CN105925499A - Pediococcus acidilactici JQQ2 and applications thereof - Google Patents
Pediococcus acidilactici JQQ2 and applications thereof Download PDFInfo
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- CN105925499A CN105925499A CN201610281244.0A CN201610281244A CN105925499A CN 105925499 A CN105925499 A CN 105925499A CN 201610281244 A CN201610281244 A CN 201610281244A CN 105925499 A CN105925499 A CN 105925499A
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- 241000191998 Pediococcus acidilactici Species 0.000 title claims abstract description 24
- 238000004321 preservation Methods 0.000 claims abstract description 7
- 235000021001 fermented dairy product Nutrition 0.000 claims abstract description 5
- 238000002360 preparation method Methods 0.000 claims description 6
- 230000033228 biological regulation Effects 0.000 claims description 5
- 210000000936 intestine Anatomy 0.000 claims description 2
- 210000002784 stomach Anatomy 0.000 claims description 2
- 239000000654 additive Substances 0.000 claims 1
- 230000000996 additive effect Effects 0.000 claims 1
- 235000013305 food Nutrition 0.000 claims 1
- 238000009629 microbiological culture Methods 0.000 claims 1
- 235000013373 food additive Nutrition 0.000 abstract description 5
- 239000002778 food additive Substances 0.000 abstract description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 5
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract 2
- 238000012360 testing method Methods 0.000 description 21
- 241000894006 Bacteria Species 0.000 description 20
- 230000001580 bacterial effect Effects 0.000 description 16
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 8
- 239000002609 medium Substances 0.000 description 7
- 230000003385 bacteriostatic effect Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 238000012408 PCR amplification Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 239000004310 lactic acid Substances 0.000 description 4
- 235000014655 lactic acid Nutrition 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000000386 microscopy Methods 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 108010062877 Bacteriocins Proteins 0.000 description 2
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L calcium carbonate Substances [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000012797 qualification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- DBOVHQOUSDWAPQ-UHFFFAOYSA-N (4aS)-6c-[O2-(3,5,3'-trihydroxy-biphenyl-2-carbonyl)-beta-D-glucopyranosyloxy]-5t-vinyl-(4ar)-4,4a,5,6-tetrahydro-3H-pyrano[3,4-c]pyran-1-one Natural products OC1C(O)C(CO)OC(OC2C(C3C(C(OCC3)=O)=CO2)C=C)C1OC(=O)C1=C(O)C=C(O)C=C1C1=CC=CC(O)=C1 DBOVHQOUSDWAPQ-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- BZXINCMCFVKGKB-UHFFFAOYSA-N Amarogentin Natural products OCC1OC(OC2OC=C3C(CCOC3=O)C2C=C)C(OC(=O)c4cc(O)cc(O)c4c5cccc(O)c5)C(O)C1O BZXINCMCFVKGKB-UHFFFAOYSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241000191938 Micrococcus luteus Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 241000896292 Odontothrips loti Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- MUPFEKGTMRGPLJ-RMMQSMQOSA-N Raffinose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 MUPFEKGTMRGPLJ-RMMQSMQOSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- MUPFEKGTMRGPLJ-UHFFFAOYSA-N UNPD196149 Natural products OC1C(O)C(CO)OC1(CO)OC1C(O)C(O)C(O)C(COC2C(C(O)C(O)C(CO)O2)O)O1 MUPFEKGTMRGPLJ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000020167 acidified milk Nutrition 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- DBOVHQOUSDWAPQ-WTONXPSSSA-N amarogentin Chemical compound O([C@H]1[C@H](O[C@H]2[C@@H]([C@H]3C(C(OCC3)=O)=CO2)C=C)O[C@@H]([C@H]([C@@H]1O)O)CO)C(=O)C1=C(O)C=C(O)C=C1C1=CC=CC(O)=C1 DBOVHQOUSDWAPQ-WTONXPSSSA-N 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- MUPFEKGTMRGPLJ-ZQSKZDJDSA-N raffinose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O2)O)O1 MUPFEKGTMRGPLJ-ZQSKZDJDSA-N 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
-
- 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
- A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/41—Pediococcus
- A23V2400/413—Acidilactici
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (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)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides pediococcus acidilactici JQQ2, wherein the classification name of pediococcus acidilactici JQQ2 is pediococcus acidilactici, the preservation number is CGMCC No.11929, and the preservation date is Dec 24, 2015. The invention further provides applications of the strain in preparing food additives of fermented dairy products and products for regulating the microecological balance of the gastrointestinal tracts. The obtained strain has excellent antibacterial property, as well as excellent high-temperature resistance and excellent extreme pH resistance. The strain has good application prospects in preparing the food additives of the fermented dairy products and the products for regulating the microecological balance of the gastrointestinal tracts.
Description
Technical field
The invention belongs to microorganism field, relate to strain Pediococcus acidilactici JQQ2 and an application thereof,
It is specifically related to a strain and high temperature and wide in range pH are had the Pediococcus acidilactici JQQ2 of outstanding tolerance
And application.
Background technology
In recent years, owing to functional food is increasingly subject to pay attention to, people's research and development enthusiasm to probio
The most surging.Wherein, Pediococcus acidilactici is important one.But, the breast of current gained
The resistance ability of acid sheet coccus is the most unsatisfactory, is mainly manifested in high temperature and extreme pH
Tolerance is poor.Such as " Pediococcus acidilactici PA003 bacteriostasis and the research of external tolerance "
Reporting a strain Pediococcus acidilactici PA003, it is the most poor to the tolerance of high temperature.For another example
《Characterization of two bacteriocins produced by Pediococcus
acidilactici isolated from“Alheira”,a fermented sausage traditionally
Produced in Portugal " the two strain Pediococcus acidilacticis announced are equal to the tolerance of pH and temperature
Poor.
Therefore, seek a strain all have good anti-microbial property and have outstanding resistance (especially for
High temperature and extreme pH) Pediococcus acidilactici, become this area problem demanding prompt solution.
Summary of the invention
For the shortcoming of prior art, an object of the present invention is to provide a strain lactic acid sheet ball
Bacterium JQQ2, the Classification And Nomenclature of this Pediococcus acidilactici JQQ2 is Pediococcus acidilactici Pediococcus
Acidilactici, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center
CGMCC (Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences's microbe research
Institute's postcode 100101), preserving number is CGMCC No.11929, and the preservation time is 2015
On December 24, in.
As shown in the embodiment of the present invention, the resistance to elevated temperatures of obtained strains of the present invention is excellent, when
After processing 30min in 121 DEG C, the bacteriostatic activity retention rate of bacterial strain is still up to 81.09%.With
Time, obtained strains of the present invention is also fine for the tolerance of extreme pH, when pH=2,
Bacteriostatic activity retention rate is up to 81.3%, and under pH=10, bacteriostatic activity only loses 39.86%.
Therefore, one of contribution for prior art of the present invention there are provided a strain for height
Gentle extreme pH has the Pediococcus acidilactici of outstanding tolerance, has expanded answering of Pediococcus acidilactici
By scope and potentiality.
Based on the above-mentioned outstanding characteristic of bacterial strain of the present invention, those skilled in the art easily combine ability
The routine techniques means in territory are applied in terms of preparing fermented dairy product food additives.Cause
This, further object is that offer Pediococcus acidilactici JQQ2 is preparing acidified milk system
Savor the application in terms of food additives.
Based on the above-mentioned outstanding characteristic of bacterial strain of the present invention, those skilled in the art easily combine ability
The routine techniques means in territory are applied in preparation preparation regulation micro ecology of gastrointestinal tract equilibrium product
Aspect.Therefore, further object is that offer Pediococcus acidilactici JQQ2 is in preparation
Application in terms of fermentation preparation regulation micro ecology of gastrointestinal tract equilibrium product.
Beneficial effects of the present invention:
1, obtained strains of the present invention has outstanding anti-microbial property；
2, obtained strains of the present invention has outstanding resistance to elevated temperatures, when processing 30 in 121 DEG C
After min, the bacteriostatic activity retention rate of bacterial strain is still up to 81.09%；
3, obtained strains of the present invention has outstanding resistance to extreme pH performance, when pH=2, presses down
Bacterium activity preservation rate is up to 81.3%, and under pH=10, bacteriostatic activity only loses 39.86%；
4, obtained strains of the present invention is preparing fermented dairy product food additives and regulation intestines and stomach
Microecological balance product aspect has a good application prospect.
Accompanying drawing explanation
Fig. 1 is the microscopy form result figure of JQQ2；
Fig. 2 is the PCR result figure of JQQ2.
Detailed description of the invention
Below by embodiment, the present invention is specifically described, it is necessary to it is pointed out here that be with
Lower embodiment is only intended to be further detailed the present invention, it is impossible to be interpreted as the present invention
The restriction of protection domain, the person skilled in the art in this field is made according to foregoing invention content
Some nonessential improvement and adjustment, still fall within protection scope of the present invention.
Embodiment 1
1, experimental raw and method
1.1 bacterium source
Homemade traditional natural fermentation pickled vegetable.
1.2 culture medium
MRS culture medium: beef extract (powder) 10g, peptone 10g, yeast extract (powder) 5g,
Glucose 20g, Tween-80 1mL, ammonium citrate 2g, K2HPO42g, sodium acetate 5g,
(preparation solid medium need to add for magnesium sulfate 0.58g, manganese sulfate 0.25g, distilled water 1000mL
Agar powder 18g), heating for dissolving, regulate pH to 6.0～6.4,121 DEG C of sterilizing 15min.
1.3 16S rDNA sequence analyses
The explanation extracting kit according to bacterial genomes extracts bacterial strain DNA, with DNA as mould
Plate, PCR expands its 16S rDNA fragment.Upstream primer P1:
5 '-AGAGTTTGATCCTGGCTCAG-3 ', downstream primer P2:
5’-CTACGGCTACCTTGTTACGA-3’。
PCR amplification system and amplification program be as shown in Table 1 and Table 2:
Table 1 PCR amplification system
Table 2 PCR amplification program
Take pcr amplification product and DNA Marker DL2000, use 1% Ago-Gel (to contain
Golden View) electrophoresis detection, voltage 100V, horizontal strip electrophoresis 30min, use gel imaging
Instrument is observed.PCR primer entrusts the order-checking of English Weihe River victory base (Shanghai) trade Co., Ltd.16S rDNA
Sequence is compared in NCBI website.
Product after carrying out PCR with JQQ2 genomic DNA for template, through Ago-Gel
Electrophoresis, it is seen that clear band occurs in 1000bp-2000bp region, such as Fig. 2.
The pcr amplification product sequencing result of bacterial strain JQQ2 is passed through BLAST on NCBI website
Instrument is analyzed, result such as table 3:
Table 3
1.4 lactic acid bacterias isolated and purified
Use 10 times of gradient dilution methods that pickles water sample is diluted to 10-5、10-6、10-7Dilute
Degree of releasing.Each dilution factor draws 1mL bacterium solution, and with containing 1.5%CaCO3MRS solid training
Support base pour plate, cultivate 48h, select the bacterium colony of the different shape of obvious molten calcium circle for 37 DEG C
Carrying out Gram's staining observation, the gram-positive bacterium colony of picking contains to another
1.5%CaCO3MRS solid plate on line cultivate, and will obtain on solid medium flat board
Single colony inoculation in a test tube equipped with 5mL MRS fluid nutrient medium, in 37 DEG C
Lower cultivation 24h, then continues line by oese picking bacterium solution, repeats 2-3 time, until bacterium
It is consistent that the microscopy result that falls shows as thalli morphology, can preserve bacterial strain.
The preservation of 1.5 bacterial strains
Activating 2-3 time in the single bacterial strain liquid medium within that will obtain, streak inoculation is in MRS
Solid slope is cultivated 48h and is placed on short-term preservation in 4 DEG C of refrigerators, or by liquid culture with
The sterile glycerol of 40% is mixed in the ratio of 1:1, preserves for a long time under the conditions of-20 DEG C.
The qualification of 1.6 lactic acid bacterias
(1) morphological feature of lactic acid bacteria
By bacterium solution streak inoculation on solid medium, cultivate 48h for 37 DEG C and observe and record bacterium colony
Form, carries out gram stain microscopy simultaneously, observes and record its morphological features.
(2) physiological and biochemical property
Carry out producing H to the bacterial strain of the tolerance digestive tract environment that initial gross separation screening obtains2S test,
Nitrate reduction test, gelatin liquefaction test, indoles produce test, lactose test, mannose
Test, raffinose test, trehalose test, aesculin test, maltose test, fiber two
Sugar test, fructose test, rhamnose test, xylose test, galactolipin test, arabinose
Test, amarogentin test, biochemical anti-with reference to uncle's outstanding Bacteria Identification handbook (the 8th edition) contrast
Should result, and bacterial strain is carried out preliminary population classification.
The colony characteristics of JQQ2 is as shown in table 4 with individual morphology；Microscopy form such as Fig. 1 of JQQ2
Shown in.
Table 4
The Physiology and biochemistry qualification result of bacterial strain JQQ2 is as shown in table 5.
Table 5
Note: "+" be positive；"-" is negative.
1.7 bacteriostatic experiment
Use double-deck agar Odontothrips loti, the plain agar medium of 10mL sterilizing is cooled to
It is poured into sterilized petri dishes after 50～60 DEG C, topples over 10mL after solidification containing 200uL 10-7CFU/mL
The nutrient agar of indicator bacteria suspension.Standing puts into Oxford cup after drying (diameter is about
6mm), each flat board uniformly places 4, and draws 200uL tested bacterium supernatant with liquid-transfering gun
Liquid injects in the cup of Oxford, mark.Shift after culture dish is put into 4 DEG C of refrigerator prediffusion 6～12h
To constant incubator, quiescent culture 24h at 37 DEG C.The bacterial strain choosing obvious inhibition zone does
Next step experiment.
1.8pH tolerance test
Under the conditions of 37 DEG C, the bacterium solution cultivating 12h in test tube is inoculated in by the inoculum concentration of 1%
In 100mLMRS fluid nutrient medium, 37 DEG C of DEG C of quiescent culture 24h.Bacterium solution is pressed the inoculation of 1%
(adjust bacterial concentration is 10 to amount7Cfu/mL) pH=2.0, pH=4.0, pH=7 it are seeded to respectively,
In the MRS culture medium of pH=10, (hydrochloric acid regulation), 37 DEG C of quiescent culture 4h, measure antibacterial work
Property retention rate, it is judged that bacterial strain tolerance situation under sour environment.Often group sets 3 repetitions.
Table 6 is the pH tolerance test result of JQQ2.
Table 6
1.9 heatproof experiments
Bacteriocin after purification is redissolved in aseptic ultra-pure water with 40:1, be respectively placed in 20 DEG C,
40 DEG C, 60 DEG C, 80 DEG C, process 20min under 100 DEG C of water-baths and 121 DEG C (high pressure), with
The sample liquid that 20 DEG C processed is as comparison, with micrococcus luteus as indicator bacteria, makes of Oxford cup and presses down
Bacterium is tested.
Table 7 is the heatproof experimental result of JQQ2.
Table 7
Claims (3)
1. a strain Pediococcus acidilactici JQQ2, it is characterised in that described Pediococcus acidilactici JQQ2
Classification And Nomenclature be Pediococcus acidilactici Pediococcus acidilactici, be preserved in China Microbiological
Culture presevation administration committee common micro-organisms center CGMCC, preserving number is CGMCC
No.11929, the preservation time is on December 24th, 2015.
2. the Pediococcus acidilactici JQQ2 described in claim 1 is preparing fermented dairy product food
Application in terms of additive.
3. the Pediococcus acidilactici JQQ2 described in claim 1 is in the preparation regulation micro-life of intestines and stomach
Application in terms of state equilibrium product.
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| CN117165481B (en) * | 2023-09-05 | 2024-06-04 | 天津科技大学 | Lactobacillus plantarum capable of degrading malic acid and application thereof |
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| CN110343635A (en) * | 2019-06-06 | 2019-10-18 | 四川省食品发酵工业研究设计院 | The Pediococcus acidilactici of one plant of fermentation sauce flavouring |
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| CN117165481B (en) * | 2023-09-05 | 2024-06-04 | 天津科技大学 | Lactobacillus plantarum capable of degrading malic acid and application thereof |
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