fermentation for 24 hours, and the acid production capability is higher than that of a commercial strain (the commercial strain No.1 is purchased from Jiangsu Microkang Biotechnology Co., Ltd., lactobacillus strain, the strain type is 1807B, Lactobacillus bulgaricus and Streptococcus thermophilus, and the commercial strain No.1 is purchased from Hebei Yiran Biotechnology Co., Ltd., lactobacillus strain, the strain type is Yo-C698-1F, Lactobacillus delbrueckii subspecies bulgaricus and Streptococcus thermophilus).
Example 4
This example is used to illustrate the study on the production of polysaccharides from Lactobacillus plantarum CGMCC NO.16441
Inoculating the strain into MRS liquid culture medium, culturing at 37 deg.C for 48h, collecting 10mL culture solution, adding 250uL of trichloroacetic acid (with protein removed) with volume fraction of 80%, centrifuging at 8000r/min for 20min in a centrifuge, centrifuging, discarding precipitate, placing supernatant into dialysis bag, and dialyzing with running water for 48 h. Adding 95% ethanol 3 times the volume of the solution after dialysis, refrigerating overnight at 4 deg.C to obtain flocculent precipitate of polysaccharide, centrifuging at 10000r/min for 20min in a centrifuge, discarding supernatant to obtain extracellular polysaccharide, and dissolving the precipitate in 10mL distilled water to obtain extracellular polysaccharide water solution.
Total sugars were determined using the phenol-sulfuric acid method: after 2.0mL of the sample to be detected is added into the test tube, 1mL of 6% phenol and 5mL of concentrated sulfuric acid are sequentially added, the mixture is kept stand for 10min, the mixture is shaken up, the mixture is kept stand for 20min at room temperature, and then the absorbance is detected at 490nm wavelength.
Reducing sugars the 3, 5 dinitrosalicylic acid method was used: adding 3, 5 dinitrosalicylic acid reagent into 0.5mL of sample to be detected, uniformly mixing, carrying out boiling water bath for 5min, cooling, adding 4mL of distilled water, uniformly mixing, and detecting the absorbance at 540 nm.
The polysaccharide content was calculated according to the formula "total sugar-reducing sugar", and the calculation results are shown in table 4 below.
TABLE 4 polysaccharide content of Lactobacillus plantarum CGMCC NO.16441
CGMCC NO. Concentration (mg/L)
Total sugar 803.0±7.5
Reducing sugar 0
Polysaccharides 803.0
Therefore, the screened strain CGMCC NO.16441 has high exopolysaccharide yield of 803.0mg/L and very high polysaccharide production capacity. Is far higher than the similar strains subjected to optimized culture conditions by 150-600 mg/L.
Example 5
This example is used to illustrate the study of Lactobacillus plantarum CGMCC NO.16441 on the simulation of gastric juice and intestinal juice tolerance
Preparing simulated gastric juice: 0.35g pepsin was dissolved in 100mL of 0.2 wt% sterile physiological saline, adjusted to pH3.0 with concentrated HCl, and sterilized through a 0.45um filter.
Preparing simulated intestinal juice: 0.1g trypsin, 1.8g bile salts in sterile solvent 1.1g NaHCO3, 0.2g NaCl and 100mL distilled water, adjusted to pH 8.0 with 0.5M NaOH. The solution was sterilized through a 0.45 μm filter.
Inoculating the activated 3 times bacterial solution into simulated gastric juice (pH3.0) according to the inoculation amount of 10 vol%, mixing, performing anaerobic culture at 37 deg.C, and counting in sampling plates for 0 hr and 3 hr respectively. After 3h of incubation in simulated gastric fluid, 1mL of the culture broth was aspirated and inoculated into 9mL of simulated intestinal fluid (pH8)), anaerobically incubated at 37 ℃ and counted at 24h samples.
Survival (%) (logCFU N)1/logCFU N0)×100％；
Wherein N is1Representing the number of lactic acid bacteria after simulated gastrointestinal culture; n is a radical of 0Representing the number of lactic acid bacteria before the simulated gastrointestinal tract culture.
The results are as follows: the survival rate of CGMCC NO.16441 is 62% after 3h of simulated gastric juice with the pH value of 3, the survival rate is maintained at 39% after 24h of intestinal juice, and the isolated strain shows good gastrointestinal tract tolerance performance.
Example 6
This example illustrates the intestinal adhesion of Lactobacillus plantarum CGMCC NO.16441
The cultured HT-29 cells were digested to prepare a cell suspension (about 10)5one/mL), 2mL was added to a 24-well plate with a cover slip placed thereon, at 37 ℃ with 5% CO2And (4) culturing in 95% air. The culture medium is 1640 culture solution and 10 wt% of inactivated calf serum and L-glutamine. After the cells on the cover glass are basically fully filled, the old cell culture solution is sucked out, the sterile PBS is used for washing for 3 times, and 1mL of bacterial solution (containing 10 thalli) is added into each hole8cfu/mL) and 1mL 1640 cell culture medium at 37 deg.C with 5% CO2And incubating in 95% air for 2 h. The slides were washed with sterile PBS to remove unbound bacteria. Then 10% formaldehyde was fixed for 2h, gram stained, 20 fields were randomly selected under the microscope, the number of adhered bacteria on 100 cells was counted, 3 wells were paralleled for each treatment, and the average number of adhered bacteria per cell was calculated.
The results are as follows: the adhesiveness of CGMCC NO.16441 to HT-29 cells is 1000 bacteria/100 cells, which is higher than 200 bacteria/100 cells of the same strain subjected to optimized culture conditions.
Example 7
This example illustrates the use of Lactobacillus plantarum CGMCC No.16441 in fermented milk
Obtaining raw milk: a qualified standardized raw milk from tribasic group.
Heating raw milk to 60-70 deg.C, adding 6 wt% white sugar, mixing (adding water to initial weight), homogenizing at 65 deg.C and 200bar, and pasteurizing at 85 deg.C for 30 min; cooling to 42 ℃, inoculating by using an inoculation amount of 5 vol%, stirring and uniformly mixing, standing and culturing at 42 ℃ for fixing and forming, and after aging at 4 ℃ for 24 hours, obtaining the fermented milk.
The main roles of lactic acid bacteria in dairy fermentation are acid production and flavor generation. The acid-producing properties of lactic acid bacteria are important characteristics for screening leavening agents for fermented dairy products. Lactic acid bacteria having a strong acid-producing ability generally have a strong tendency to acidify thereafter. Lactic acid bacteria with fast acid production and large acid production amount can bring lower fermentation acidity of final products, which is an effective measure for ensuring low post-acidification. The acid production capacity at the fermentation end point of different strains in the prior art is tested, and the results are shown in table 5. The fermentation end point pH of the detected strain is 4.32 +/-0.67, the after-ripening and after-titration acidity is 95.93 +/-0.78, the result shows that the acidity value of the strain CGMCC No.16441 is increased by 20 percent compared with that of the control lactobacillus, and the fermentation time is obviously shortened.
TABLE 5 comparison table of acid production capacities at fermentation end points of different strains
Strain of bacillus LLD LLL LLC CGMCC NO.16441
Acidity/° T 72.7±2.4 64.4±2.9 71.65±3.4 95.93±0.78
pH value 4.39±0.13 4.77±0.08 4.45±0.14 4.32±0.67
Fermentation time/h 6 6.5 6 5
Note: LLD is lactococcus lactis lactonization variant; LLL is lactococcus lactis subspecies lactis; LLC is lactococcus lactis missible oil subspecies;
the research on flavor substances of lactobacillus plantarum CGMCC NO.16441 fermented milk is implemented by adding 20mL of set yoghurt into 5mL of saturated NaCl solution, shaking and uniformly mixing, transferring 5mL of the solution into a headspace bottle to be tested, and analyzing flavor substance components by GC-MS.
The results of the fermentation flavor test are shown in FIG. 4. In fermented milk, 25 main flavors including ketones, alcohols, esters, ethers, etc. were identified. The common yogurt and the CGMCC No. strain fermented yogurt contain characteristic flavor substances, and also contain flavor components such as 3-hydroxy-2-butanone, 2-heptanone, 2-nonanone and ethyl hexanoate, so that the yogurt is endowed with faint scent flavor.
Example 8
This example illustrates the application of Lactobacillus plantarum CGMCC No.16441 in the preparation of active lactobacillus beverage
Heating purified water to 50-55 deg.C, adding whole milk powder, heating to 65-70 deg.C, homogenizing under 180-200 bar. Cooling, adding Lactobacillus plantarum CGMCC No.16441 as starter in 3-5 vol% of the emulsion, and fermenting at 37-42 deg.C to pH4.0-4.5. Demulsifying, cooling, and storing at low temperature. Heating purified water to 65-75 deg.C, adding mixture of white sugar or other sweetener and stabilizer such as pectin, stirring for 15min, cooling, adding yogurt, adjusting pH to 3.5-4.0, adding essence and water to specified amount, heating to 65-70 deg.C, homogenizing at pressure of 180-200bar and 130 deg.C for 10-20s, cooling, and bottling to obtain lactobacillus beverage, and evaluating its organoleptic properties as shown in Table 6.
TABLE 6 lactic acid bacteria beverage sensory evaluation table
Figure GWB0000003175010000131
The best fermentation strain is selected from different commercial lactobacillus according to the color, taste, smell and tissue state of the lactobacillus fermentation beverage as the scoring standard, and the corresponding sensory evaluation scores selected by the fermentation strain are shown in the following table 7.
TABLE 7 sensory evaluation score
Fermentation strain Color Nourishing and smelling Tissue state Total score
LLD 10 20 12 42
LLL 15 33 20 68
LLC 14 38 20 72
CGMCC No.16441 18 44 22 84
Note: LLD is lactococcus lactis lactonization variant; LLL is lactococcus lactis subspecies lactis; LLC is lactococcus lactis missible oil subspecies;
sensory evaluation is carried out on the lactic acid bacteria beverage fermented by different fermenting agents, so that the experimental result is influenced to a great extent by different fermenting agents under the same fermentation condition, wherein the lactic acid bacteria beverage fermented by CGMCC No.16441 has the fragrance of fermented lactic acid, unique taste, pure white color and uniform consistency.
Example 9
This example illustrates the application of Lactobacillus plantarum CGMCC No.16441 in fermenting cereal beverages
Selecting fresh and full grain raw materials without diseases and insect pests, and soaking the grain raw materials in water according to the range of 1: 2-2: 1 of oat and rice until the grain raw materials completely absorb water and swell.
Taking the soaked raw materials, draining off water, adding water with the weight 8 times of that of the raw materials, pulping and boiling. The slurry enters a colloid mill to circulate for 2-4 min. Adding citric acid to adjust pH of the slurry to 4.0-5.0, adding saccharifying enzyme 1-3% of the composite slurry, and saccharifying at constant temperature of 40-60 deg.C for 4-8 hr.
Cooling saccharified composite pulp, adjusting pH to 6.5-7.5 with sodium bicarbonate, adding Lactobacillus plantarum CGMCC No.16441 as leaven, fermenting and culturing for a period of time to make its concentration reach 108More than cfu/mL, adding white granulated sugar, citric acid, a stabilizer, essence and the like to adjust the flavor, cooling and filling to obtain the product, and evaluating the sense organ of the product, wherein the evaluation standard is shown in Table 8.
TABLE 8 organoleptic evaluation chart of fermented cereal beverages
Figure GWB0000003175010000141
The color, flavor and state of the composite grain fermented beverage are used as scoring standards, the best fermentation strain is selected from different varieties of commercial lactic acid bacteria, and the corresponding sensory evaluation scores selected by the fermentation strains are shown in the following table 9.
TABLE 9 sensory evaluation score
Fermentation strain Color Nourishing and smelling Appearance of the product Total score
LLD 16 20 11 47
LLL 20 33 13 66
LLC 18 27 12 57
CGMCC 26 41 15 82
Note: LLD is lactococcus lactis lactonase variant; LLL is lactococcus lactis subspecies lactis; LLC is lactococcus lactis missible oil subspecies;
sensory evaluation is carried out on the grain fermented beverage fermented by different fermenting agents, so that the different fermenting agents influence the experimental result to a great extent under the same fermentation condition, wherein the grain beverage fermented by the CGMCC No.16441 has grain composite aroma and pleasant fermentation flavor, unique taste, light yellow color and uniform color.
Example 10
The embodiment is used for illustrating the application of lactobacillus plantarum CGMCC No.16441 in the aspect of preparing fermented soybean milk
The fermented soybean milk described in this example was prepared by the following method: soaking soybean with soft water at 75-85 deg.C for 1-2 hr at volume ratio of soft water to soybean of 2: 4, removing soybean hull, removing soaking water, grinding with boiling water at weight ratio of soybean to boiling water of 1: 6-10, maintaining the pulp at 80-85 deg.C for 10-15min, and filtering with 150 mesh screen to obtain soybean milk; adding sucrose 5-10% of the weight of the soybean milk, homogenizing under 15-25Mpa, sterilizing at 95 deg.C for 5-10min, cooling to 28-38 deg.C, inoculating 0.04-0.05% (by weight of the soybean milk) of commercial Lactobacillus bulgaricus, Streptococcus thermophilus and CGMCC NO.16441 starter at a ratio of 1: 1, fermenting at 32-42 deg.C for 6-8 hr, and refrigerating at 4 deg.C to obtain the fermented soybean milk.
Compared with fermented soybean milk prepared by using commercial lactobacillus bulgaricus and streptococcus thermophilus (the ratio of the number of bacteria is 1: 1) as fermenting agents without adding lactobacillus plantarum CGMCC NO. 16441. The result shows that the acidity of the fermented soybean milk added with the lactobacillus plantarum CGMCC NO.16441 is 2.3 times of that of the fermented soybean milk not added, so that the fermentation time can be shortened by 1-1.5 hours by using the lactobacillus plantarum CGMCC NO.16441 to be compounded with commercial strains.
Example 11
This example illustrates the application of Lactobacillus plantarum CGMCC No.16441 in the preparation of fermented fruit and vegetable juice
Selecting fruits and vegetables with uniform and plump particles, cleaning dirt on the surface of the fruits and vegetables with clear water, soaking the fruits and vegetables in 0.005-0.01g/kg of sodium hypochlorite solution for 7-10 minutes for disinfection, washing the fruits and vegetables for standby, and cutting the fruits and vegetables to be cut into pieces; scalding with boiling water for 3-5min, and optionally treating with mixed solution of 0.5% citric acid and 0.3% Vc; pulping fruits and vegetables with tissue masher for 3-5 min; adding 0.006-0.01% of pectinase and 0.001% of cellulase, and performing enzymolysis at 35-52 deg.C for 1-2 hr; filtering with a double-layer 80-mesh filter screen to obtain clear liquid; sterilizing with microwave for 2min or 80 deg.C for 15min to inactivate enzyme; selecting Lactobacillus plantarum CGMCC NO.16441 from inclined planeCulturing in broth for 24 h; selecting the strain, culturing in a small amount of fruit and vegetable juice for 20 hr, inoculating the fermented seed liquid into the fruit and vegetable juice, culturing at appropriate temperature for 24 hr until pH reaches 3.8-4.1 and lactobacillus amount reaches (10)8-109) cfu/mL. Compared with the fermented fruit and vegetable juice using lactobacillus casei as a leaven, the fermented fruit and vegetable juice of lactobacillus plantarum has smaller acidity change value during refrigeration, smaller post-acidification and good fermentation stability.
Example 12
The embodiment is used for illustrating the application of lactobacillus plantarum CGMCC No.16441 in the aspect of preparing fermented feed
Drying crop straws until the water content is less than 15 wt%, and crushing; harvesting herbaceous plants, drying until the water content is less than 15%, and preparing into chopped grass; premixing rice bran, wheat bran, salt, lactobacillus plantarum CGMCC NO.16441 direct vat set starter and cellulase, and then gradually expanding and uniformly mixing with crop straws and herbal plant raw materials; compacting, bundling and film sealing the uniformly mixed micro-storage feed to prepare the micro-storage feed with the specification of 30-100 kg/package; naturally fermenting the obtained product for 12-18 days. The crop straw is one or more selected from vegetable straw, peanut vine, corn stalk and the like. The herbaceous plant is one or more selected from common Astragalus sinicus herbaceous plants.
Compared with the feed fermented by the commercial compound probiotics, the lactic acid content in the feed fermented by the lactobacillus plantarum CGMCC NO.16441 is 21.28g/kg, which is 4.4 times of the lactic acid content in the commercial compound probiotics fermented feed.
Example 13
This example illustrates the application of Lactobacillus plantarum CGMCC NO.16441 in the production of viable bacteria preparation
The screened lactobacillus plantarum is prepared into a viable bacteria preparation. Inoculating Lactobacillus plantarum CGMCC NO.16441 into MRS liquid culture medium according to the inoculation amount of 2-4% (v/v), and culturing at 37-42 deg.C for 20-24h to make the viable count of Lactobacillus plantarum CGMCC NO. reach 10 8centrifuging at above cfu/mL (4000rpm, 10min), washing precipitate with PBS buffer solution of pH6.5-7.5 twice, adding skimmed milk and trehalose as lyophilized protectant, and concoctingWhole cell concentration to 109cfu/mL, and after being uniformly mixed, the mixture is subjected to vacuum freeze drying to obtain the viable bacteria preparation. The live bacteria preparation contains viable bacteria number of 108-1010cfu/mL, the microbial inoculum can be directly added into a fermentation product for fermentation, or can be added with proper auxiliary materials according to a conventional method to prepare various formulations which are acceptable in food industry or clinic, such as capsules, microcapsules, tablets, powder and the like.
The viable bacteria preparation prepared from the screened lactobacillus plantarum also comprises a product for keeping the activity of the strains by technical means such as domestication and the like.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including various technical features being combined in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
SEQUENCE LISTING
<110> Zhongliang Nutrition and health research institute Co., Ltd
<120> lactobacillus plantarum strain and application thereof
<130> P55059COF-55058
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1185
<212> DNA
Nucleotide sequence of 16s rDNA gene of <213> CGMCC NO.16441
<400> 1
gggggcctaa tacatgccaa gtcgaacgaa ctctggtatt gattggtgct tgcatcatga 60
tttacatttg agtgagtggc gaactggtga gtaacacgtg ggaaacctgc ccagaagcgg 120
gggataacac ctggaaacag atgctaatac cgcataacaa cttggaccgc atggtccgag 180
tttgaaagat ggcttcggct atcacttttg gatggtcccg cggcgtatta gctagatggt 240
ggggtaacgg ctcaccatgg caatgatacg tagccgacct gagagggtaa tcggccacat 300
tgggactgag acacggccca aactcctacg ggaggcagca gtagggaatc ttccacaatg 360
gacgaaagtc tgatggagca acgccgcgtg agtgaagaag ggtttcggct cgtaaaactc 420
tgttgttaaa gaagaacata tctgagagta actgttcagg tattgacggt atttaaccag 480
aaagccacgg ctaactacgt gccagcagcc gcggtaatac gtaggtggca agcgttgtcc 540
ggatttattg ggcgtaaagc gagcgcaggc ggttttttaa gtctgatgtg aaagccttcg 600
gctcaaccga agaagtgcat cggaaactgg gaaacttgag tgcagaagag gacagtggaa 660
ctccatgtgt agcggtgaaa tgcgtagata tatggaagaa caccagtggc gaaggcggct 720
gtctggtctg taactgacgc tgaggctcga aagtatgggt agcaaacagg attagatacc 780
ctggtagtcc ataccgtaaa cgatgaatgc taagtgttgg agggtttccg cccttcagtg 840
ctgcagctaa cgcattaagc attccgcctg gggagtacgg ccgcaaggct gaaactcaaa 900
ggaattgacg ggggcccgca caagcggtgg agcatgtggt ttaattcgaa gctacgcgaa 960
gaaccttacc aggtcttgac atactatgca aatctaagag attagacgtt cccttcgggg 1020
acatggatac aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt tgggttaagt 1080
cccgcaacga gcgcaaccct tattatcagt tgccagcatt aagttgggca ctctggggga 1140
gactgccggt gacaaaccgg aagaaagggg ggggatgaag ttcaa 1185

Claims (10)

1. The Lactobacillus plantarum is characterized in that the Lactobacillus plantarum is classified and named as Lactobacillus plantarum and is preserved in China general microbiological culture Collection center (CGMCC), and the preservation number of the Lactobacillus plantarum is CGMCC No. 16441.
2. Use of the lactobacillus plantarum of claim 1 in the preparation of fermented dairy products, fermented cereal beverages, fermented soy products, fermented fruit delicacies, fermented feed.
3. Use according to claim 2, wherein the use comprises the step of contacting the lactobacillus plantarum strain of claim 1 with an adapted fermentation substrate.
4. Use of lactobacillus plantarum as defined in claim 1 for the preparation of a ready-to-use starter culture.
5. A direct vat set starter prepared by fermenting the Lactobacillus plantarum of claim 1 in a fermentation medium.
6. A method of preparing the ready-to-use starter culture of claim 5, comprising:
(1) fermenting and culturing the lactobacillus plantarum of claim 1 in a fermentation medium to reach a viable count of 10 8More than cfu/mL;
(2) centrifuging the zymocyte liquid obtained in the step (1), washing with a buffer solution, adding a freeze-drying protective agent, and adjusting the concentration of viable bacteria to 1010More than cfu/mL, and after being uniformly mixed, the mixture is subjected to vacuum freeze drying to obtain the direct vat set starter.
7. Use of the ready-to-use starter culture according to claim 5 and/or the ready-to-use starter culture prepared by the method according to claim 6 in the field of preparing fermented dairy products, fermented cereal beverages, fermented soy products, fermented fruit cooked products and fermented feed.
8. Use of lactobacillus plantarum as claimed in claim 1 for the preparation of a live bacterial formulation.
9. The use according to claim 8, wherein the viable bacteria formulation comprises capsules, tablets, oral liquid and powders.
10. A live bacterial preparation, which is prepared by fermenting and culturing the Lactobacillus plantarum of claim 1 in a fermentation medium.
CN201880030861.0A 2018-11-12 2018-12-27 Fermented lactobacillus plantarum and application thereof Active CN111436203B (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CN2018113415222 2018-11-12
CN201811341522 2018-11-12
PCT/CN2018/124327 WO2020098097A1 (en) 2018-11-12 2018-12-27 Fermented lactobacillus plantarum and application thereof

Publications (2)

Publication Number Publication Date
CN111436203A CN111436203A (en) 2020-07-21
CN111436203B true CN111436203B (en) 2022-06-28

Family

ID=70731942

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201880030861.0A Active CN111436203B (en) 2018-11-12 2018-12-27 Fermented lactobacillus plantarum and application thereof

Country Status (3)

Country Link
CN (1) CN111436203B (en)
AU (1) AU2018423072B2 (en)
WO (1) WO2020098097A1 (en)

Families Citing this family (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110885768B (en) * 2019-10-11 2022-07-19 东莞理工学院 Staphylococcus saprophyticus and application thereof in preparation of dry fermented sausage
CN111778180B (en) * 2020-06-12 2021-12-14 北京三元食品股份有限公司 Breast milk source lactobacillus plantarum and application thereof
CN111657324A (en) * 2020-07-17 2020-09-15 上海应用技术大学 Natural apple fermentation liquid coarse cereal egg-free cake and preparation method thereof
CN112205327A (en) * 2020-10-28 2021-01-12 广东梁氏水产种业有限公司 Application of lactobacillus plantarum in improving domestication rate of artificial feed for siniperca chuatsi
CN112553124B (en) * 2020-12-28 2022-06-07 福建省农业科学院农业工程技术研究所 Lactobacillus plantarum strain and application thereof
CN114806918A (en) * 2020-12-31 2022-07-29 陕西师范大学 Lactobacillus plantarum NUFF0528 and application thereof
CN112708652B (en) * 2021-01-11 2022-10-21 青蛙王子（福建）婴童护理用品有限公司 Avocado oil-in-water liquid state fermentation product and preparation method and application thereof
CN112813006B (en) * 2021-02-08 2023-08-18 广东石油化工学院 Lactobacillus plantarum and application thereof in fermentation of fruit and vegetable juice
CN113068782B (en) * 2021-04-01 2023-03-28 新疆农业科学院微生物应用研究所(中国新疆—亚美尼亚生物工程研究开发中心) Brown active lactobacillus beverage and preparation method thereof
CN112877266B (en) * 2021-04-16 2022-11-18 四川师范大学 Lactobacillus plantarum QS7T and application thereof
CN113234639B (en) * 2021-06-16 2022-08-30 广东海天创新技术有限公司 Lactobacillus plantarum ZF632 and application thereof
CN113717879B (en) * 2021-07-22 2023-04-14 广东海天创新技术有限公司 Lactobacillus plantarum ZF603 and application thereof
CN113773989B (en) * 2021-08-30 2023-06-13 大连工业大学 Application of extracellular polysaccharide produced by lactobacillus plantarum Y12 in relieving dysentery caused by shigella flexneri
CN113980847B (en) * 2021-09-02 2023-07-28 中粮营养健康研究院有限公司 Lactobacillus plantarum for improving taste of noodles, starter and application thereof, and method for preparing flour product
CN114196566B (en) * 2021-10-20 2023-08-29 君乐宝乳业集团有限公司 Streptococcus thermophilus JMCC0033 and application thereof
CN114350536B (en) * 2021-10-20 2023-06-06 中粮营养健康研究院有限公司 Lactobacillus plantarum, microbial inoculum and preparation method thereof, application of lactobacillus plantarum and microbial inoculum, fermented food and preparation method of fermented food
CN114276968B (en) * 2022-01-28 2023-11-10 河北经贸大学 Lactobacillus plantarum Z43 and application thereof in low-salt fermented sausage
CN114958652B (en) * 2022-04-26 2024-04-19 大连工业大学 Compound starter containing lactobacillus plantarum Y44 and fermented compound milk thereof
CN114940956B (en) * 2022-05-10 2023-04-21 中国农业科学院农产品加工研究所 Lactobacillus plantarum WL-100 and application thereof
CN115011533A (en) * 2022-08-09 2022-09-06 中国肉类食品综合研究中心 Soybean milk leavening agent and application thereof in low-sugar solidified fermented soybean milk
CN115500455B (en) * 2022-09-30 2024-03-26 江南大学 Method for improving total phenol content of oat whole grains through physical synergistic biological treatment
CN116218710B (en) * 2022-12-21 2023-12-12 天津博菲德科技有限公司 Lactobacillus casei and method for preparing high dietary fiber by fermenting vegetable tails by using same
CN115886169A (en) * 2022-12-21 2023-04-04 天津市工业微生物研究所有限公司 Black rice clear juice fermented beverage and preparation process thereof
CN115927117A (en) * 2023-01-04 2023-04-07 兰州大学 Lactobacillus plantarum and application thereof
CN116555114B (en) * 2023-05-17 2024-06-18 内蒙古农业大学 Ferment and application thereof, milk bean curd and preparation method thereof
CN117070425B (en) * 2023-10-10 2024-03-08 广东益可维生物技术有限公司 Technological method for improving metabolic stability of probiotics in organism and probiotics freeze-dried powder
CN117535194A (en) * 2023-11-16 2024-02-09 黑龙江禾家福农业科技有限公司 Lactobacillus plantarum

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102586155A (en) * 2012-03-07 2012-07-18 江南大学 Lactobacillus plantarum N13 and use thereof
CN104498402A (en) * 2014-12-19 2015-04-08 北京工商大学 Physiologically active lactobacillus plantarum for producing exopolysaccharide and application thereof
CN105494629A (en) * 2015-11-12 2016-04-20 南昌大学 Preparation method of yoghourt
CN108165512A (en) * 2018-02-05 2018-06-15 富乐顿生物工程科技（北京）有限公司 A kind of extracellular polysaccharide spatial plant lactobacillus SS18-119 and its application in biological antioxidant activity is improved

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101864375B (en) * 2010-01-15 2011-11-02 吉林省农业科学院 Lactobacillu plantarurn with function of reducing cholesterol and application thereof
CN105176875A (en) * 2015-10-10 2015-12-23 塔里木大学 Lactobacillus plantarum and application thereof
CN105519667A (en) * 2015-11-12 2016-04-27 南昌大学 Yogurt starter culture
CN105494630A (en) * 2015-11-12 2016-04-20 南昌大学 Preparation method of carrot juice yoghourt beverage
CN105494638A (en) * 2015-11-12 2016-04-20 南昌大学 Preparation method of kiwi fruit granule yoghourt beverage
CN105505813A (en) * 2015-11-12 2016-04-20 南昌大学 Strongly-bacteriostatic lactobacillus plantarum
CN105494639A (en) * 2015-11-12 2016-04-20 南昌大学 Preparation method of mango granule yoghourt beverage
CN105685220A (en) * 2015-11-12 2016-06-22 南昌大学 Preparation technology of probiotic flavored yogurt
CN105494640A (en) * 2015-11-12 2016-04-20 南昌大学 Preparation method of strawberry granule yoghourt beverage
KR101800174B1 (en) * 2016-02-22 2017-11-22 롯데칠성음료주식회사 Lactobacillus plantarum LRCC5308 strain having improved acid-resistance, method for enhancing acid-resistant stability, fruits and vegetables beverage containing the strain, and method for manufacturing thereof
CN105821093A (en) * 2016-04-05 2016-08-03 南昌大学 Lactobacillus plantarum exopolysaccharide and preparation method thereof
CN107227277A (en) * 2017-07-17 2017-10-03 浙江鸣食品股份有限公司 A kind of Lactobacillus plantarum E680 and its application

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102586155A (en) * 2012-03-07 2012-07-18 江南大学 Lactobacillus plantarum N13 and use thereof
CN104498402A (en) * 2014-12-19 2015-04-08 北京工商大学 Physiologically active lactobacillus plantarum for producing exopolysaccharide and application thereof
CN105494629A (en) * 2015-11-12 2016-04-20 南昌大学 Preparation method of yoghourt
CN108165512A (en) * 2018-02-05 2018-06-15 富乐顿生物工程科技（北京）有限公司 A kind of extracellular polysaccharide spatial plant lactobacillus SS18-119 and its application in biological antioxidant activity is improved

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
产胞外多糖植物乳杆菌C88发酵条件优化;张岩春等;《中国乳品工业》;20100725(第07期);18-21页 *
植物乳杆菌SC9直投式发酵剂的研究;周洋等;《农产品加工(学刊)》;20131120(第21期);11-14页 *
植物乳杆菌YW11生产胞外多糖的发酵条件研究;曹永强 等;《食品科学技术学报》;20160119;第34卷(第1期);42-49页，78页 *

Also Published As

Publication number Publication date
AU2018423072A1 (en) 2020-05-28
AU2018423072B2 (en) 2021-05-13
CN111436203A (en) 2020-07-21
WO2020098097A1 (en) 2020-05-22

Similar Documents

Publication Publication Date Title
CN111436203B (en) Fermented lactobacillus plantarum and application thereof
US4702923A (en) Lyophilized kefir yoghurt health food
CN105533544A (en) Natural fruit and vegetable enzyme product
CN102318806B (en) Preparation method of probiotics fermented pumpkin and carrot vegetable powder
CN112195139B (en) Lactobacillus plantarum strain LDVS007 and application thereof
CN107198076A (en) A kind of bacillus coagulans probiotics normal temperature viable bacteria beverage and preparation method thereof
CN113061543A (en) Lactobacillus plantarum and application thereof
CN107227278A (en) A kind of Lactobacillus plantarum A11 and its application
CN110301565B (en) Fermented fruit and vegetable juice and preparation method thereof
KR20170045050A (en) Cereals fermented by lactic acid bacteria and method for manufacturing the same
CN114231473A (en) Probiotic lactobacillus plantarum and application thereof in preparation of low-salt fermented meat food
CN111117926B (en) Strain for high yield of gamma-aminobutyric acid and application of strain to preparation of gamma-aminobutyric acid-rich fruit juice
CN109294940A (en) The purposes of corn lactobacillus mutagenic bacteria and high-yield lactic acid
CN114231381B (en) Preparation method of mixed bacteria compound fermentation roxburgh rose fruit vinegar
CN113249236B (en) Saccharomyces cerevisiae, leavening agent, preparation method of saccharomyces cerevisiae and leavening agent, application of saccharomyces cerevisiae and leavening agent in preparation of fermented food, fermented rice cake and preparation method of fermented rice cake
JP6955808B1 (en) How to make fermented honey
CN109337833B (en) Biological preparation of mutagenic leuconostoc mesenteroides and application thereof
CN101144064B (en) Lactobacillus brevis with anti-mutagenesis active and capable of producing exopolysaccharide and use thereof
CN105018377B (en) A kind of Lactobacillus casei of high yield ethyl butyrate and its application
CN114304618A (en) Method for enriching active ingredients of dandelion roots by fermentation and application of method
CN112877266A (en) Lactobacillus plantarum QS7T and application thereof
Maduka et al. Fermentation of tigernut by lactic acid bacteria and tigernut-milk drink fermentation by lactic acid bacteria as a potential probiotic product
CN114350536B (en) Lactobacillus plantarum, microbial inoculum and preparation method thereof, application of lactobacillus plantarum and microbial inoculum, fermented food and preparation method of fermented food
CN113913342B (en) Leuconostoc mesenteroides and application thereof
CN115644332B (en) Rosa roxburghii and coix seed composite beverage and preparation method thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant