CN115011498A - High-temperature-resistant and high-sugar-resistant pediococcus acidilactici - Google Patents
High-temperature-resistant and high-sugar-resistant pediococcus acidilactici Download PDFInfo
- Publication number
- CN115011498A CN115011498A CN202110245170.6A CN202110245170A CN115011498A CN 115011498 A CN115011498 A CN 115011498A CN 202110245170 A CN202110245170 A CN 202110245170A CN 115011498 A CN115011498 A CN 115011498A
- Authority
- CN
- China
- Prior art keywords
- pediococcus acidilactici
- strain
- sugar
- culture medium
- resistant
- 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.)
- Pending
Links
- 241000191998 Pediococcus acidilactici Species 0.000 title claims abstract description 67
- 241000894006 Bacteria Species 0.000 claims description 31
- 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 claims description 29
- 239000008103 glucose Substances 0.000 claims description 29
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose 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)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 19
- 229930006000 Sucrose Natural products 0.000 claims description 19
- 239000005720 sucrose Substances 0.000 claims description 19
- 238000000034 method Methods 0.000 claims description 17
- 238000000855 fermentation Methods 0.000 claims description 15
- 230000004151 fermentation Effects 0.000 claims description 15
- 235000019764 Soybean Meal Nutrition 0.000 claims description 12
- 239000004455 soybean meal Substances 0.000 claims description 12
- 235000012054 meals Nutrition 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 10
- 241000191967 Staphylococcus aureus Species 0.000 claims description 8
- 239000002994 raw material Substances 0.000 claims description 8
- 241000588724 Escherichia coli Species 0.000 claims description 7
- 241000607142 Salmonella Species 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 6
- 239000002773 nucleotide Substances 0.000 claims description 5
- 125000003729 nucleotide group Chemical group 0.000 claims description 5
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 3
- 244000105624 Arachis hypogaea Species 0.000 claims description 3
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 3
- 235000018262 Arachis monticola Nutrition 0.000 claims description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 3
- 235000012343 cottonseed oil Nutrition 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 3
- 235000020232 peanut Nutrition 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 239000011942 biocatalyst Substances 0.000 claims description 2
- 238000004321 preservation Methods 0.000 claims description 2
- 235000000346 sugar Nutrition 0.000 abstract description 21
- 235000013379 molasses Nutrition 0.000 abstract description 7
- 239000001963 growth medium Substances 0.000 description 42
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 32
- 239000007787 solid Substances 0.000 description 25
- 238000010790 dilution Methods 0.000 description 20
- 239000012895 dilution Substances 0.000 description 20
- 230000001580 bacterial effect Effects 0.000 description 16
- 230000003385 bacteriostatic effect Effects 0.000 description 16
- 239000004310 lactic acid Substances 0.000 description 16
- 235000014655 lactic acid Nutrition 0.000 description 16
- 239000007788 liquid Substances 0.000 description 14
- 238000009630 liquid culture Methods 0.000 description 13
- 239000002609 medium Substances 0.000 description 11
- 239000000843 powder Substances 0.000 description 10
- 235000015097 nutrients Nutrition 0.000 description 9
- 241000186660 Lactobacillus Species 0.000 description 8
- 239000003085 diluting agent Substances 0.000 description 8
- 229940039696 lactobacillus Drugs 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- 238000009631 Broth culture Methods 0.000 description 7
- 241001465754 Metazoa Species 0.000 description 7
- 238000012258 culturing Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 6
- 235000010469 Glycine max Nutrition 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 6
- 244000052616 bacterial pathogen Species 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- 108020004465 16S ribosomal RNA Proteins 0.000 description 5
- 239000001888 Peptone Substances 0.000 description 5
- 108010080698 Peptones Proteins 0.000 description 5
- 235000015278 beef Nutrition 0.000 description 5
- 235000019319 peptone Nutrition 0.000 description 5
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 4
- 244000046052 Phaseolus vulgaris Species 0.000 description 4
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 239000011248 coating agent Substances 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 4
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 4
- OQTQHQORDRKHFW-UHFFFAOYSA-L manganese(2+);sulfate;heptahydrate Chemical compound O.O.O.O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O OQTQHQORDRKHFW-UHFFFAOYSA-L 0.000 description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 description 4
- 150000007524 organic acids Chemical class 0.000 description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 4
- 229920000136 polysorbate Polymers 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000001632 sodium acetate Substances 0.000 description 4
- 235000017281 sodium acetate Nutrition 0.000 description 4
- 230000007480 spreading Effects 0.000 description 4
- 238000003892 spreading Methods 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 206010012735 Diarrhoea Diseases 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000433 anti-nutritional effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 230000000529 probiotic effect Effects 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- NOIIUHRQUVNIDD-UHFFFAOYSA-N 3-[[oxo(pyridin-4-yl)methyl]hydrazo]-N-(phenylmethyl)propanamide Chemical compound C=1C=CC=CC=1CNC(=O)CCNNC(=O)C1=CC=NC=C1 NOIIUHRQUVNIDD-UHFFFAOYSA-N 0.000 description 1
- 235000021446 Apple puree Nutrition 0.000 description 1
- 108091005658 Basic proteases Proteins 0.000 description 1
- 235000019750 Crude protein Nutrition 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 235000019730 animal feed additive Nutrition 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 230000005176 gastrointestinal motility Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000006481 glucose medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 235000013613 poultry product Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000004460 silage Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Health & Medical Sciences (AREA)
- Mycology (AREA)
- Botany (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a high temperature and sugar resistant pediococcus acidilactici. Specifically, the invention relates to a new pediococcus acidilactici strain screened from soy molasses, which has the characteristics of high temperature resistance and high sugar resistance. The invention also relates to the application of the novel pediococcus acidilactici.
Description
Technical Field
The present invention relates to the field of microorganisms. More particularly, the invention relates to a new pediococcus acidilactici strain screened from soy molasses, which has the characteristics of high temperature resistance and high sugar resistance. The invention also relates to the application of the novel pediococcus acidilactici.
Background
In recent years, the domestic breeding industry is rapidly developed, so that the feed industry cannot meet the development requirement of the animal husbandry, and a large amount of protein raw materials need to be made up by means of imported fish meal. Therefore, the utilization of plant-derived proteins such as soybean meal, peanut meal and cottonseed meal to replace animal proteins becomes a research and development hotspot. The soybean meal is used as a soybean processing byproduct, the content of crude protein accounts for about 40-50%, and the soybean meal is widely applied to feeds. However, the anti-nutritional factors in the bean pulp are not beneficial to the digestion and absorption of animals and can prevent the growth of the animals. The soybean meal is fermented by using the lactic acid bacteria and other bacterial strains or enzyme preparations, so that the content of anti-nutritional factors in the soybean meal can be effectively reduced, and the utilization rate of nutrition is improved.
On the other hand, the excessive use of antibiotics and other medicines in the past feed and livestock breeding process causes serious medicine residue of livestock and poultry products, and indirectly damages human health. The use of antibiotics in large quantities can also result in the influence on the immunity of animals and be easily infected.
A large number of researches show that the probiotic lactic acid bacteria have good antibacterial property, can be used as an animal feed additive, and have various physiological effects of improving flora, promoting gastrointestinal motility, improving digestibility, improving organism immunity, inhibiting putrefying bacteria and the like. After the feed raw materials are fermented, the lactobacillus grows rapidly, the microorganisms generate heat by breathing, and the temperature of the materials rises. The high-sugar-resistant lactic acid bacteria are beneficial to improving the sugar content in the culture medium during liquid fermentation, optimizing the sugar composition and improving the final bacterial load during high-density culture; during solid fermentation, cheap raw materials such as molasses rich in various sugars and the like are favorably added into the fermentation raw materials, so that the economic benefit is increased.
CN201110419568.3 discloses a high-temperature-resistant and high-sugar-resistant lactic acid bacterium. The mutant lactobacillus rhamnosus can grow well at 55 ℃, and the glucose tolerance concentration can reach 270 g/L.
The high-temperature resistant lactic acid bacterial strain HT1 and the application thereof in the aspect of preparing silage (application number: 201110118135.4) are screened to obtain a strain of lactobacillus rhamnosus which can grow under the condition of 50 ℃.
Screening, identifying and fermenting performance analysis of fruit and vegetable fermented lactobacillus [ J ] food science 2020,41(10):166-171. DOI:10.7506/spkx 1002-6630-20190421-281) from naturally fermented apple puree to obtain 2 strains of lactobacillus, namely lactobacillus plantarum and lactobacillus casei. The two strains are respectively inoculated into MRS culture media with different pH values (pH 2.0-3.0), salt quality fractions (5.0% -10%) or glucose quality fractions (10% -30%) for tolerance verification, and the survival rates are kept above 85%.
Lei Yu et al (Yu L, Pei X, Lei T, et al, Genome shuffle enhanced L-lactic acid production by reconstructing glucose tolerance of Lactobacillus rhamnosus [ J ]. Journal of Biotechnology, 2008, 134(1-2): 154-.
However, there is still a need in the art for probiotic lactic acid bacteria for fermenting feed materials that are better heat and sugar resistant.
Disclosure of Invention
The inventors have screened a new strain of pediococcus acidilactici from soy molasses and have solved the above-mentioned need in the art. The strain has the characteristics of high temperature resistance and high sugar resistance, can be used for fermenting feed raw material products, produces organic acid and increases the palatability of the raw materials. In addition, the strain can inhibit the generation of mixed bacteria in the fermentation process, reduce the risk of mixed bacteria infection in the meal fermentation process and reduce the possibility of causing diarrhea of animals.
In one aspect, the invention relates to a heat-resistant, high-sugar-resistant Pediococcus acidilactici strain, the 16S rDNA of which is shown in SEQ ID NO. 1.
In one embodiment, the pediococcus acidilactici strain is a pediococcus acidilactici strain with a collection number of CGMCC number 21509.
In one embodiment, the pediococcus acidilactici strain is in the form of a live bacterium, a dead bacterium or a cellular fraction thereof.
In one embodiment, the pediococcus acidilactici strain is in an isolated form. The term "isolated" as used herein means separated from its natural environment.
In one embodiment, the strain of pediococcus acidilactici is in a biologically pure form. The term "biologically pure" as used herein refers to a strain in the form of a laboratory culture that is substantially free of other species of organisms. Preferably, the pediococcus acidilactici strain is in the form of a culture of a single biological species.
The term "pediococcus acidilactici strain" as used herein also includes mutant strains of said pediococcus acidilactici strain. The term "mutant" as used herein includes derivative strains comprising a nucleotide sequence having at least 95% identity, preferably at least 96% identity, preferably at least 97% identity, preferably at least 98% identity, more preferably at least 99% identity to SEQ ID No. 1, and comprising mutations in other sequences of the bacterial genome. A mutant may be obtained by genetic engineering techniques, meaning that there is a change in the genetic material of the strain of the invention or that there is recombination of the genetic material of the strain of the invention with other molecules. Typically, to obtain such mutant strains, one skilled in the art may use standard mutagenesis techniques such as UV radiation or exposure to mutagenic chemicals. Sequence "identity" as used herein can be determined using standard techniques known to those skilled in the art. For example, homology can be determined using the online homology algorithm "BLAST" program, which is available at http:// www.ncbi.nlm.nih.gov/BLAST/open.
The term "pediococcus acidilactici strain" as used herein also includes such strains: which comprises a nucleotide sequence which is at least 50, 60, 70, 75, 80, 85 or 90% identical, preferably at least 95%, 96%, 97%, 98% or 99% identical to the nucleotide sequence of SEQ ID NO 1 of the parent strain of Pediococcus acidilactici.
The pediococcus acidilactici strain of the present invention has the characteristics of high temperature resistance and high sugar resistance. In particular, the strains of pediococcus acidilactici of the present invention still have good growth at temperatures of 37-60 ℃, preferably 45-55 ℃, in particular 55 ℃. Furthermore, the Pediococcus acidilactici strain of the present invention can still grow in large amounts at sucrose or glucose concentrations of 300-600g/L, preferably 300-500g/L, especially 500 g/L.
In addition, the pediococcus acidilactici strain of the present invention has a high bacteriostatic effect on various bacteria such as escherichia coli, staphylococcus aureus and salmonella during fermentation.
In another aspect, the present invention relates to a composition comprising the strain of pediococcus acidilactici of the present invention as described above. In one embodiment, the composition is a biocatalyst.
In another aspect, the present invention relates to a method of fermenting a feedstuff comprising adding a pediococcus acidilactici strain of the present invention or a composition of the present invention to a feedstuff and allowing said pediococcus acidilactici strain to ferment said feedstuff to obtain a desired product.
In one embodiment, the process is carried out at a temperature of from 37 to 60 ℃, preferably from 45 to 55 ℃, in particular 55 ℃.
In one embodiment the method is performed at a sucrose or glucose concentration of 300-600g/L, preferably 300-500g/L, in particular 500 g/L.
In one embodiment, the feed raw material is meal, preferably selected from the group consisting of soybean meal, peanut meal and cottonseed meal.
In one embodiment, the method comprises adding the pediococcus acidilactici strain of the present invention or the composition of the present invention, fermenting pulp produces acid under high temperature condition of 55 ℃ and environment of 500g/L sugar concentration, and the pediococcus acidilactici strain inhibits the production of miscellaneous bacteria during the fermentation of the pulp.
In another aspect, the invention also relates to the use of the pediococcus acidilactici strain of the invention for inhibiting undesired bacteria during fermentation.
In one embodiment, the above-mentioned miscellaneous bacteria are selected from the group consisting of Escherichia coli, Salmonella, and Staphylococcus aureus.
Drawings
FIG. 1 shows the growth of Pediococcus acidilactici according to the present invention at various temperatures.
FIG. 2 shows the growth of Pediococcus acidilactici according to the present invention in glucose medium of varying concentration.
FIG. 3 shows the growth of Pediococcus acidilactici according to the invention in sucrose medium at different concentrations.
FIG. 4 shows the bacteriostatic effect of Pediococcus acidilactici on Escherichia coli.
FIG. 5 shows the bacteriostatic effect of Pediococcus acidilactici on Staphylococcus aureus.
FIG. 6 shows the bacteriostatic effect of Pediococcus acidilactici on Salmonella bacteria.
Detailed Description
The inventor selects a new pediococcus acidilactici strain from soy molasses and determines the heat resistance, sugar resistance and bacteriostasis performance of the pediococcus acidilactici strain. The strain is determined to have the characteristics of high temperature resistance and high sugar resistance. The prior art CN201110419568.3 discloses a high-temperature-resistant and high-sugar-resistant lactic acid bacterium, wherein mutant lactobacillus rhamnosus can grow well at 55 ℃, and the glucose tolerance concentration can reach 270 g/L. However, Pediococcus acidilactici according to the present invention is able to tolerate higher glucose concentrations and still grow substantially at sugar concentrations of up to 500g/L, thereby providing lactic acid bacteria that are more capable of meeting the needs of the art. In addition, tests show that the strain can inhibit the generation of mixed bacteria in the fermentation process, reduce the risk of mixed bacteria infection in the meal fermentation process and reduce the possibility of causing diarrhea of animals. 2% of pediococcus acidilactici fermentation liquid is added in the fermentation process of the fermented soybean meal, so that the pH value of the fermented soybean meal can be effectively reduced.
The first embodiment is as follows: screening and identification of bacterial cells
Preparation of culture medium
MRS liquid medium formula: 10g of peptone, 5g of Angel yeast extract, 5g of sodium acetate, 801 g of tween, 0.2g of magnesium sulfate heptahydrate, 10g of beef extract, 20g of glucose, 2g of diammonium citrate, 2g of potassium dihydrogen phosphate, 0.05g of manganese sulfate heptahydrate and 1L of water.
MRS solid medium formula: 1L of MRS liquid culture medium and 1.5-2.0g of agar powder.
The strain of the present invention was isolated from a sample of soy molasses stored by Yihai (defense against urban harbor) Soybean industry Co. The method comprises the following specific steps: approximately 1g of molasses sample was resuspended in 9mL of sterile physiological saline and shaken well. Sucking 1mL of the above liquid, adding into 9mL of sterilized normal saline, and shaking to obtain a dilution of 10 -1 The diluent (2). Repeating the above steps to perform gradient dilution to obtain 10 dilutions -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 The diluent (2). Selection dilution of 10 -5 、10 -6 、10 -7 200 mu L of the diluted solution are respectively coated on MRS solid culture media, and each dilution is divided into two parts. The above operations are carried out under aseptic conditions. After the coating was completed, the cells were cultured in a 37 ℃ incubator. And taking out the plate after 48h, selecting bacterial colonies with the lactobacillus characteristics, such as white, grey white or cream yellow, smooth or slightly rough surface and the like with the diameter of 1-3 mm from the single bacterial colonies on the plate under the aseptic condition, carrying out streak purification on a new MRS solid plate, and culturing for 24h in a constant temperature incubator at 37 ℃. Observing with a microscope to obtain a strain with a spherical shape in microscopic examination. The bacterial solution was used to perform PCR amplification of bacterial 16S rDNA.
The PCR system (50. mu.L) was:
the PCR procedure was: firstly, 5 min at 94 ℃; ② 30 times of circulation at 94 ℃ for 30 s, at 50 ℃ for 30 s and at 72 ℃ for 30 s; ③ 72 ℃ for 5 min.
Sequence of the upstream primer 27F: 5'-AGA GTT TGA TCC TGG CTC AG-3' (SEQ ID NO:2)
The sequence of a downstream primer 1492R: 5 '-GGY TAC CTT GTT ACG ACT T-3' (SEQ ID NO:3)
After the PCR reaction is finished, the PCR product is sent to the biological engineering (Shanghai GmbH) for sequencing after being analyzed and confirmed by nucleic acid electrophoresis. And after a 16S rDNA sequence of the bacteria to be detected is obtained, comparing the sequence with the 16S rDNA sequence recorded in the NCBI website, and separating the non-engineering bacteria wild pediococcus acidilactici.
The sequencing result of the 16S rDNA gene is as follows:
TTAATTGATTATGACGTGCTTGCACTGAATGAGATTTTAACACGAAGTGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCAGAAGCAGGGGATAACACCTGGAAACAGATGCTAATACCGTATAACAGAGAAAACCGCCTGGTTTTCTTTTAAAAGATGGCTCTGCTATCACTTCTGGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGATGATGCGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGTTAAAGAAGAACGTGGGTGAGAGTAACTGTTCACCCAGTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTCTTTTAAGTCTAATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATTGGAAACTGGGAGACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGATTACTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGTAATCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAAGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATCTTCTGCCAACCTAAGAGATTAGGCGTTCCCTTCGGGGACAGAATGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTACTAGTTGCCAGCATTCAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAAACCGCGAGGTTTAGCTAATCTCTTAAAACCATTCTCAGTTCGGACTGTAGGCTGCAACTCGCCTACACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGCCGGTGGGG (SEQ ID NO:1)
and (4) conclusion: the separated strain is pediococcus acidilactici.
The strain is deposited in China general microbiological culture Collection center in 12.21.2020 and classified and named as Pediococcus acidilactici: (Pediococcus acidilactici) The preservation number is CGMCC number 21509.
The second embodiment: determination of the thermostability of Pediococcus acidilactici
1) Preparation of culture medium
MRS liquid medium formula: 10g of peptone, 5g of Angel yeast extract powder, 5g of sodium acetate, 801 g of tween, 0.2g of magnesium sulfate heptahydrate, 10g of beef extract, 20g of glucose, 2g of diammonium citrate, 2g of potassium dihydrogen phosphate, 0.05g of manganese sulfate heptahydrate and 1L of water.
MRS solid medium formula: 1L of MRS liquid culture medium and 1.5-2.0g of agar powder.
2) Inoculating and culturing lactic acid bacteria
In a clean bench, the strain is inoculated in MRS liquid culture medium according to 2% volume percentage, and is respectively subjected to static culture for 14h at 37 ℃, 45 ℃, 50 ℃, 55 ℃ and 60 ℃.
3 viable count
Taking a flat plate with the diameter of about 90 mm, respectively pouring 18-20 mL of the MRS solid culture medium which is heated and melted, uniformly spreading the MRS solid culture medium in the flat plate, and placing the flat plate on a horizontal table to solidify the MRS solid culture medium for later use.
Sucking 1mL of lactobacillus bacterial liquid, adding into 9mL of sterilized normal saline, and oscillating uniformly to obtain the dilution of 10 -1 The diluent (2). Repeating the above procedures to perform gradient dilution to obtain 10 dilutions -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 The diluent (2). Selected dilution of 10 -7 200 mu L of the diluted solution are respectively coated on MRS solid culture medium, and each dilution is divided into two parts. After coating, the cells were incubated at 37 ℃ in a constant temperature incubator. After 48h, the plates were removed and colonies on the plates were counted.
4) Evaluation of Heat resistance
The results are shown in FIG. 1. It can be seen that the pediococcus acidilactici of the present invention still has good growth at 55 ℃ and has heat-resistant characteristics.
Example three: sugar tolerance assay for Pediococcus acidilactici
1) Preparation of culture medium
MRS liquid medium formula: 10g of peptone, 5g of Angel yeast extract powder, 5g of sodium acetate, 801 g of tween, 0.2g of magnesium sulfate heptahydrate, 10g of beef extract, 20g of glucose, 2g of diammonium citrate, 2g of potassium dihydrogen phosphate, 0.05g of manganese sulfate heptahydrate and 1L of water.
MRS solid medium formula: 1L of MRS liquid culture medium and 1.5-2.0g of agar powder.
The glucose in the solution is replaced by 100g/L glucose, 200g/L glucose, 300g/L glucose, 400g/L glucose, 500g/L glucose, 600g/L glucose, 100g/L sucrose, 200g/L sucrose, 300g/L sucrose, 400g/L sucrose, 500g/L sucrose and 600g/L sucrose respectively.
2) Inoculating and culturing lactic acid bacteria
In a clean bench, the strain is respectively inoculated in MRS liquid culture medium, 100g/L glucose, 200g/L glucose, 300g/L glucose, 400g/L glucose, 500g/L glucose, 600g/L glucose, 100g/L sucrose, 200g/L sucrose, 300g/L sucrose, 400g/L sucrose, 500g/L sucrose, 600g/L sucrose liquid culture medium according to the inoculation amount of 2 percent, and is statically cultured for 14 hours at 37 ℃.
3) Viable count
Taking a flat plate with the diameter of about 90 mm, respectively pouring 18-20 mL of the MRS solid culture medium which is heated and melted, uniformly spreading the MRS solid culture medium in the flat plate, and placing the flat plate on a horizontal table to solidify the MRS solid culture medium for later use.
Sucking 1mL of lactobacillus bacterial liquid, adding into 9mL of sterilized normal saline, and oscillating uniformly to obtain the dilution of 10 -1 The diluent (2). Repeating the above steps to perform gradient dilution to obtain 10 dilutions -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 The diluent (2). Selected dilution of 10 -5 、10 -6 、10 -7 200 mu L of the diluted solution are respectively coated on MRS solid culture media, and each dilution is divided into two parts. After the coating was completed, the cells were cultured in a 37 ℃ incubator. After 48h, the plates were removed and colonies on the plates were counted.
4) Evaluation results of sugar resistance
The results are shown in FIGS. 2-3. As can be seen from the figure, the isolated pediococcus acidilactici medium grows well, still grows a lot at a sugar concentration of 500g/L, and has the characteristic of high sugar resistance.
Example four: growth of Pediococcus acidilactici in high temperature and high sugar environment
1) Preparation of culture medium
MRS liquid medium formula: 10g of peptone, 5g of Angel yeast extract powder, 5g of sodium acetate, 801 g of tween, 0.2g of magnesium sulfate heptahydrate, 10g of beef extract, 20g of glucose, 2g of diammonium citrate, 2g of potassium dihydrogen phosphate, 0.05g of manganese sulfate heptahydrate and 1L of water.
MRS solid medium formula: 1L of MRS liquid culture medium and 1.5-2.0g of agar powder.
The glucose in the solution is replaced by 500g/L glucose and 500g/L sucrose respectively.
2) Inoculating and culturing lactic acid bacteria
In a clean bench, the strain is respectively inoculated in MRS liquid culture medium, 500g/L glucose and 500g/L sucrose liquid culture medium according to the inoculation amount of 2%, the MRS liquid culture medium is statically cultured for 14h at 37 ℃, and the 500g/L glucose and 500g/L sucrose liquid culture medium is statically cultured for 14h at 55 ℃.
3) Viable count
Taking a flat plate with the diameter of about 90 mm, respectively pouring 18-20 mL of the MRS solid culture medium which is heated and melted, uniformly spreading the MRS solid culture medium in the flat plate, and placing the flat plate on a horizontal table to solidify the MRS solid culture medium for later use.
Sucking 1mL of lactobacillus bacterial liquid, adding into 9mL of sterilized normal saline, and oscillating uniformly to obtain the dilution of 10 -1 The diluent of (4). Repeating the above procedures to perform gradient dilution to obtain 10 dilutions -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 The diluent of (4). Selected dilution of 10 -5 、10 -6 、10 -7 200 mu L of the diluted solution are respectively coated on MRS solid culture media, and each dilution is divided into two parts. After coating, the cells were incubated at 37 ℃ in a constant temperature incubator. After 48h, the plates were removed and colonies on the plates were counted.
4) Growth of Pediococcus acidilactici
The results are shown in the table, and the table shows that a large amount of pediococcus acidilactici can grow at the sugar concentration of 500g/L at the temperature of 55 ℃, and has better high temperature resistance and high sugar resistance.
Example five: method for detecting bacteriostatic activity of pediococcus acidilactici by Oxford cup method
1) Preparation of culture medium
The formula of the liquid nutrient broth culture medium comprises the following components: 10g of peptone, 3g of beef powder, 5g of sodium chloride and 1L of water;
solid nutrient broth culture medium formula: 1L of nutrient broth liquid culture medium and 2.0g of agar powder;
semi-solid nutrient broth culture medium formula: 1L of nutrient broth liquid culture medium and 1.0 g of agar powder.
2) Activated pathogenic bacteria
Inoculating the glycerol tube of pathogenic bacteria such as Escherichia coli, salmonella and Staphylococcus aureus into liquid nutrient broth culture medium according to 2% volume percentage, and culturing for 16 hours at 37 ℃ by shaking table 200 r/min.
3) Preparation of pediococcus acidilactici bacterial suspension
In a clean bench, the strain is inoculated in 100 mL of MRS liquid medium and is statically cultured for 24h at 37 ℃.
4) Preparation of a double-layer plate
Taking a flat plate with the diameter of about 90 mm, respectively pouring 20 mL of the solid nutrient broth culture medium which is heated and melted, uniformly spreading the solid nutrient broth culture medium in the flat plate, and placing the flat plate on a horizontal table to solidify the solid nutrient broth culture medium to be used as a bottom layer. Heating and melting a proper amount of semi-solid nutrient agar culture medium, cooling to 50 ℃, adding 0.1-0.2 mL of pathogenic bacteria suspension into each 50-100 mL of culture medium, and adding 5 mL of pathogenic bacteria suspension into each plate respectively to uniformly spread the pathogenic bacteria suspension on the bottom layer to serve as a bacteria layer. After being placed on a horizontal table surface for cooling, 3 oxford cups are uniformly arranged in each flat plate at equal intervals (about 4 cm) for later use.
5) Bacteriostatic test
The prepared pediococcus acidilactici bacterial suspension is taken, 200 mu L of the prepared pediococcus acidilactici bacterial suspension is respectively dripped into each oxford cup in each double-layer flat plate, and after the oxford cups are cultured for 18 hours at 37 ℃, the diameter (or the area) of each inhibition zone is measured to make evaluation.
Remarking: 1) the oxford cup must be placed stably and in place to ensure that the bottom is accurately inserted between two layers of flat plates; 2) when the antibacterial substance is added, the antibacterial substance is completely added into the oxford cup and is not sprayed on the outer wall or the outer part of the oxford cup, and if the antibacterial substance is sprayed outside, the growth of lactic acid bacteria in an antibacterial zone can be formed; 3) if the growth of the lactic acid bacteria outside the Oxford cup can not be controlled, the lactic acid bacteria can be centrifuged in a sterile centrifuge tube, and supernatant of the lactic acid bacteria can be taken and added into the Oxford cup for bacteriostatic experiments.
6) Evaluation results of antibacterial Properties
Bacteriostasis: the bacteriostatic action is no longer than 10 mm, moderate bacteriostatic action is taken when the bacteriostatic zone is less than 10 mm and is less than 15 mm, and high bacteriostatic action is taken when the bacteriostatic zone is more than 15 mm.
Remarking: 1) the larger the inhibition zone is, the better the inhibition effect is; 2) the bacteriostatic circle refers to the whole diameter of the circular transparent circle; 3) the external diameter of the oxford cup is 8 mm.
The bacteriostatic effect is shown in figures 4-6, and the specific measurement values are as follows:
| pathogenic bacteria | Diameter of bacteriostatic circle (mm) |
| Escherichia coli | 23 |
| Staphylococcus aureus (Staphylococcus aureus) | 22.99 |
| Salmonella | 24.29 |
And (4) conclusion:
the separated pediococcus acidilactici has high bacteriostatic action on escherichia coli, staphylococcus aureus and salmonella.
Example six: preparation of fermented soybean meal
50g of bean pulp is put into a self-sealing bag, alkaline protease (purchased from New Yangshao bio-enzyme preparation Co., Ltd., Henan) is added according to the proportion of 100 plus 500U/g of the bean pulp, the mixture is uniformly mixed, and water and lactobacillus accounting for 2 percent of the mass of the bean pulp are added to ensure that the water content of the mixed sample is about 37 percent. And mixing uniformly again, putting the sample into a constant-temperature incubator at 40 ℃ for culturing for 72h, drying in an oven at 65 ℃ for 6h after culturing for 72h, crushing, sieving by a 60-mesh sieve, and measuring the pH value and the organic acid content by adopting a titration method.
| Group of | Lactic acid bacteria composition | pH | Titration of organic acid content% (10% moisture) |
| Bacterial liquid blank group | —— | 6.08 | 2.06 |
| Pediococcus acidilactici group | Pediococcus acidilactici | 4.45 | 3.68 |
And (4) conclusion: 2% of pediococcus acidilactici fermentation liquor is added in the fermentation process of the fermented soybean meal, the pH value of the fermented soybean meal is effectively reduced to 4.45, the content of organic acid is effectively improved, and the digestion and absorption of animals to feed are facilitated.
Sequence listing
<110> Fengyi (Shanghai) Biotechnology research and development center, Inc
<120> high temperature resistant and high sugar resistant Pediococcus acidilactici
<130> CPCH2062876N
<160> 3
<170> PatentIn version 3.5
<210> 1
<211> 1402
<212> DNA
<213> Pediococcus acidilactici
<400> 1
ttaattgatt atgacgtgct tgcactgaat gagattttaa cacgaagtga gtggcggacg 60
ggtgagtaac acgtgggtaa cctgcccaga agcaggggat aacacctgga aacagatgct 120
aataccgtat aacagagaaa accgcctggt tttcttttaa aagatggctc tgctatcact 180
tctggatgga cccgcggcgc attagctagt tggtgaggta acggctcacc aaggcgatga 240
tgcgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg cccagactcc 300
tacgggaggc agcagtaggg aatcttccac aatggacgca agtctgatgg agcaacgccg 360
cgtgagtgaa gaagggtttc ggctcgtaaa gctctgttgt taaagaagaa cgtgggtgag 420
agtaactgtt cacccagtga cggtatttaa ccagaaagcc acggctaact acgtgccagc 480
agccgcggta atacgtaggt ggcaagcgtt atccggattt attgggcgta aagcgagcgc 540
aggcggtctt ttaagtctaa tgtgaaagcc ttcggctcaa ccgaagaagt gcattggaaa 600
ctgggagact tgagtgcaga agaggacagt ggaactccat gtgtagcggt gaaatgcgta 660
gatatatgga agaacaccag tggcgaaggc ggctgtctgg tctgtaactg acgctgaggc 720
tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg taaacgatga 780
ttactaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt aagtaatccg 840
cctggggagt acgaccgcaa ggttgaaact caaaagaatt gacgggggcc cgcacaagcg 900
gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct tgacatcttc 960
tgccaaccta agagattagg cgttcccttc ggggacagaa tgacaggtgg tgcatggttg 1020
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa cccttattac 1080
tagttgccag cattcagttg ggcactctag tgagactgcc ggtgacaaac cggaggaagg 1140
tggggacgac gtcaaatcat catgcccctt atgacctggg ctacacacgt gctacaatgg 1200
atggtacaac gagttgcgaa accgcgaggt ttagctaatc tcttaaaacc attctcagtt 1260
cggactgtag gctgcaactc gcctacacga agtcggaatc gctagtaatc gcggatcagc 1320
atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc atgagagttt 1380
gtaacaccca aagccggtgg gg 1402
<210> 2
<211> 20
<212> DNA
<213> Artificial sequence
<220>
<223> primer
<400> 2
<210> 3
<211> 19
<212> DNA
<213> Artificial sequence
<220>
<223> primer
<400> 3
ggytaccttg ttacgactt 19
Claims (10)
1. A thermotolerant, high-sugar tolerant pediococcus acidilactici strain comprising a nucleotide sequence having at least 95% identity, preferably at least 99% identity with SEQ ID No. 1, most preferably it comprises the nucleotide sequence of SEQ ID No. 1.
2. The pediococcus acidilactici strain of claim 1, which is a pediococcus acidilactici strain with a preservation number of CGMCC number 21509.
3. The pediococcus acidilactici strain of claim 1 which is a recombinant strain.
4. A composition comprising the pediococcus acidilactici strain of any one of claims 1-3.
5. The composition of claim 4 which is a biocatalyst.
6. A method of fermenting a feedstuff comprising adding the pediococcus acidilactici strain of any one of claims 1-3 or the composition of claim 4 or 5 to a feedstuff and allowing the pediococcus acidilactici strain to ferment the feedstuff to obtain a desired product.
7. The process according to claim 6, wherein the process is carried out at a temperature of 37 to 60 ℃, preferably 45 to 55 ℃, in particular 55 ℃.
8. The process of claim 6 or 7, wherein the process is carried out at a sucrose or glucose concentration of 300-600g/L, preferably 300-500g/L, in particular 500 g/L.
9. The method according to any one of claims 6-8, wherein the feed raw material is meal, preferably selected from the group consisting of soybean meal, peanut meal and cottonseed meal.
10. Use of the pediococcus acidilactici strain of any of claims 1-3 for inhibiting during fermentation undesired bacteria, preferably undesired bacteria selected from the group consisting of escherichia coli, salmonella, and staphylococcus aureus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110245170.6A CN115011498A (en) | 2021-03-05 | 2021-03-05 | High-temperature-resistant and high-sugar-resistant pediococcus acidilactici |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110245170.6A CN115011498A (en) | 2021-03-05 | 2021-03-05 | High-temperature-resistant and high-sugar-resistant pediococcus acidilactici |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115011498A true CN115011498A (en) | 2022-09-06 |
Family
ID=83064690
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110245170.6A Pending CN115011498A (en) | 2021-03-05 | 2021-03-05 | High-temperature-resistant and high-sugar-resistant pediococcus acidilactici |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115011498A (en) |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102533591A (en) * | 2011-12-15 | 2012-07-04 | 天津工业大学 | High temperature resisting and high-glucose resisting lactic acid bacteria |
| CN102586348A (en) * | 2011-01-14 | 2012-07-18 | 华东理工大学 | Preparation method of lactic acid by saccharifying and fermenting lignocellulose |
| CN103103224A (en) * | 2011-11-10 | 2013-05-15 | 华东理工大学 | Method for producing high-concentration lactic acid by lignocellulose at high temperature |
| US20160193259A1 (en) * | 2011-11-14 | 2016-07-07 | Imagilin Technology, LLC. | High temperature resistant probiotics for food and feed preparations |
| CN105925499A (en) * | 2016-04-29 | 2016-09-07 | 四川农业大学 | Pediococcus acidilactici JQQ2 and applications thereof |
| JP5988455B2 (en) * | 2013-01-11 | 2016-09-07 | 株式会社タカギ | New lactic acid bacteria |
| CN108504601A (en) * | 2018-04-10 | 2018-09-07 | 北京好实沃生物技术有限公司 | One plant of Pediococcus acidilactici HEW-AP27 and its application |
| CN109536409A (en) * | 2018-12-17 | 2019-03-29 | 吉林中粮生化有限公司 | A kind of resistance is high and using the Pedicoccus acidilacticii strain of several kinds of carbon source and the method for producing lactic acid using the bacterial strain |
| 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 |
| CN110373361A (en) * | 2019-08-12 | 2019-10-25 | 日照东润德农牧发展有限公司 | A kind of Pediococcus acidilactici micro-capsule and preparation method thereof |
| CN111793586A (en) * | 2020-08-13 | 2020-10-20 | 石河子大学 | Breast milk source pediococcus pentosaceus and application thereof |
| CN115011497A (en) * | 2021-03-05 | 2022-09-06 | 丰益(上海)生物技术研发中心有限公司 | Pediococcus acidilactici and application thereof |
-
2021
- 2021-03-05 CN CN202110245170.6A patent/CN115011498A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586348A (en) * | 2011-01-14 | 2012-07-18 | 华东理工大学 | Preparation method of lactic acid by saccharifying and fermenting lignocellulose |
| CN103103224A (en) * | 2011-11-10 | 2013-05-15 | 华东理工大学 | Method for producing high-concentration lactic acid by lignocellulose at high temperature |
| US20160193259A1 (en) * | 2011-11-14 | 2016-07-07 | Imagilin Technology, LLC. | High temperature resistant probiotics for food and feed preparations |
| CN102533591A (en) * | 2011-12-15 | 2012-07-04 | 天津工业大学 | High temperature resisting and high-glucose resisting lactic acid bacteria |
| JP5988455B2 (en) * | 2013-01-11 | 2016-09-07 | 株式会社タカギ | New lactic acid bacteria |
| CN105925499A (en) * | 2016-04-29 | 2016-09-07 | 四川农业大学 | Pediococcus acidilactici JQQ2 and applications thereof |
| CN108504601A (en) * | 2018-04-10 | 2018-09-07 | 北京好实沃生物技术有限公司 | One plant of Pediococcus acidilactici HEW-AP27 and its application |
| CN109536409A (en) * | 2018-12-17 | 2019-03-29 | 吉林中粮生化有限公司 | A kind of resistance is high and using the Pedicoccus acidilacticii strain of several kinds of carbon source and the method for producing lactic acid using the bacterial strain |
| 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 |
| CN110373361A (en) * | 2019-08-12 | 2019-10-25 | 日照东润德农牧发展有限公司 | A kind of Pediococcus acidilactici micro-capsule and preparation method thereof |
| CN111793586A (en) * | 2020-08-13 | 2020-10-20 | 石河子大学 | Breast milk source pediococcus pentosaceus and application thereof |
| CN115011497A (en) * | 2021-03-05 | 2022-09-06 | 丰益(上海)生物技术研发中心有限公司 | Pediococcus acidilactici and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| BANG J 等: "15 of 172 Heat Tolerances of Salmonella, Cronobacter sakazakii, and Pediococcus acidilactici Inoculated into Galactooligosaccharide", JOURNAL OF FOOD PROTECTION, vol. 80, no. 7, 1 July 2017 (2017-07-01), pages 1123 - 1127 * |
| KAI ZHAO 等: "Simultaneous saccharification and high titer lactic acid fermentation of corn stover using a newly isolated lactic acid bacterium Pediococcus acidilactici DQ2", BIORESOUR TECHNOL, vol. 135, 27 September 2012 (2012-09-27), pages 481 - 489 * |
| 楚德强 等: "一株高耐受性乳酸菌的分离及其在木质纤维素发酵生产高浓度L-乳酸中的应用", 华东理工大学学报(自然科学版), vol. 37, no. 5, 15 October 2011 (2011-10-15), pages 521 - 523 * |
| 董婷 等: "乳酸片球菌PA003抑菌作用及体外耐受性的研究", 食品工业科技, no. 5, 8 November 2013 (2013-11-08), pages 140 - 144 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108587957B (en) | Screening and application of probiotic bacillus licheniformis capable of highly producing complex enzyme | |
| CN110591938B (en) | High-stress-resistance bacillus coagulans mutant strain and application thereof | |
| CN113151064B (en) | Pediococcus pentosaceus and application thereof | |
| CN113080309B (en) | Bacillus subtilis with antibacterial and antiviral functions and application thereof | |
| CN110791460B (en) | Strain for inhibiting growth of aspergillus flavus and generation of toxin | |
| CN116179440B (en) | Bacillus gallinarum and application thereof | |
| CN115094012B (en) | Preparation method and application of bacillus coagulans BC-HYC strain microbial inoculum | |
| CN113604404B (en) | Bacillus coagulans YSF17 and application thereof | |
| CN112980735B (en) | Clostridium butyricum, microbial inoculum, application of clostridium butyricum and microbial inoculum and preparation method of microbial inoculum | |
| CN110257302B (en) | Screening method and application of lactobacillus strain with antioxidant capacity | |
| KR100541379B1 (en) | Animal feed additives and method of producing the same | |
| CN114806975A (en) | Microecological preparation containing enterogenous probiotics as well as preparation method and application of microecological preparation | |
| CN108179122A (en) | A kind of probiotic enterococcus faecium of high adherency and its application | |
| CN108546663B (en) | Porcine lactobacillus crispatus and application thereof | |
| CN111996139B (en) | Composite microecological preparation for improving production benefit of meat calves and application thereof | |
| CN112226389A (en) | Planting culture method of intestinal probiotic groups of Sanhuang young chickens and application of intestinal probiotic groups | |
| CN109423467B (en) | Lactobacillus plantarum and application thereof in fields of food and feed | |
| CN105002123B (en) | A kind of preparation method of the bacillus coagulans Liu-g1 active bacteria formulations of production neutral proteinase | |
| CN114908017B (en) | Lactobacillus plantarum and application thereof in silage | |
| KR20070071911A (en) | Novel lactobacillus sakei and use thereof | |
| KR101431250B1 (en) | Poultry fermented additive feed composition containing the novel Lactic acid bacteria having anit-pathogenic microorganism and Organic matter decomposition activity microorganism | |
| CN104651270B (en) | Feed probiotic agent prepared by one plant of pig source bacillus subtilis and its solid state fermentation | |
| CN114181867B (en) | Lactobacillus plantarum, and product and application thereof | |
| CN116286514A (en) | Multifunctional microbial inoculum, lactobacillus reuteri contained therein and application thereof | |
| CN115011498A (en) | High-temperature-resistant and high-sugar-resistant pediococcus acidilactici |
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 |