CN117987311A - Lactobacillus paracasei YYS-EN2 for producing cellulase, pectase, tannase and phytase and application thereof - Google Patents
Lactobacillus paracasei YYS-EN2 for producing cellulase, pectase, tannase and phytase and application thereof Download PDFInfo
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- CN117987311A CN117987311A CN202410139976.0A CN202410139976A CN117987311A CN 117987311 A CN117987311 A CN 117987311A CN 202410139976 A CN202410139976 A CN 202410139976A CN 117987311 A CN117987311 A CN 117987311A
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- China
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- yys
- lactobacillus paracasei
- fermented
- powder
- tannase
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- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 claims abstract description 20
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus paracasei YYS-EN2 for producing cellulase, pectase, tannase and phytase and application thereof. The Lactobacillus paracasei (Lactobacillus paracasei) YYS-EN2 is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.28531. The lactobacillus paracasei YYS-EN2 provided by the invention has the capability of producing cellulase, pectase, tannase and phytase, can degrade plant cellulose and pectin, decompose tannin and phytic acid, can reduce bitter taste and improve clarity of products when being applied to fermentation products, can be widely applied to production of fermentation products such as fermented traditional Chinese medicines, fermented plant-derived nutrition powder, fermented plant-derived polypeptide powder and the like, can also be applied to probiotic products and functional foods, and has the effects of improving flora structure and promoting digestion and absorption when being subjected to intestinal field planting in human bodies.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to lactobacillus paracasei YYS-EN2 for producing cellulase, pectase, tannase and phytase and application thereof.
Background
Crude fiber materials such as cellulose, pectin, tannin and the like are abundant in traditional Chinese medicines, grains, bran, fruits and vegetables, are not easy to hydrolyze, and the crystalline rigid structure of the crude fiber materials causes great production difficulty; in the product processing process, the processing steps of the product can be increased, and the yield is reduced; even to some extent damaging the nutritional value of the reduced product. The physical and chemical degradation conditions are extreme, the period is long, and secondary pollution is easy to bring; therefore, the microbial degradation method becomes the most effective, healthy and environment-friendly method. The cellulase and the pectase can hydrolyze crude plant fibers, are favorable for dissolving and releasing substances such as starch, protein and the like, are favorable for nutrition absorption, improve the value of the product, and are widely applied to extraction and clarification of traditional Chinese medicines, fruits and vegetables and fermentation of tea and coffee. Tannase and phytase can degrade tannins and phytic acid, and can improve the squeezing and color of fruit juice and vegetables; enhancing the flavor and color of wine.
In conclusion, the cellulase, the pectase, the tannase and the phytase have wider application prospects in fermentation of traditional Chinese medicines, plant-based fermented beverages, fermentation of plant-derived nutrition powder, fermentation of plant-derived polypeptide powder and fermentation of plant-derived protein powder.
The Chinese patent of application No. CN202110864302.3 and publication No. 20230203 discloses a Streptomyces strain GZUI RF-Y1, a method for preparing cellulase fermentation broth and application thereof, wherein the cellulase activity in the fermentation broth of the strain reaches 111.0 U.mL -1. In this technique, the strain degrading the cellulose function is a mold.
The invention patent of China with the application number of CN202111637611.3 and the publication date of 20220412 discloses an aspergillus oryzae strain with the cellulase activity of 18.6U.g -1 and the acid pectase activity of 33.1U.g -1 in high-salt and high-nitrogen fermented foods. In this technique, the strain degrading the cellulose function is a mold.
The mould can produce cellulase and/or pectase, and does not have the functions of producing cellulase, pectase, tannase and phytase. Also, at present, most of bacterial strains degrading cellulose, pectin, tannin and phytic acid are mould or bacillus, and mould or bacillus are less beneficial bacteria and cannot be applied to food fermentation.
In summary, how to develop natural probiotics with cellulase, pectase, tannase and phytase production is the technical problem in the field to be solved.
Disclosure of Invention
In order to solve the defects of the prior art mentioned in the background art, the invention provides lactobacillus paracasei YYS-EN2, wherein the lactobacillus paracasei YYS-EN2 is a natural strain, has the capability of producing cellulase, pectase, tannase and phytase, can degrade plant cellulose and pectin, decompose tannin and phytic acid, can reduce bitter taste and improve the clarity of products, and is widely applied to the production of fermented products such as fermented traditional Chinese medicines, fermented plant-derived nutrition powder, fermented plant-derived polypeptide powder and the like.
The lactobacillus paracasei YYS-EN2 provided by the invention, latin Wen Xueming: lactobacillus paracasei, and is preserved in China general microbiological culture Collection center (CGMCC) at 25/09/2023 with the preservation number of CGMCC No.28531.
The Lactobacillus paracasei YYS-EN2 is isolated from saliva of the oral cavity of a healthy adult, and is subjected to sequencing analysis, and is aligned to be highly homologous with the Lactobacillus paracasei through Blast sequences, and is named as Lactobacillus paracasei YYS-EN2.
Among them, lactobacillus paracasei YYS-EN2 has the ability to produce cellulase, pectinase, tannase and phytase, and has the ability to degrade cellulose, pectin, tannin and phytic acid.
The invention also provides an application of lactobacillus paracasei YYS-EN2 in preparing a fermentation product. The lactobacillus paracasei YYS-EN2 is applied to the preparation of a fermentation product, and can obviously reduce the content of cellulose and pectin in the product.
In some embodiments, the fermentation product includes, but is not limited to, fermented traditional Chinese medicine, fermented nutritional powder, fermented polypeptide powder, fermented protein powder, ferment, silage.
In some embodiments, the fermented nutritional powder comprises a fermented plant-derived nutritional powder; the fermented polypeptide powder comprises fermented plant-derived polypeptide powder; the fermented protein powder comprises fermented plant-derived protein powder.
In some embodiments, the fermented plant-derived nutritional powder comprises a fermented corn nutritional powder. Lactobacillus paracasei YYS-EN2 is applied to a corn nutrition powder product, and can ferment corn husks and corn embryos, decompose cellulose and pectin, reduce tannin and phytic acid, and strengthen the flavor and taste of the corn nutrition powder.
The invention also provides a method for producing cellulase, pectase, tannase and phytase: inoculating lactobacillus paracasei YYS-EN2 into an enzyme-producing liquid culture medium, culturing at the temperature of between 30 and 40 ℃ for 48 to 72 hours, and carrying out solid-liquid separation on the enzyme-producing liquid culture medium after fermentation culture to obtain a supernatant; the supernatant is an enzyme solution containing cellulase and/or pectase and/or tannase and/or phytase.
The invention also provides a freeze-dried product, the components of which comprise lactobacillus paracasei YYS-EN2 as described above.
In some embodiments, the viable count of lactobacillus paracasei YYS-EN2 within the lyophilized product is (2-8) ×10 1CFU·g-1.
The invention also provides a preparation method of the freeze-dried product, which comprises the following preparation steps:
1) Preparing lactobacillus paracasei YYS-EN2 seed liquid;
2) Performing seed liquid expansion culture;
3) Fermenting and culturing the seed liquid to obtain fermentation liquid;
4) Centrifuging, and discarding supernatant to obtain bacterial sludge;
5) Uniformly mixing bacterial mud and a freeze-drying protective agent, and then emulsifying and embedding to obtain emulsion;
6) Freeze-drying the emulsion, and pulverizing to obtain freeze-dried powder of Lactobacillus paracasei YYS-EN 2.
The invention also provides a microbial inoculum, the components of which comprise the freeze-dried powder and/or the components of which comprise lactobacillus paracasei YYS-EN2.
In some embodiments, the microbial agent further comprises other ingredients that are currently available for microbial agents, such as at least one of prebiotics, fillers, acidulants, solvents, solubilizing agents, co-solvents, emulsifiers, colorants, binders, disintegrants, lubricants, wetting agents, stabilizers, glidants, flavoring agents, preservatives, coating materials, fragrances, anti-binding agents, integration agents, thickening agents, and inclusion agents.
The invention also provides application of lactobacillus paracasei YYS-EN2 in preparation of a probiotic product.
The invention also provides the use of lactobacillus paracasei YYS-EN2 as described above for the preparation of a functional product comprising at least one of the following effects:
(1) Has cellulose decomposing ability;
(2) Has pectin decomposing ability;
(3) Has the ability of decomposing tannin;
(4) Has the capability of decomposing phytic acid.
In some embodiments, the functional product comprises a food, a nutraceutical; lactobacillus paracasei YYS-EN2 can be used for preparing food or health product.
Based on the characteristics, the lactobacillus paracasei YYS-EN2 provided by the invention has the following beneficial effects:
the lactobacillus paracasei YYS-EN2 provided by the invention has the capability of producing cellulase, pectase, tannase and phytase, and can degrade plant cellulose, pectin, tannin and phytic acid.
The lactobacillus paracasei YYS-EN2 provided by the invention has the capability of producing cellulase, can degrade cellulose, and can be applied to development of fermented traditional Chinese medicines, fermented plant-derived nutrition powder and silage products.
The pectin has the pectase degradation capability, can be widely applied to the production of products such as fermented traditional Chinese medicines, fruit juice, vegetables, wines, enzymes and the like, can obviously reduce the pectin content in raw materials, and improves the clarity and the yield of the products.
The product has the capability of producing the tannase, can be widely applied to the production of products such as traditional Chinese medicines, wines, enzymes and the like, can reduce the content of the tannase in the wine, and improves the mouthfeel of the products.
The phytase-producing enzyme has the capability of producing phytase, can be widely applied to flour and soybean food processing, improves the degradation rate of phytic acid, and improves the nutrition and commodity value of the phytic acid.
In conclusion, the lactobacillus paracasei YYS-EN2 provided by the invention has the functions of producing cellulase, pectase, tannase and phytase, can be applied to various probiotic fermentation products, and lays a foundation for providing green healthy, high-nutrition-value and high-utilization-rate plant-source crop products.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the drawings that are needed in the embodiments or the description of the prior art will be briefly described below, it will be obvious that the drawings in the following description are some embodiments of the present invention, and that other drawings can be obtained according to these drawings without inventive effort to a person skilled in the art.
FIG. 1 is a graph of the colony of Lactobacillus paracasei YYS-EN2 on MRS plate medium;
FIG. 2 is a graph showing a glucose standard curve;
FIG. 3 is a graph showing a D-galacturonic acid standard curve;
FIG. 4 is a gallic acid standard curve display diagram;
FIG. 5 is a graph showing a standard curve of sodium phytate.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The scheme of the invention is as follows:
The invention provides lactobacillus paracasei (Lactobacillus paracasei) YYS-EN2:
Lactobacillus paracasei (Lactobacillus paracasei) YYS-EN2 was deposited with the China general microbiological culture Collection center, with accession number No. 3, the west way 1, north Star, the korean region, of beijing, at year 2023, and month 09, 25: CGMCC No.28531;
The source is as follows: the lactobacillus paracasei is isolated from saliva of oral cavity of a healthy adult, and the strain is subjected to sequencing analysis, and is highly homologous to the lactobacillus paracasei through Blast sequence alignment, and is named as lactobacillus paracasei YYS-EN2.
Colony morphology: colonies in MRS solid culture medium are milky white, round and smooth and neat in edge.
The functions are as follows: has the capability of producing cellulase and pectase, can degrade cellulose and pectin, has the capability of producing tannase and phytase, and can decompose tannin and phytic acid.
The invention also provides an operation example of a method for producing cellulase and/or pectinase and/or tannase and/or phytase:
Inoculating lactobacillus paracasei YYS-EN2 into an enzyme-producing liquid culture medium, culturing at the temperature of between 30 and 40 ℃ for 48 to 72 hours, and carrying out solid-liquid separation on the enzyme-producing liquid culture medium after fermentation culture to obtain a supernatant; the supernatant is an enzyme solution containing cellulase and/or pectase and/or tannase and/or phytase.
The invention also provides a preparation method of the freeze-dried product, which comprises the following preparation steps:
1) Preparing lactobacillus paracasei YYS-EN2 seed liquid;
2) Performing seed liquid expansion culture;
3) Fermenting and culturing the seed liquid to obtain fermentation liquid;
4) Centrifuging the fermentation liquor to obtain bacterial sludge;
5) Mixing bacterial mud with a freeze-drying protective agent, and then emulsifying and embedding to obtain emulsion;
6) Freeze-drying the emulsion, and pulverizing to obtain lactobacillus paracasei YYS-EN2 freeze-dried powder.
The invention also provides an operation example of an application method in the preparation of the corn nutrition powder:
Crushing corn embryo and corn husk, treating with lipase and amylase, sterilizing at high temperature, adding lactobacillus paracasei YYS-EN2 bacterial powder and/or lyophilized powder, mixing, fermenting, sterilizing at high temperature, and drying.
The technical solutions provided by the present invention are described in detail below in conjunction with examples for further illustrating the present invention, but they should not be construed as limiting the scope of the present invention.
EXAMPLE 1 isolation and characterization of Lactobacillus paracasei YYS-EN2
Separating:
Sterile sampling from oral saliva of a healthy adult, dipping the oral saliva sample of the healthy adult into a sterile homogenizing bag by using a sterile cotton swab, adding 45mL of 0.9% physiological saline, and homogenizing and uniformly mixing to obtain a sample. 100. Mu.L of the sample was pipetted for 10-fold serial gradient dilutions, 100. Mu.L of the sample with dilution factor of 10 -2、10-3、10-4 was pipetted respectively, and the samples were spread on MRS solid plate medium containing 2.5% CaCO 3 and incubated for 24h at 37℃in an inverted position. Selecting colony with good growth vigor and large calcium dissolving ring, repeatedly separating and purifying by plate streak separation method until single colony is obtained, screening the separated bacteria to obtain high-yield cellulase, pectase, tannase and phytase, named YYS-EN2, and adding glycerol for preservation in a fungus bank at-80deg.C.
The colony morphology of the isolated and purified Lactobacillus paracasei YYS-EN2 is as follows: colonies in MRS solid culture medium are milky white, round and smooth and neat in edge.
Wherein, the bacteria for obtaining the high-yield cellulase, pectase, tannase and phytase are obtained by screening, and the process comprises the following steps:
screening cellulase-producing strains:
1 screening Medium (g.L-1): 20g of sodium carboxymethylcellulose (CMC-Na), 10g of peptone, 5g of yeast powder, 5g of sodium chloride, 1g of monopotassium phosphate and 0.3g of magnesium sulfate; congo red 0.4g;
2 screening of strains: inoculating bacteria into MRS liquid culture medium from-80deg.C refrigerator, and culturing at 37deg.C for 24 hr; then 100 mu L of culture solution is placed on a screening culture medium plate for culturing for 48 hours at 37 ℃ by a oxford cup method, the diameter of a transparent circle is observed, a strain with high-yield cellulase is selected according to the diameter of the transparent circle, and the enzyme activity of the cellulase is measured, so that the strain with high-yield cellulase is screened.
Similarly, bacterial screening of high-yield pectase, tannase and phytase is sequentially carried out, and the process is specifically as follows:
Screening pectase-producing strains:
1. screening media (g.L-1): 10g of pectin, 0.2g of Congo red, 1.5g of peptone, 4g of dipotassium hydrogen phosphate, 2g of monopotassium phosphate, 4g of ammonium sulfate and 0.5g of magnesium sulfate heptahydrate.
2 Screening of strains: inoculating bacteria into MRS liquid culture medium from-80deg.C refrigerator, and culturing at 37deg.C for 24 hr; then 100 mu L of culture solution is placed on a screening culture medium plate for culturing for 48 hours at 37 ℃ by a oxford cup method, the diameter of a transparent circle is observed, strains with high pectase yield are selected according to the diameter of the transparent circle, enzyme activity determination of pectase is carried out, and bacteria with high pectase yield are screened out.
Screening of tannase-producing Strain
1. Screening media (g.L-1): 3g of sodium nitrate, 1g of dipotassium hydrogen phosphate, 0.5g of potassium chloride and 0.5g of magnesium sulfate; 0.04g of bromophenol blue and 10g of tannic acid, and sterilizing separately.
2 Screening of strains: inoculating bacteria into MRS liquid culture medium from-80deg.C refrigerator, and culturing at 37deg.C for 24 hr; then 100 mu L of culture solution is placed on a screening culture medium flat plate for culturing for 48 hours at 37 ℃, tannic acid and bromophenol blue indicator are added into the screening culture medium, if the strain produces the tannin enzyme, tannic acid in the culture medium is hydrolyzed to produce gallic acid, and the bromophenol blue indicator around the colony is changed from blue-purple to yellow, so that an obvious color-changing ring is formed around the colony. And selecting a strain with high tannase yield according to the color depth and the diameter of the color-changing ring, and carrying out tannase activity measurement to screen out the strain with high tannase yield.
Phytase producing strain selection
1. Screening medium (g.L -1): 15g of glucose, 5.0g of ammonium nitrate, 0.5g of potassium chloride, 0.5g of magnesium sulfate, 0.3g of ferrous sulfate, 0.3g of manganese sulfate and 2g of calcium phytate.
2. Strain screening: inoculating bacteria into MRS liquid culture medium from-80 refrigerator, and culturing at 37deg.C for 24 hr; then 100 mu L of culture solution is placed on a screening culture medium plate for culturing for 48 hours at 37 ℃ by adopting an oxford cup method, and the strain producing phytase can hydrolyze calcium phytate to form transparent rings around bacterial colonies. And selecting a strain with high phytase yield according to the diameter of the transparent ring, measuring the enzyme activity of the phytase, and screening out the strain with high phytase yield.
The formula of the MRS liquid culture medium is as follows: 10.0g of beef extract, 20.0g of glucose, 10.0g of tryptone, 5.0g of yeast extract powder, 1.0mL of Tween 80, 2.0g of dipotassium hydrogen phosphate, 2.0g of ammonium citrate, 5.0g of anhydrous sodium acetate, 0.5g of magnesium sulfate, 0.25g of manganese sulfate monohydrate, 1.0L of deionized water and pH6.5 (2% agar is added as MRS solid culture medium).
And (3) strain identification:
(1) Morphological observation of bacteria
Gram staining and catalase test are carried out on the screened and purified strain YYS-EN2, physiological and biochemical indexes are measured, and the test result is compared with the eighth edition of Bojie's system bacteriology handbook to carry out preliminary judgment on strains. As shown in FIG. 1, the test shows that the screened strain YYS-EN2 is gram-stained purple and positive. The shape of the material is rod-shaped, the contact enzyme is negative, and no spore is formed.
Extracting YYS-EN2 DNA according to the instruction of the bacterial gene DNA extraction kit, and performing PCR amplification:
PCR amplification process:
Amplifying the 16S rDNA gene sequence by using a primer 27F (5'-AGAGTT TGATCCTGGCTCAG-3') 1492R (5'-GGTTACCTTGTTACGACTT-3'); PCR reaction system: 27F (10 uM) 1 uL, 1492R (10 uM) 1 uL, 10XEasyTag@R Buffer (2.5 mM) 5U L, dNTPs (2.5 mM) 4U L, DNA template 1 uL, easy tag@R DNA Polymerase (5U/L) 0.3U L, ddH 2 0.37.7 uL. PCR amplification procedure: 94℃for 5min, 94℃for 30s, 55℃for 30s, 72℃for 90s, 72℃for 10min, and 32 cycles from the second to the fourth step, 4 ℃.
③ PCR product detection and sequencing analysis, namely, the amplified product is sent to the Guangzhou Qingzhou biological engineering Co Ltd for sequencing, and the strain YYS-EN2 is identified as lactobacillus paracasei.
Wherein, the 16S rDNA gene sequence is as follows:
GCAGTCGAACGAGTTCTCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGG
GTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCC
AACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCC
GGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAA
CCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAG
CGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGAC
GCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGAT
GAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTG
GGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATG
TGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCTTTTGATCACCTGAGAGAT
CAGGTTTCCCCTTCGGGGGCAAAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGAT
GTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGACTAGTTGCCAGCATTTAGTTGGGCACTCT
AGTAAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGA
CCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAGACCGCGAGGTCAAGCTAATC
TCTTAAAGCCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGT
AATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCAT
GAGAGTTTGTAACACCCGAAGCCGGTGGCGTAACCCTTTTAGGGAGCGAGCCGTCTAA。
EXAMPLE 2 preparation of Lactobacillus paracasei YYS-EN2 lyophilized powder
Inoculating activated Lactobacillus paracasei YYS-EN2 into a culture medium sterilized at 121deg.C for 15min according to 2% (v/v), culturing at 37deg.C for 24 hr, centrifuging at 4deg.C for 10min at 6000r/min, discarding supernatant to obtain bacterial mud, emulsifying with protective agent for 15min to obtain emulsion with bacterial solution concentration of 5× 10CFU·mL-1, pre-freezing the emulsion in a refrigerator at-40deg.C for 4 hr, and freeze-drying at-35deg.C for 35 hr to obtain lyophilized powder of active Lactobacillus paracasei YYS-EN2 with living bacterial count of 5100 hundred million CFU.g -1.
Wherein the culture medium comprises the following components in percentage by mass: 2.5% glucose, 2.5% soy peptone, 1% yeast extract, 1% beef extract, 0.6% anhydrous sodium acetate, 0.05% magnesium sulfate, 0.03% manganese sulfate, 0.1% tween 80 and the balance water. The protective agent consists of 50 g.L -1 skimmed milk powder, 10 g.L -1 sucrose, 10 g.L -1 trehalose and 10 g.L -1 L-sodium glutamate.
It should be noted that: the protectant may employ existing protectant components or formulations, including but not limited to the embodiments described above.
EXAMPLE 3 Lactobacillus paracasei YYS-EN2 cellulase Activity
1 Enzyme-producing medium (g.L -1): 20g of sodium carboxymethylcellulose (CMC-Na), 3g of peptone, 0.5g of yeast extract, 2g of ammonium sulfate, 4g of monopotassium phosphate, 0.3g of magnesium sulfate and 0.3g of calcium chloride;
2 measurement of cellulase Activity
(1) Definition of enzyme activity:
the amount of enzyme required to decompose 10 g.L -1 sodium carboxymethylcellulose (CMC-Na) substrate to 1mg glucose per minute at 50℃and pH4.5 was defined as one enzyme activity unit (U.mL -1).
(2) And (3) preparation of a reagent:
0.1 mol.L -1 acetic acid-sodium acetate buffer (pH 4.6): 0.1 mol.L -1 sodium acetate solution and 0.1 mol.L -1 acetic acid solution were mixed and the pH was adjusted to 4.5. Wherein, 0.1 mol.L -1 sodium acetate solution, accurately weighing 8.20g anhydrous sodium acetate, adding distilled water and fixing the volume to a 1000mL volumetric flask; 0.1 mol.L -1 acetic acid solution, accurately weighing 6g acetic acid, adding distilled water and fixing the volume to a 1000mL volumetric flask;
Sodium carboxymethyl cellulose (CMC-Na) solution (10 g·l -1): 1g of sodium carboxymethylcellulose (CMC-Na) was accurately weighed and added to 100mL of acetic acid-sodium acetate buffer.
(3) Enzyme activity determination: the cellulase activity was measured by DNS.
1% Sodium carboxymethylcellulose (CMC-Na) solution is placed in a water bath kettle at 40 ℃ in advance for preheating, 1mL of enzyme solution is added into 1mL of sodium carboxymethylcellulose solution, 1mL of 0.4 mol.L - 1 NaOH is added for stopping the reaction after the reaction is carried out for 30min at 40 ℃, and a stop solution is added before the enzyme solution is added into a blank group.
After the reaction, 1mL of the reaction solution is taken and mixed with 1mL of DNS (5- (dimethylamino) -1-naphthalene sulfonic acid hydrate) solution, and the mixture is reacted in a boiling water bath for 5min for color development. Diluting the reaction solution after color development by 5 times, zeroing by taking distilled water as a reference, measuring the absorbance of a sample at the wavelength of 540nm by using a spectrophotometer, and calculating the enzyme activity according to a formula.
The crude enzyme solution obtaining process in this embodiment is as follows: inoculating Lactobacillus paracasei YYS-EN2 into enzyme production culture medium, culturing at 37deg.C for 48 hr, centrifuging at 4deg.C, and collecting supernatant to obtain crude enzyme solution; the crude enzyme solution was diluted to an enzyme solution at a multiple to perform an enzyme activity assay, wherein Lactobacillus paracasei YYS-EN2 was inoculated in an amount of 2% (v/v) of the medium.
The result measuring and calculating mode is as follows:
The enzyme activity measurement formula is: enzyme activity = OD 540 x n x 1/k x 1/30;
where n is the dilution factor in enzyme solution testing (dilution factor from crude enzyme solution to test enzyme solution), k is the slope of the fitted standard curve, and 30 is the enzymatic reaction time.
Wherein, the standard curve is drawn: standard solutions (0 g.L -1、0.2g·L-1、0.4g·L-1、0.6g·L-1、0.8g·L-1、1.0g·L-1) were prepared in different concentration gradients from 1 g.L -1 glucose solutions, and absorbance at 540nm was measured according to the enzyme activity assay method. A linear standard curve was plotted with glucose concentration on the abscissa and OD on the ordinate (see FIG. 2).
The measurement result shows that the cellulase activity of Lactobacillus paracasei YYS-EN2 is 47.34+ -0.06 U.multidot.mL -1.
EXAMPLE 4 Lactobacillus paracasei YYS-EN2 pectase Activity
1. Enzyme-producing medium (g.L -1): 10g of pectin, 10g of glucose, 4g of peptone, 4g of dipotassium hydrogen phosphate, 2g of potassium dihydrogen phosphate and 0.5g of magnesium sulfate heptahydrate;
2. Pectase activity assay
(1) Definition of enzyme activity:
the amount of enzyme required to decompose 10 g.L -1 of pectin substrate to 1mg of galacturonic acid per minute at 50℃and pH 5.0 is defined as one enzyme activity unit (U.mL -1);
(2) Preparing a solution:
0.2 mol.L -1 acetic acid-sodium acetate buffer (pH 4.8): 0.2 mol.L -1 sodium acetate solution and 0.2 mol.L -1 acetic acid solution were mixed and the pH was adjusted to 5.0. Wherein, 0.2 mol.L -1 of acetic acid solution: accurately weighing 12g of acetic acid, adding distilled water and fixing the volume to a 1000mL volumetric flask; 0.2 mol.L -1 of sodium acetate solution, accurately weighing 16.41g of sodium acetate, adding distilled water and fixing the volume to a 1000mL volumetric flask. The solution was mixed and the pH was adjusted to 5.0.
Pectin substrate (10 g.L -1): 1g pectin is added to 100mL acetic acid-sodium acetate buffer
(3) Enzyme activity determination:
Placing pectin substrate in a water bath kettle at 50 ℃ in advance for preheating; 1mL of enzyme solution is taken, 1mL of preheated pectin substrate is added, the mixture is shaken uniformly, the mixture is reacted in a water bath at 50 ℃ for 30min, 1mL of 10% trichloroacetic acid solution is added for stopping the reaction, and a stop solution is added before the enzyme solution is added in a blank group. After the reaction is finished, 1mL of reaction solution is taken and mixed with 1mLDNS solution uniformly, and the mixture is reacted in boiling water bath for 5min for color development. Diluting the reaction solution after color development by 5 times, zeroing by taking distilled water as a reference, measuring the absorbance of a sample at the wavelength of 540nm by using a spectrophotometer, and calculating the enzyme activity according to a formula.
The crude enzyme solution obtaining process in this embodiment is as follows: inoculating Lactobacillus paracasei YYS-EN2 into enzyme production culture medium, culturing at 37deg.C for 48 hr, centrifuging at 4deg.C, and collecting supernatant to obtain crude enzyme solution; the crude enzyme solution was diluted to an enzyme solution at a multiple to perform an enzyme activity assay, wherein Lactobacillus paracasei YYS-EN2 was inoculated in an amount of 2% (v/v) of the medium.
The result measuring and calculating mode is as follows:
The enzyme activity measurement formula is: enzyme activity = OD 540 x n x 1/k x 1/30;
where n is the dilution factor in enzyme solution testing (dilution factor from crude enzyme solution to test enzyme solution), k is the slope of the fitted standard curve, and 30 is the enzymatic reaction time.
Wherein, the standard curve is drawn: standard solutions (0 g.L -1、0.2g·L-1、0.4g·L-1、0.6g·L-1、0.8g·L-1、1.0g·L-1) were prepared in different concentration gradients from 1 g.L -1 of D-galacturonic acid solution, and absorbance at 540nm was measured according to the enzyme activity assay method. A linear standard curve was plotted with D-galacturonic acid concentration on the abscissa and OD on the ordinate (see FIG. 3).
The measurement result shows that the pectase activity of Lactobacillus paracasei YYS-EN2 is 47.81 + -0.07U/mL.
Example 5 Lactobacillus paracasei YYS-EN2 tannin enzyme Activity
1. Enzyme-producing medium (g.L -1): 10g of glucose, 3g of sodium nitrate, 1g of dipotassium hydrogen phosphate, 0.5g of potassium chloride, 0.5g of magnesium sulfate and 20g of tannic acid.
2. Tannin enzyme activity assay
(1) Definition of enzyme activity:
The amount of enzyme required to decompose the substrate 0.01 mol.L -1 propyl gallate per minute to produce 1mg of gallic acid at 30℃and pH 5.0 was defined as one enzyme activity unit (U.mL -1);
(2) Preparing a solution:
0.1 mol.L -1 citric acid-sodium citrate buffer (pH 5.0): 0.1 mol.L -1 of citric acid solution and 0.1 mol.L -1 of sodium citrate solution were mixed and the pH was adjusted to 5.0. Wherein, 0.1 mol.L -1 citric acid solution: accurately weighing 19.21g of citric acid, and fixing the volume to a 1000mL volumetric flask by using distilled water; sodium citrate solution of 0.1mol·l -1: 29.41g sodium citrate was accurately weighed, dissolved in distilled water and fixed to a 1000mL volumetric flask.
Propyl gallate solution (0.01 mol·l -1): accurately weighing 0.21g propyl gallate, and fixing the volume to a 100mL volumetric flask by using a citric acid buffer solution.
Methanol rhodamine solution: accurately weighing 0.667g of rhodanine, and fixing the volume to a 100mL volumetric flask by using methanol.
Gallic acid standard solution (5 mg. L -1): accurately weighing 0.5g gallic acid, and fixing the volume to a 100mL volumetric flask by using methanol.
(3) Enzyme activity determination:
Preheating propyl gallate solution in a water bath kettle at 30 ℃ in advance; 0.5mL of the enzyme solution is taken, 0.5mL of preheated propyl gallate solution is added, the mixture is shaken uniformly, the mixture is reacted in a water bath at 30 ℃ for 5min, 0.6mL of methanol rhodanine is added, the water bath at 30 ℃ for 5min, 0.8mL of KOH is added, the water bath at 30 ℃ for 5min, 7.6mL of distilled water is added, and the water bath at 30 ℃ for 10min. Zeroing by taking distilled water as a reference, measuring the absorbance of a sample at a wavelength of 520nm by using a spectrophotometer, and calculating the enzyme activity according to a formula. The blank group is added with a stop solution KOH before the enzyme solution is added.
The crude enzyme solution obtaining process in this embodiment is as follows: inoculating Lactobacillus paracasei YYS-EN2 into an enzyme production culture medium, culturing at 37deg.C for 48 hr, centrifuging at 4deg.C, collecting supernatant to obtain crude enzyme solution, and diluting the crude enzyme solution to enzyme solution for testing according to a certain multiple to perform enzyme activity measurement; wherein, the inoculation amount of lactobacillus paracasei YYS-EN2 is 2% (v/v) of the culture medium.
The result measuring and calculating mode is as follows:
The enzyme activity measurement formula is: enzyme activity = OD 520 x n x 1/k x 1/5 x 1/0.5;
Wherein n is the dilution factor (dilution factor of crude enzyme solution to that of enzyme solution for test or that of reaction solution after color development) in enzyme solution test, k is the slope of a fitted standard curve, 5 is the enzymatic reaction time, and 1/0.5 is converted into 1mL of enzyme solution.
Wherein, the standard curve is drawn: standard solutions (0mg.L -1、1mg·L-1、2mg·L-1、3mg·L-1、4mg·L-1、5mg·L-1) with different concentration gradients were prepared from 5 mg.L -1 gallic acid solution, and absorbance at 520nm was measured according to the enzyme activity assay method. Drawing a linear standard curve (see FIG. 4) with gallic acid concentration on the abscissa and OD value on the ordinate
The measurement result showed that the tannase activity of Lactobacillus paracasei YYS-EN2 was 29.85.+ -. 0.03 U.mL -1.
EXAMPLE 6 Lactobacillus paracasei YYS-EN2 Phytase Activity
1. Enzyme-producing medium (g.L -1): 3g of peptone, 15g of glucose, 0.5g of magnesium sulfate, 0.03g of manganese sulfate and 0.03g of ferrous sulfate.
2. Phytase Activity assay
(1) Definition of enzyme activity:
1mL of liquid enzyme, at 37 degrees C, pH 5.5 conditions per minute to decompose 50 mmol L -1 sodium phytate substrate, to produce 1U mo L inorganic phosphorus, namely a phytase activity unit, to represent U.mL -1.
(2) And (3) preparation of a reagent:
0.2 mol.L -1 acetic acid-sodium acetate buffer (pH 5.5): 0.2 mol.L -1 acetic acid solution and 0.2 mol.L -1 sodium acetate solution were mixed and the pH was adjusted to 5.5. Wherein, 0.2 mol.L -1 of acetic acid solution: 12g acetic acid is fixed to volume by distilled water into a 1000mL volumetric flask; 0.2 mol.L -1 sodium acetate solution; 16.406g of sodium acetate are dissolved in distilled water and are taken up in 1000mL volumetric flasks.
Sodium phytate solution (75 mmol. L -1): accurately weighing 0.69g of sodium phytate, dissolving the sodium phytate by using a buffer solution, and fixing the volume into a 100mL volumetric flask.
Nitric acid solution: nitric acid: water (v/v) =1: 2
Ammonium molybdate solution (100 g/L) 10g ammonium molybdate was accurately weighed, added to 1mL 25% (v/v) ammonia water and distilled water was used to volume to a 100mL volumetric flask.
The ammonium metavanadate solution (2.35 g/L) is prepared by accurately weighing 0.235g of ammonium metavanadate, dissolving in distilled water, adding 2mL of nitric acid solution, and using distilled water to fix the volume to a 100mL volumetric flask, and storing in a dark place for use.
Color development liquid (stop liquid): nitric acid solution: ammonium molybdate solution: ammonium metavanadate solution (v/v) =2: 1:1, storing in a dark place, and preparing at present.
(3) Enzyme activity determination:
firstly, 1.8mL of acetic acid buffer solution is mixed with 0.2mL of to-be-reacted liquid (namely enzyme liquid), the mixture is preheated for 5min at 37 ℃ in a water bath kettle, then 4mL of sodium phytate is added for uniform mixing, water bath is carried out for 30min at 37 ℃, and finally 4mL of chromogenic liquid (stop solution) is added for uniform mixing. Zeroing by taking distilled water as a reference, measuring the absorbance of a sample at 415nm by using a spectrophotometer, and calculating the enzyme activity according to a formula. The blank group is added with stop solution before enzyme solution is added.
The crude enzyme solution obtaining process in this embodiment is as follows: inoculating Lactobacillus paracasei YYS-EN2 into an enzyme production culture medium, culturing at 37deg.C for 48 hr, centrifuging at 4deg.C, collecting supernatant to obtain crude enzyme solution, and diluting the crude enzyme solution to enzyme solution according to a certain multiple for enzyme activity determination; wherein, the inoculation amount of lactobacillus paracasei YYS-EN2 is 2% (v/v) of the culture medium.
The result measuring and calculating mode is as follows:
The enzyme activity measurement formula is: enzyme activity = OD 415×n×1/k×1/30×1/0.2×103;
Wherein n is the dilution factor (from crude enzyme to enzyme for test) in enzyme solution test, k is the slope of a fitted standard curve, 30 is the enzymatic reaction time, 1/0.2 is converted into 1mL of enzyme solution, and 10 3 is the conversion factor of inorganic phosphorus ion concentration.
Wherein, the standard curve is drawn: inorganic phosphorus standard solutions (0mmo l·L-1、10mmo l·L-1、20mmo l·L-1、30mmo l·L-1、40mmo l·L-1、50mmo l·L-1), with different concentrations are prepared from potassium dihydrogen phosphate mother solution, the light absorption value of OD 415 is measured according to an enzyme activity measuring method, the inorganic phosphorus ion concentration is taken as an abscissa, the OD value is taken as an ordinate, and a linear standard curve is drawn (see FIG. 5).
The measurement result showed that the phytase activity of Lactobacillus paracasei YYS-EN2 was 18.67.+ -. 0.04U/mL.
Example 7 use of Lactobacillus paracasei YYS-EN2 in the preparation of fermented corn nutritional powder
Crushing corn embryo and corn husk, treating with lipase and amylase, sterilizing at high temperature, adding lactobacillus paracasei YYS-EN2 bacterial powder and/or freeze-dried powder 3% (by mass), mixing, fermenting at 37deg.C for 72 hr, sterilizing at high temperature after fermentation, and drying at 60deg.C to obtain corn nutrition powder. Lactobacillus paracasei YYS-EN2 ferments corn husks and corn embryos, breaks down cellulose and pectin and reduces tannins and phytic acid, enhancing the flavor and mouthfeel of the corn nutritional powder. The flavor results before and after the specific fermentation are shown in Table 1 below:
TABLE 1
Example 12 use of Lactobacillus paracasei YYS-EN2 in preparation of fermented traditional Chinese medicine
Pulverizing rhizoma Polygonati, ginseng radix (artificially cultivated), fructus Lycii, mori fructus and fructus Jujubae, and adding at a ratio of 0.5%, 0.3%, 1%, 1.5% and 2% by mass respectively; mixing uniformly, boiling, and keeping micro boiling for 1h; cooling, adding lactobacillus paracasei YYS-EN2 powder and/or lyophilized powder 0.03% and lactobacillus plantarum BXM2 powder 0.02%, mixing, fermenting at 37deg.C for 48 hr, and sterilizing at high temperature. Lactobacillus paracasei YYS-EN2 ferments the Chinese medicine, decomposes cellulose and pectin, reduces tannin and phytic acid, reduces bitter taste and enhances the mouthfeel.
The Chinese medicine is not limited to the specific Chinese medicine in the embodiment, and comprises the types in the medicine-food homology list. The probiotic combination comprises but is not limited to lactobacillus paracasei YYS-EN2 bacterial powder and lactobacillus plantarum BXM2 bacterial powder, and can also be independently used by lactobacillus paracasei YYS-EN2 bacterial powder or used together with other probiotic powder.
Wherein, the lactobacillus plantarum (Lactobacillus plantarum) BMX2 is adopted and is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) in 9 and 6 days of 2018, the preservation address is North Star Xiyu No.1 and 3 in the Korean area of Beijing city, and the preservation number is CGMCC No.16436.
According to the results of the above examples, lactobacillus paracasei YYS-EN2 provided by the present invention has the following properties and effects:
1. Has the capability of producing cellulase, and the cellulase activity is 47.34+/-0.06 U.mL -1; can decompose plant cellulose and can be widely applied to the development of fermented traditional Chinese medicine and silage products;
2. Has pectase capability, and pectase activity is 47.81 +/-0.07 U.mL -1; can decompose pectin in traditional Chinese medicines, fruits, vegetables and the like, and can be widely applied to the development of products such as fermented traditional Chinese medicines, fruit juice, vegetables, wines, enzymes and the like;
3. Has tannase capability, and the tannase activity is 29.85+/-0.03 U.mL -1; the tannin content in the fermented traditional Chinese medicine and wine can be reduced, and the method can be widely applied to the development of fermented traditional Chinese medicine, wine and ferment products, and can improve the taste of the products;
4. Has phytase ability, and the phytase activity is 18.67+/-0.04 U.mL -1; can effectively decompose phytic acid, can be widely applied to flour and soybean food processing, and improves the nutrition and commodity value of the flour and soybean food;
5. can be used for preparing corn nutrition powder, degrading cellulose and phytic acid in corn husks and corn embryos, improving the nutrition value of the corn nutrition powder and improving the taste;
6. the strain is derived from oral saliva of healthy adults, is natural probiotics, and can be used as a starter for preparing fermented traditional Chinese medicines, feeds, wine, ferment and the like.
In summary, compared with the prior art, the lactobacillus paracasei YYS-EN2 provided by the invention has the following beneficial effects:
The lactobacillus paracasei YYS-EN2 provided by the invention can be used for probiotic products and functional foods, and can be planted in human intestinal tracts to play roles in improving flora structures and promoting digestion and absorption. The method can produce cellulase, pectase, tannase and phytase, and can be applied to probiotic fermentation products (fermentation products comprise but are not limited to fermentation traditional Chinese medicines, beverages, foods, feeds and the like), wherein the fermentation products comprise but are not limited to preparation of fermentation traditional Chinese medicines, fermentation plant-derived nutrition powder, fermentation plant-derived polypeptide powder, fermentation plant-derived protein powder, ferment, silage and the like;
The lactobacillus paracasei YYS-EN2 provided by the invention can produce cellulase and pectase, can degrade plant cellulose and pectin, and can be applied to development of products such as fermented traditional Chinese medicines, fermented plant-derived nutrition powder, fruit juice, vegetable feed and the like; meanwhile, the strain has high pectase and tannase capabilities, can be widely applied to the production of fermented products such as fermented traditional Chinese medicines, wines, enzymes and the like, and can obviously reduce the content of tannin in the products.
In summary, it is particularly applicable in functional products comprising at least one of the following actions:
(1) Has cellulose degradation capability;
(2) Has pectin degradation capability;
(3) Has tannin degradation capability;
(4) Has the capability of degrading phytic acid;
The products with the functions of (1) - (4) comprise, but are not limited to, the functions of decomposing plant fibers and pectin, improving the clarity of the products and the like, and can also be products with other obvious effects based on the functions of decomposing plant fibers and pectin and degrading tannins or phytic acid.
It should be noted that:
(1) Definition:
the term "food" as used herein is broad and includes human foods and drinks. In certain embodiments, the food product is suitable and designed for human feeding. The application can be used for preparing solid preparations such as powder, tablets, gel and the like, and also can be dispersed in liquid to prepare liquid preparations, including but not limited to the embodiment.
The traditional Chinese medicine in the "fermented traditional Chinese medicine product" described herein comprises the species contained in the catalogue of homology of medicine and food.
(2) Related prior art means or prior art terms to which the present application relates:
The "OD" value is an abbreviation of optical density (also called absorbance), and the difference between the energy of light passing through the object to be measured and the energy of light passing through the object to be measured is the energy absorbed by the object to be measured. "OD X" is an optical density value measured when the wavelength is set to Xnm, and is a standard indicator for tracking the density of microorganisms in a liquid culture, and is generally used to indicate the cell density of a cell. Among these, the "OD" value determination method is prior art, and the principle and method thereof will not be described here.
The oxford cup method is used in the prior art, and the process is not tired. (3) Strain application:
The examples illustrate the application of lactobacillus paracasei YYS-EN2 to corn meal products, which can be applied to various types of products containing cellulose, pectin, tannin and phytic acid according to the design concept described above, and can be applied to the preparation of fermented products including fermented traditional Chinese medicines, fermented plant-derived nutrition powder, fermented plant-derived polypeptide powder, fermented plant-derived protein powder, ferment, silage and the like, including but not limited to corn meal in the examples.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (10)
1. Lactobacillus paracasei YYS-EN2 is characterized in that the preservation number is CGMCC No.28531.
2. Lactobacillus paracasei YYS-EN2 according to claim 1, characterized in that: it has the ability to produce cellulases, pectinases, tannase and phytases.
3. Lactobacillus paracasei YYS-EN2 according to claim 1, characterized in that: it has the ability to degrade cellulose, pectin, tannins and phytic acid.
4. A method for producing cellulases, pectinases, tannase and phytases, characterized by: inoculating lactobacillus paracasei YYS-EN2 into an enzyme-producing liquid culture medium, culturing at the temperature of between 30 and 40 ℃ for 48 to 72 hours, and carrying out solid-liquid separation on the enzyme-producing liquid culture medium after fermentation culture to obtain a supernatant; the supernatant is an enzyme solution containing cellulase and/or pectase and/or tannase and/or phytase;
Wherein the Lactobacillus paracasei YYS-EN2 is Lactobacillus paracasei YYS-EN2 according to any of claims 1-3.
5. The application of lactobacillus paracasei YYS-EN2 in preparing a fermentation product is characterized in that: the lactobacillus paracasei YYS-EN2 employs the lactobacillus paracasei YYS-EN2 as claimed in any of claims 1-3.
6. Use of lactobacillus paracasei YYS-EN2 according to claim 5 for the preparation of a fermented product, characterized in that: the fermentation product comprises fermented traditional Chinese medicines, fermented nutrition powder, fermented polypeptide powder, fermented protein powder, ferment and silage;
The fermentation nutrition powder comprises fermentation plant source nutrition powder; the fermented polypeptide powder comprises fermented plant-derived polypeptide powder; the fermented protein powder comprises fermented plant-derived protein powder.
7. A lyophilized product, characterized in that: a component comprising lactobacillus paracasei YYS-EN2 as claimed in any of claims 1 to 3.
8. A microbial inoculum, characterized in that: the components of the microbial inoculum comprise the freeze-dried powder of claim 7; and/or the components of the microbial inoculum comprise lactobacillus paracasei YYS-EN2 as claimed in any one of claims 1-3.
9. The application of lactobacillus paracasei YYS-EN2 in the preparation of a probiotic product is characterized in that: the lactobacillus paracasei YYS-EN2 employs the lactobacillus paracasei YYS-EN2 as claimed in any of claims 1-3.
10. The application of lactobacillus paracasei YYS-EN2 in the preparation of functional products is characterized in that: the functional product comprises at least one of the following functions:
(1) Has cellulose degradation capability;
(2) Has pectin degradation capability;
(3) Has tannin degradation capability;
(4) Has the capability of degrading phytic acid;
Wherein the Lactobacillus paracasei YYS-EN2 is Lactobacillus paracasei YYS-EN2 according to any of claims 1-3.
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