CN110122577B - Method for preparing fermented soybean milk from paenibacillus bovis, prepared fermented soybean milk and application - Google Patents
Method for preparing fermented soybean milk from paenibacillus bovis, prepared fermented soybean milk and application Download PDFInfo
- Publication number
- CN110122577B CN110122577B CN201910435092.9A CN201910435092A CN110122577B CN 110122577 B CN110122577 B CN 110122577B CN 201910435092 A CN201910435092 A CN 201910435092A CN 110122577 B CN110122577 B CN 110122577B
- Authority
- CN
- China
- Prior art keywords
- soybean milk
- fermented soybean
- streptococcus mutans
- fermentation
- formation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 195
- 244000068988 Glycine max Species 0.000 title claims abstract description 195
- 235000013336 milk Nutrition 0.000 title claims abstract description 195
- 210000004080 milk Anatomy 0.000 title claims abstract description 195
- 239000008267 milk Substances 0.000 title claims abstract description 193
- 241000201493 Paenibacillus bovis Species 0.000 title claims abstract description 80
- 238000000034 method Methods 0.000 title claims abstract description 48
- 241000194019 Streptococcus mutans Species 0.000 claims abstract description 98
- 238000000855 fermentation Methods 0.000 claims abstract description 57
- 230000004151 fermentation Effects 0.000 claims abstract description 57
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 47
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 5
- 230000005764 inhibitory process Effects 0.000 claims description 46
- 238000002360 preparation method Methods 0.000 claims description 39
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 31
- 239000007787 solid Substances 0.000 claims description 30
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 29
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 29
- 238000011081 inoculation Methods 0.000 claims description 26
- 244000046052 Phaseolus vulgaris Species 0.000 claims description 15
- 239000002054 inoculum Substances 0.000 claims description 9
- 240000004922 Vigna radiata Species 0.000 claims description 7
- 235000010721 Vigna radiata var radiata Nutrition 0.000 claims description 7
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 claims description 7
- 230000003068 static effect Effects 0.000 claims description 7
- 240000006677 Vicia faba Species 0.000 claims description 6
- 235000010749 Vicia faba Nutrition 0.000 claims description 6
- 235000002098 Vicia faba var. major Nutrition 0.000 claims description 6
- 235000011453 Vigna umbellata Nutrition 0.000 claims description 6
- 240000001417 Vigna umbellata Species 0.000 claims description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 44
- 239000001963 growth medium Substances 0.000 abstract description 22
- 230000000694 effects Effects 0.000 abstract description 19
- 208000002925 dental caries Diseases 0.000 abstract description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 30
- 239000000523 sample Substances 0.000 description 24
- 230000032770 biofilm formation Effects 0.000 description 22
- 239000012153 distilled water Substances 0.000 description 14
- 239000000047 product Substances 0.000 description 14
- 230000001580 bacterial effect Effects 0.000 description 13
- 239000007788 liquid Substances 0.000 description 12
- 238000009630 liquid culture Methods 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 11
- 239000000725 suspension Substances 0.000 description 11
- 239000000463 material Substances 0.000 description 10
- 238000012360 testing method Methods 0.000 description 8
- 235000013322 soy milk Nutrition 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 201000010099 disease Diseases 0.000 description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 244000199866 Lactobacillus casei Species 0.000 description 5
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 5
- 241000194020 Streptococcus thermophilus Species 0.000 description 5
- 241001052560 Thallis Species 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 238000002791 soaking Methods 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 235000019640 taste Nutrition 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- 235000013958 Lactobacillus casei Nutrition 0.000 description 4
- 229940017800 lactobacillus casei Drugs 0.000 description 4
- 241000282414 Homo sapiens Species 0.000 description 3
- 241000194024 Streptococcus salivarius Species 0.000 description 3
- 230000001013 cariogenic effect Effects 0.000 description 3
- 239000008176 lyophilized powder Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 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 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000002068 microbial inoculum Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000000214 mouth Anatomy 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000005057 refrigeration Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 241000186046 Actinomyces Species 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 201000004328 Pulpitis Diseases 0.000 description 1
- 206010037464 Pulpitis dental Diseases 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 208000008312 Tooth Loss Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002053 acidogenic effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000005115 demineralization Methods 0.000 description 1
- 230000002328 demineralizing effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 235000021332 kidney beans Nutrition 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 208000004480 periapical periodontitis Diseases 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 229910001961 silver nitrate Inorganic materials 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000011146 sterile filtration Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000012134 supernatant fraction Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
-
- 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
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Beans For Foods Or Fodder (AREA)
Abstract
The invention discloses a method for preparing fermented soybean milk by using paenibacillus bovis, which comprises the following steps: the Paenibacillus bovis CGMCC No.8333 is used as a zymophyte agent to be inoculated into the soybean milk for fermentation, so as to obtain the fermented soybean milk. The fermented soybean milk prepared by the method can obviously reduce the formation amount of streptococcus mutans biomembranes, and the novel application of the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 fermented soybean milk in resisting dental caries is disclosed, and the fermented soybean milk has a good application prospect. Compared with other strains capable of fermenting soybean milk or culture media in which the strains can grow and reproduce, the fermented soybean milk prepared by the CGMCC No.8333 has obvious activity of inhibiting the formation of streptococcus mutans biofilms.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a preparation method of fermented soybean milk. In addition, the invention also relates to fermented soybean milk and application thereof.
Background
Dental caries, commonly known as dental caries and decayed tooth, is a bacterial disease which can cause secondary pulpitis and periapical periodontitis, and even inflammation of alveolar bone and jaw bone. If not treated in time, the lesion continues to develop, forming a cavity, eventually the crown is completely destroyed and disappears, the end result of which is the loss of teeth. Dental caries is characterized by high incidence rate and wide distribution. The disease is a main common disease in the oral cavity and is one of the most common diseases of human beings, and the world health organization combines the disease with tumor and cardiovascular diseases as three major important diseases for preventing and treating the diseases of human beings.
There are many factors that cause dental caries, such as bacteria, oral environment (food, saliva), host, etc., wherein the bacteria are necessary conditions for dental caries, and it is generally considered that there are two types of cariogenic bacteria, one of which is acidogenic bacteria, mainly Streptococcus mutans (s.mutans), actinomyces and lactobacillus, which decompose carbohydrate to produce acid, resulting in inorganic demineralization of teeth; the other is gram-positive coccus, which can destroy organic matters and can form caries in teeth after long-term action. Among the above cariogenic bacteria, Streptococcus mutans is recognized by scholars at home and abroad as the most important and major pathogenic bacterium for caries. However, these pathogenic bacteria do not directly colonize the tooth surface, but indirectly adhere to the tooth surface after forming plaque by binding with biofilm (insoluble polysaccharide) produced by streptococcus mutans, and the degree of caries gradually worsens as cariogenic bacteria accumulate on the biofilm. Therefore, the aim of preventing and treating the decayed tooth can be achieved by reducing the yield of the biological film.
At present, most of the reports in the research on the treatment of the dental caries focus on controlling or reducing the quantity of streptococcus mutans in the oral cavity, however, the healthy oral environment is generally in a state that harmful bacteria and beneficial bacteria reach a balance, and excessive reduction of the harmful bacteria can bring about imbalance of oral flora, so as to cause other adverse reactions of human bodies. In addition, some clinical bactericidal drugs against streptococcus mutans, such as ammonia, silver nitrate and other compounds, have strong side effects, stain teeth after long-term use, and are not suitable for anterior tooth treatment. The risk of caries can also be reduced by reducing the yield of streptococcus mutans biofilm, and the inherent balance of oral flora is not destroyed or side effects are generated, but the reports in the aspect are relatively less, especially the research related to microbial fermentation products is extremely limited, and the fermentation products for inhibiting the formation of the streptococcus mutans biofilm are relatively short.
Therefore, it is urgent to provide a fermented product having an effect of inhibiting the formation of a streptococcus mutans biofilm. This is a problem that those skilled in the art are at risk for solving.
Disclosure of Invention
Based on the technical problems, the invention provides a method for preparing fermented soybean milk, and the fermented soybean milk with the effect of inhibiting the formation of streptococcus mutans biofilm can be prepared by the method.
Specifically, on the one hand, the preparation method of the fermented soybean milk is provided, and comprises the following steps: the Paenibacillus bovis CGMCC No.8333 is used as a zymophyte agent to be inoculated into the soybean milk for fermentation, so as to obtain the fermented soybean milk.
The Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 strain is preserved in China general microbiological culture Collection center (CGMCC) in 2013, 10 months and 14 days, and the preservation address is as follows: west road No.1, north chen, chaoyang district, beijing, zip code: 100101. the preservation information of the strain of Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 is disclosed in patent CN 103740618B.
Further, the inoculation amount of the zymophyte agent is 2x10 7 ～1x10 8 cfu/mL。
The fermentation inoculum also comprises lactobacillus plantarum ATCC14917, and the inoculation ratio of the lactobacillus plantarum ATCC14917 to paenibacillus bovis CGMCC No.8333 is 1-9: 1 to 9.
When lactobacillus plantarum ATCC14917 is purchased from ATCC, lactobacillus plantarum ATCC14917 and paenibacillus bovis CGMCC No.8333 are used as the composite microbial inoculum to ferment soybean milk, the prepared fermented soybean milk has obvious activity of inhibiting the formation of streptococcus mutans biofilm.
Further, the soybean milk is at least one of soybean, red bean, mung bean, broad bean, white kidney bean or small white kidney bean soybean milk.
Further, the mass percentage content of solid matters in the soybean milk is 1-9%.
Further, the fermentation temperature is 15-45 ℃, and the fermentation time is 2-48 h.
Further, the fermentation is static culture.
In a second aspect, there is also provided a fermented soybean milk prepared by any one of the above-mentioned preparation methods.
Furthermore, the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm is more than or equal to 46 percent.
In a third aspect, the application of the fermented soybean milk in preparing the anti-dental caries food is also provided.
Compared with the prior art, the preparation method in the technical scheme adopts Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 for the first time, and prepares the fermented soybean milk with the streptococcus mutans biofilm inhibiting activity by fermenting the soybean milk as a culture medium, thereby disclosing the new application of the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 fermented soybean milk in resisting decayed teeth. Compared with other strains capable of fermenting soybean milk or culture media in which the strains can grow and reproduce, the fermented soybean milk prepared by the CGMCC No.8333 has obvious activity of inhibiting the formation of streptococcus mutans biofilms.
Detailed Description
In order to more clearly illustrate the technical solution of the present invention, the technical solution of the present invention will be further illustrated with reference to the following specific embodiments:
in a specific embodiment, there is provided a method for preparing fermented soybean milk, comprising the steps of: and inoculating the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 into the soybean milk for fermentation to obtain the fermented soybean milk.
According to the preparation method in the technical scheme, the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 is used as a fermentation microbial agent for the first time, the soybean milk is used as a culture medium for fermentation, the fermented soybean milk with the streptococcus mutans biofilm inhibition activity is prepared, the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 has the streptococcus mutans inhibition activity by utilizing metabolic substances synthesized by the soybean milk, and the novel application of the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 fermented soybean milk in resisting dental caries is disclosed. Compared with other strains capable of fermenting soybean milk or culture media in which the strains can grow and reproduce, the fermented soybean milk prepared by the CGMCC No.8333 has more remarkable inhibiting effect on the formation of streptococcus mutans biofilm and has good stability.
Further, in the above step, the inoculation amount of the fermentation inoculum is 2x10 7 ～1x10 8 cfu/mL; preferably 4x10 7 ～8x10 7 cfu/mL, more preferably 6X10 7 cfu/mL。
The fermentation inoculum also comprises lactobacillus plantarum ATCC14917, wherein the inoculation ratio of the lactobacillus plantarum ATCC14917 to the paenibacillus bovis CGMCC No.8333 is 1-9: 1 to 9. When lactobacillus plantarum ATCC14917 and paenibacillus bovis CGMCC No.8333 are used as the composite microbial inoculum to ferment the soybean milk, the prepared fermented soybean milk has remarkable activity of inhibiting the formation of streptococcus mutans biofilms.
Further, in the above step, the soybean milk is at least one of soybean, red bean, mung bean, broad bean, white kidney bean and small white kidney bean milk, and more preferably soybean milk.
Further, in the step, the mass percentage content of the solid matters in the soybean milk is 1-9%; preferably 3% -7%; more preferably 5%. The content of solid matters in the soybean milk has a direct relation with the biofilm inhibition effect of a final product, when the content of the solid matters is too high, the osmotic pressure of the environment of thalli is increased, the growth of the thalli is influenced, the synthetic amount of the biofilm inhibition substances is reduced, in addition, the production cost is increased, and when the content of the solid matters is too low, the proliferation effect of viable bacteria is poor due to insufficient nutrient components, and the generation of the biofilm inhibition substances is also influenced.
Further, in the above step, the preferable fermentation temperature is 15 ℃ to 45 ℃; preferably 25 ℃ to 40 ℃; more preferably 37 deg.c.
Further, in the step, the preferable fermentation time is 2-48 h; preferably 12 to 36 hours; more preferably 24 h.
In the above step, the preferred fermentation method is static culture. Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 is facultative anaerobe, and can grow and propagate rapidly under the condition of static culture.
It is also understood from the combination of example and comparative example 1 that the inhibitory effect of fermented soybean milk prepared from Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 on the biofilm formation of Streptococcus mutans is significantly reduced when the preferable fermentation parameters are outside the range. In the preferable range, the inoculation amount, the soybean milk concentration, the culture temperature and the fermentation time are mutually influenced, so that the fermented soybean milk prepared by the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 has better inhibiting effect on the formation of the streptococcus mutans biofilm.
In another embodiment, there is provided a fermented soybean milk produced by any one of the above-mentioned methods for producing fermented soybean milk.
Since the preparation method of the fermented soybean milk has the above-mentioned advantageous effects, the fermented soybean milk prepared by the preparation method also has corresponding advantageous effects, which are not described herein again.
Furthermore, the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm is more than or equal to 46 percent. The combination of the examples shows that the inhibition effect of the fermented soybean milk on the formation of the streptococcus mutans biofilm is obviously higher than that of other strains of the fermentable soybean milk.
The following examples further illustrate the above embodiments, but do not therefore limit the invention within the scope of the examples described. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.
In the examples below, all the starting materials are commercially available and meet the relevant national standards.
Example 1
1. Materials and methods
(a) Preparation of Paenibacillus bovis seeds (fermentation strain): dissolving lyophilized powder of Bacillus bovis CGMCC No.8333 with small amount of sterile distilled water, taking out an inoculating loop, streaking on TYC solid culture medium (purchased from OXOID Co., UK), aerobically culturing at 30 deg.C for 48h, taking out, selecting single colony with inoculating loop, placing into 10mL TYC liquid culture medium (purchased from OXOID Co., UK), uniformly dispersing the colony in the liquid culture medium by vortex oscillator, shake-culturing at 30 deg.C and 180rpm for 24h, taking out, inoculating into TYC liquid culture medium (purchased from OXOID Co., UK) with 2% (v/v) inoculum size, shake-culturing at 30 deg.C and 180rpm for 24h, centrifuging at 15000rpm for 10min, discarding supernatant, washing thallus with sterile distilled water for 2 times, and suspending with sterile distilled water of original culture volume to obtain seed for fermentation, wherein the bacterial concentration of the seed liquid is 2x10 9 cfu/mL。
Preparation of a Streptococcus mutans bacterial suspension: dissolving lyophilized powder of Streptococcus mutans CGMCC No.1.2499 (from CGMCC, China) in small amount of sterile distilled water, and taking one ring with inoculating loop to mark on BHI solid culture medium (from OXOID Co., UK)Taking out the bacterial colony after anaerobic culture at 37 ℃ for 24h, picking up a single bacterial colony by using an inoculating loop, putting the single bacterial colony into 10mL BHI liquid culture medium (purchased from OXOID Co., UK), uniformly dispersing the bacterial colony in the liquid culture medium by using a vortex oscillator, taking out the bacterial colony after anaerobic culture at 37 ℃ for 24h, inoculating the bacterial colony into the BHI liquid culture medium by using an inoculation amount of 2% (v/v), centrifuging the culture at 15000rpm for 10min after anaerobic culture at 37 ℃ for 24h, discarding the supernatant, washing the bacterial colony for 2 times by using sterile distilled water, and then re-suspending the bacterial colony to 10 ℃ by using the BHI liquid culture medium 6 cfu/mL.
(b) Preparing soybean milk: weighing dry soybean, adding five times of water, soaking at 37 deg.C for 8 hr, removing water, adding a certain proportion of wet soybean and water into a soybean milk machine to obtain soybean milk with desired solid content, and sterilizing at 121 deg.C for 20min to obtain sterile soybean milk with desired solid content.
(c) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm:
pretreatment of fermented soybean milk: and adjusting the pH value of the fermented soybean milk to 6.80 to obtain a sample to be detected.
Determination of inhibitory Activity: the sterilized magnetic stirrer A (5X 10mM) was immersed in saliva of a healthy population after sterile filtration through a 0.22 μm filter and incubated at 37 ℃ for 4 hours, the magnetic stirrer A was attracted by a sterilized magnetic stirrer B (10X 50mM), rinsed with 10mM PBS (pH7.0), and the magnetic stirrer A was removed and placed in culture wells of a 24-well plate. To the culture well, 1.6mL of BHI broth and 200. mu.L of Streptococcus mutans suspension (10. mu.L) were added 6 cfu/mL) and 400. mu.L of test sample solution or blank control solution (unfermented soymilk), again incubated anaerobically at 37 ℃ for 24h (purchased from Greiner, Germany). And (3) sucking up the magnetic stirrer A by using a magnetic stirrer B, washing by using PBS, taking down the magnetic stirrer A, placing in a culture hole of a 24-hole plate, and naturally drying. Adding 1.6mL of 0.1% (w/v) crystal violet solution into the culture well for dyeing for 15min, sucking up the magnetic stirrer A by using a magnetic stirrer B, leaching by using PBS to remove redundant dyeing solution, taking down the magnetic stirrer A, placing the magnetic stirrer A in the culture well of a 24-pore plate, and naturally drying in the air. Adding 300 μ L anhydrous ethanol into culture well, decolorizing for 15min, transferring 200 μ L into 96-well plate (purchased from Greiner, Germany), and measuring OD600nm with microplate reader to obtain Streptococcus mutansQuantitative data corresponding to the formation amount of the object film. The OD600nm of the blank control and the sample to be tested were measured by the above method, and the inhibition ratio of the biofilm inhibitor was calculated. Biofilm inhibition rate (I):
i ═ OD600nm (blank) -OD600nm (sample to be tested))/OD 600nm (blank) × 100%
2. Preparation of fermented soybean milk
Bacillus bovis (Paenibacillus bovis) CGMCC No.8333 is aseptically inoculated into soybean milk with solid content of 5% (w/w, soybean in water mass percent, the same below) by an inoculation amount of 3% (v/v, seed liquid in fermentation liquid volume percent, the same below), and is subjected to static culture at 37 ℃ for 24h to obtain fermented soybean milk A.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk A is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 47%.
Example 2
1. Materials and methods
(a) Preparation of a suspension of Paenibacillus bovis seed (fermentation strain) and Streptococcus mutans: the same as in example 1.
(b) Preparing soybean milk: weighing dry red bean, adding five times of water, soaking at 37 deg.C for 8 hr, removing water, adding a certain proportion of wet bean and water into a soybean milk machine to obtain soybean milk with desired solid content, and sterilizing at 121 deg.C for 20min to obtain sterile red bean soybean milk with desired solid content.
(c) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
The method comprises the following steps of aseptically inoculating Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 into red bean milk with solid content of 9% (w/w) according to the inoculation amount of 1% (v/v), and carrying out standing culture at 15 ℃ for 48h to obtain fermented soybean milk B.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk B is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 46%.
Example 3
1. Materials and methods
(a) Preparation of a suspension of Paenibacillus bovis seed (fermentation strain) and Streptococcus mutans: the same as in example 1.
(b) Preparing soybean milk: weighing dried mung bean, adding water with five times of mass, soaking at 37 deg.C for 8 hr, removing water, adding wet mung bean and water at a certain ratio into a soybean milk machine to obtain soybean milk with desired solid content, and sterilizing at 121 deg.C for 20min to obtain sterile mung bean soybean milk with desired solid content.
(c) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
The method comprises the following steps of aseptically inoculating Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 into mung bean milk with the solid content of 1% (w/w) according to the inoculation amount of 5% (v/v), and carrying out standing culture at 45 ℃ for 2 hours to obtain fermented soybean milk C.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk C is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 48%.
Example 4
1. Materials and methods
(a) Preparation of paenibacillus bovis seeds (fermentation strain) and streptococcus mutans suspension: the same as in example 1.
(b) Preparing soybean milk: weighing dry broad bean, adding water with the mass of five times, soaking at 37 ℃ for 8 hours, removing water, adding wet bean and water in a certain proportion into a soybean milk machine to prepare soybean milk with the required solid content, and sterilizing at 121 ℃ for 20min to obtain the sterile broad bean soybean milk with the specified solid content.
(c) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
Bacillus bovis (Paenibacillus bovis) CGMCC No.8333 is aseptically inoculated in broad bean milk with solid content of 7% (w/w) by inoculation amount of 2% (v/v), and is kept stand and cultured for 36h at 25 ℃ to obtain fermented soybean milk D.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk D is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 45%.
Example 5
1. Materials and methods
(a) Preparation of paenibacillus bovis seeds (fermentation strain) and streptococcus mutans suspension: the same as in example 1.
(b) Preparing soybean milk: mixing white kidney beans and small white kidney bean dry beans according to a mass ratio of 1: 1, weighing and mixing, adding water with five times the mass, soaking at 37 ℃ for 8 hours, removing the water, adding wet beans and water in a certain proportion into a soybean milk machine to prepare soybean milk with required solid content, and sterilizing at 121 ℃ for 20min to obtain the sterile white kidney bean soybean milk with the specified solid content.
(c) And (3) measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
The method comprises the following steps of aseptically inoculating Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 into white kidney bean milk with solid content of 3% (w/w) according to the inoculation amount of 4% (v/v), and carrying out standing culture at 40 ℃ for 12h to obtain fermented soybean milk E.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk E is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the formation of the streptococcus mutans biofilm is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the formation of the streptococcus mutans biofilm is 49%.
Example 6
1. Materials and methods
(a) Preparation of paenibacillus bovis seeds (fermentation strain) and streptococcus mutans suspension: the same as in example 1.
(b) Preparation of lactobacillus plantarum seeds: dissolving lyophilized powder of Lactobacillus plantarum ATCC14917 with small amount of sterile distilled water, drawing a ring by using an inoculating ring on MRS solid culture medium (purchased from Merck Co. Germany), performing anaerobic culture at 37 ℃ for 24h, taking out a single colony by using the inoculating ring, putting the single colony into 1mL of MRS liquid (purchased from Merck Co. Germany), uniformly dispersing the colony in the liquid culture medium by using a vortex oscillator, performing anaerobic culture at 37 ℃ for 24h, taking out the colony, inoculating the colony into 50mL of MRS liquid in an inoculation amount of 2% (v/v), performing culture at 37 ℃ for 24h, centrifuging the culture at 9,000rpm for 10min, discarding supernatant, washing thallus for 2 times by using sterile distilled water, suspending the thallus by using sterile distilled water with the original culture volume to obtain seed for fermentation, wherein the bacterial concentration of the seed liquid is 1x10 9 cfu/mL。
(c) Preparing soybean milk: the same as in example 1.
(d) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
The fermented soybean milk F is prepared by aseptically inoculating 1.5% (v/v) of the inoculum size of 1.5% (v/v) of the Bacillus bovis (Streptococcus salivarius) CGMCC No.8333 and 1.5% (v/v) of the inoculum size of the Lactobacillus plantarum ATCC14917 into soybean milk with the solid content of 5% (w/w), and performing static culture at 37 ℃ for 24 hours.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk F is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 61%.
Example 7
1. Materials and methods
(a) Preparation of paenibacillus bovis seeds (fermentation strain) and streptococcus mutans suspension: the same as in example 1.
(b) Preparation of lactobacillus plantarum: the same as in example 6.
(c) Preparing soybean milk: the same as in example 1.
(d) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
The fermented soybean milk G is prepared by aseptically inoculating paenibacillus bovis (Streptococcus salivarius) CGMCC No.8333 and Lactobacillus plantarum (Lactobacillus plantarum) ATCC14917 in the inoculation amount of 4.5% (v/v) into soybean milk with the solid content of 3% (w/w) for 12h at 40 ℃.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk G is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is measured, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 58%.
Example 8
1. Materials and methods
(a) Preparation of paenibacillus bovis seeds (fermentation strain) and streptococcus mutans suspension: the same as in example 1.
(b) Preparation of lactobacillus plantarum: the same as in example 6.
(c) Preparing soybean milk: the same as in example 1.
(d) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
The fermented soybean milk H is prepared by aseptically inoculating 0.5% (v/v) of Bacillus bovis (Streptococcus salivarius) CGMCC No.8333 and 4.5% (v/v) of Lactobacillus plantarum ATCC14917 into soybean milk with solid content of 7% (w/w), and performing static culture at 45 ℃ for 2H.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
After the fermented soybean milk H is prepared into a sample to be tested by the method, the inhibitory activity of the sample to the streptococcus mutans biofilm formation is determined, and the result shows that the inhibition rate I of the fermented soybean milk to the streptococcus mutans biofilm formation is 56%.
Effect example 1Product taste and preference test
The fermented soybean milk product A, B, C, D, E, F, G, H prepared in the above examples 1 to 8 was used as a test subject to perform taste test of the product. The number of test persons was 50. Tasting mode: tasting in a mode of unmarked scoring; the color, flavor, taste and nutritional items of the fermented soybean milk product A, B, C, D, E, F, G, H were individually scored, each full score was 25, the average score and the total score thereof were calculated, and the statistical results are recorded in table 1. Meanwhile, according to the opinions given to the overall preference degree of the product, the number of people who like each single product is counted, and the counting result is recorded in the table 2.
TABLE 1 product taste test result data statistics table
TABLE 2 statistical table of product preference test results
As can be seen from the product taste test and the result of the preference degree statistics, the fermented soybean milk with the streptococcus mutans biofilm inhibiting activity prepared by the method in the technical scheme of the invention can be accepted by most consumers in terms of product flavor, mouthfeel and nutrition.
Effect example 2Fermented beans under refrigerated conditionsStability of plasma against Streptococcus mutans biofilm inhibiting Activity
The fermented soybean milk A, B, C, D, E, F, G, H prepared in examples 1 to 8 was stored under refrigeration (4 ℃) for 0, 10, 20 and 30 days and then taken out, and the inhibition rate of each sample against the formation of Streptococcus mutans biofilm was measured, and the results are shown in Table 3.
TABLE 3 stability of inhibitory Effect of fermented Soybean milk on the formation of Streptococcus mutans biofilm under Cold storage conditions
As can be seen from Table 3, after all the fermented soybean milks tested were stored under refrigeration conditions (4 ℃) for 30 days, the inhibition rates of the fermented soybean milks against Streptococcus mutans biofilm were stably maintained at the same level, and the stability was good.
Comparative example 1
The inoculation amount, soybean milk concentration, culture temperature and fermentation time in example 1 were adjusted one by one to obtain fermented soybean milk prepared by the following different methods, and the inhibitory effect of the obtained fermented soybean milk on streptococcus mutans biofilm in each group is shown in table 4.
TABLE 4 inhibitory Effect of fermented soymilk prepared by different methods on the biofilm formation of Streptococcus mutans
From the results shown in table 4, it can be seen that when the inoculation amount, soybean milk concentration, culture temperature and fermentation time in the method for preparing fermented soybean milk were adjusted to be out of the preferred ranges, the fermented soybean milk prepared by Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 could still inhibit the formation of streptococcus mutans biofilm, but the inhibitory effect was significantly reduced.
Therefore, within the preferred range, the inoculation amount, the soybean milk concentration, the culture temperature and the fermentation time are influenced mutually, so that the fermented soybean milk prepared by the Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 has better inhibiting effect on the formation of the streptococcus mutans biofilm.
Comparative example 2
The inoculation amount, soybean milk concentration, culture temperature and fermentation time in example 6 were adjusted one by one to obtain fermented soybean milk prepared by the following different methods, and the inhibitory effect of the obtained fermented soybean milk on streptococcus mutans biofilm in each group is shown in table 5.
TABLE 5 inhibitory Effect of fermented soymilk prepared by different methods on the biofilm formation of Streptococcus mutans
As can be seen from the results shown in table 5, when the inoculation amount, soybean milk concentration, culture temperature and fermentation time in the method for preparing fermented soybean milk were adjusted to be out of the preferred ranges, the fermented soybean milk prepared from Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 and lactobacillus plantarum (l.plantarum) ATCC14917 could still inhibit the formation of streptococcus mutans biofilm, but the inhibitory effect thereof was significantly reduced.
Therefore, within the preferred range, the inoculation amount, soymilk concentration, culture temperature and fermentation time influence each other, so that fermented soymilk prepared from Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 and Lactobacillus plantarum (L.plantarum) ATCC14917 (purchased from ATCC) has a better inhibitory effect on the formation of Streptococcus mutans biofilm.
Comparative example 3
Referring to the method described in example 1, the inhibitory effects of fermented soybean milk prepared from Paenibacillus bovis (Paenibacillus bovis) cgmccno.8333, Lactobacillus plantarum (Lactobacillus plantarum) ATCC14917, streptococcus thermophilus (s.thermophilus) ST-BODY-3 (provided by the company hansen, koch.), Lactobacillus casei (l.casei) ATCC 393 (purchased from ATCC) on the biofilm formation of streptococcus mutans were compared as follows:
1. materials and methods
(a) Preparation of seeds (fermentation strain):
preparation of Paenibacillus bovis seeds (fermentation strain): the same as in example 1.
Preparation of lactobacillus plantarum seeds (zymocyte species): the same as in example 6.
Preparation of lactobacillus casei seeds: dissolving freeze-dried powder of lactobacillus casei ATCC 393 in a small amount of sterile distilled water, taking a ring by using an inoculating ring, scribing on an MRS solid culture medium (purchased from Merck Co, Germany), carrying out anaerobic culture at 37 ℃ for 24h, taking a single colony by using the inoculating ring, putting the single colony into 1mL of MRS liquid (purchased from Merck Co, Germany), uniformly dispersing the colony in the liquid culture medium by using a vortex oscillator, carrying out anaerobic culture at 37 ℃ for 24h, inoculating the single colony into 50mL of MRS liquid by using an inoculation amount of 2% (v/v), carrying out culture at 37 ℃ for 24h, centrifuging a culture at 9,000rpm for 10min, discarding a supernatant, washing thalli for 2 times by using the sterile distilled water, and suspending by using the sterile distilled water with the original culture volume to obtain seeds for fermentation.
Preparation of streptococcus thermophilus seeds: dissolving the freeze-dried powder of the streptococcus thermophilus ST-BODY-3 with a small amount of sterile distilled water, drawing a ring by using an inoculating ring, streaking the ring on an M17 solid culture medium (purchased from Merck Co. Germany), carrying out anaerobic culture at 40 ℃ for 24h, taking out, picking a single colony by using the inoculating ring, putting the single colony into 1mL of M17 liquid (purchased from Merck Co. Germany), uniformly dispersing the colony in the liquid culture medium by using a vortex oscillator, carrying out anaerobic culture at 40 ℃ for 24h, taking out, inoculating the single colony into 50mL of M17 liquid by using an inoculation amount of 2% (v/v), carrying out culture at 40 ℃ for 24h, centrifuging the culture at 9,000rpm for 10min, discarding the supernatant, washing the thalli for 2 times by using sterile distilled water, and suspending the thalli by using the sterile distilled water with the original culture volume to obtain seeds for fermentation.
Preparation of a Streptococcus mutans bacterial suspension: the same as in example 1.
(b) Preparing soybean milk: the same as in example 1.
(c) Measuring the inhibition rate of the fermented soybean milk on the formation of the streptococcus mutans biofilm: the same as in example 1.
2. Preparation of fermented soybean milk
Inoculating each strain into soybean milk with solid content of 5% (w/w) at 3% (v/v) aseptically, and culturing (anaerobic culture of Paenibacillus bovis, Lactobacillus plantarum and Lactobacillus casei at 37 deg.C, and anaerobic culture of Streptococcus thermophilus at 40 deg.C) for 24h to obtain corresponding fermented soybean milk.
3. Determination of inhibition rate of fermented soybean milk on formation of streptococcus mutans biofilm
The inhibitory effect of fermented soymilk prepared from different strains on the formation of Streptococcus mutans biofilm is shown in Table 6:
TABLE 6 inhibitory Effect of fermented soymilk prepared with different strains on the biofilm formation of Streptococcus mutans
As can be seen from Table 6, the fermented soybean milk prepared from other strains did not have the inhibitory activity against Streptococcus mutans biofilm, whereas the inhibitory activity against Streptococcus mutans biofilm of the fermented soybean milk prepared from Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 was very significant.
As can be seen from example 6, the inhibitory activity of the fermented soybean milk prepared by combining Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 with that of the fermented soybean milk prepared by combining lactobacillus plantarum (l.plantatarum) ATCC14917 (purchased from ATCC) was 61%, and the inhibitory activity of the fermented soybean milk prepared by combining Paenibacillus bovis (Paenibacillus bovis) CGMCC No.8333 with that of the fermented soybean milk prepared by combining bacillus bovis (Paenibacillus bovis) CGMCC No.8333 with that of the fermented soybean milk prepared by combining lactobacillus plantarum (l.plantatarum) ATCC14917 (purchased from ATCC) was significant against the streptococcus mutans biofilm.
Comparative example 4
1. Materials and methods
(a) The preparation of the paenibacillus bovis CGMCC No.8333 and the streptococcus mutans bacterial suspension comprises the following steps: the same as in example 1. Preparation of lactobacillus plantarum ATCC 14917: the same as in example 6.
(b) And (3) determining the inhibition rate of the fermentation liquor on the formation of the streptococcus mutans biofilm:
pretreatment of fermentation liquor: and adjusting the pH value of the fermentation liquor to 6.80 to obtain a sample to be detected.
Determination of inhibitory Activity: the assay was performed with unfermented medium as a blank, according to the method described in example 1.
2. Preparation of Paenibacillus bovis single-bacterium fermentation broth and mixed fermentation broth of Paenibacillus bovis and Lactobacillus plantarum
Aseptically inoculating paenibacillus bovis CGMCC No.8333 in an inoculum size of 3% (v/v) into an M17 liquid culture medium containing 1% of sucrose, and anaerobically culturing at 37 ℃ for 24h to obtain a fermentation liquid;
the method comprises the following steps of carrying out aseptic inoculation on a M17 liquid culture medium containing 1% of sucrose by using 1.5% (v/v) inoculation amount of paenibacillus bovis CGMCC No.8333 and 1.5% (v/v) inoculation amount of Lactobacillus plantarum ATCC14917, and carrying out anaerobic culture at 37 ℃ for 24 hours to obtain a fermentation liquid.
3. Determination of inhibition rate of fermentation liquor on formation of streptococcus mutans biofilm
The inhibitory activity of the fermentation liquor on the formation of the streptococcus mutans biofilm is measured after the fermentation liquor is prepared into a sample to be detected by the method, and the result shows that the inhibitory rates of the single-bacterium fermentation liquor of the paenibacillus bovis, the mixed fermentation liquor of the paenibacillus bovis and the lactobacillus plantarum on the formation of the streptococcus mutans biofilm are respectively 9% and 11%.
Comparative example 5
The pH of the fermented soybean milk A, B, C, D, E, F, G, H prepared in examples 1 to 8 was adjusted to 6.80, and then centrifuged at 10,000rpm at 4 ℃ for 10min to obtain a supernatant fraction as a sterilized fermented soybean milk (sample to be tested). The inhibition rate of biofilm formation by Streptococcus mutans was determined for the above samples according to the method described in example 1, and the results are shown in Table 7.
TABLE 7 inhibitory Effect of aseptically fermented Soybean milk on Streptococcus mutans biofilm formation
As can be seen from Table 7, the aseptic fermented soybean milk still has strong inhibitory activity on the biofilm formation inhibition rate of Streptococcus mutans, which indicates that the inhibitory effect is derived from the products of the strains after metabolizing the soybean milk. However, the biofilm inhibition effect of each group of the aseptic fermented soybean milk is lower than that of the sample group before the aseptic treatment, which shows that the biofilm inhibition effect of the fermented soybean milk before the aseptic treatment comes from the combined action of the viable bacteria and the metabolite.
The preparation method of the fermented soybean milk, the prepared fermented soybean milk and the application thereof provided by the invention are described in detail above. The principles and embodiments of the present invention are explained herein using specific examples, which are presented only to assist in understanding the method and its core concepts. It should be noted that, for those skilled in the art, it is possible to make various improvements and modifications to the present invention without departing from the principle of the present invention, and those improvements and modifications also fall within the scope of the claims of the present invention.
Claims (5)
1. A method for preparing fermented soybean milk by using Paenibacillus bovis is characterized by comprising the following steps: inoculating paenibacillus bovis CGMCC No.8333 serving as a zymophyte agent into the soybean milk for fermentation to obtain fermented soybean milk; the fermentation temperature is 15-45 ℃, and the fermentation time is 2-48 h;
the fermentation inoculum also comprises lactobacillus plantarum ATCC14917, and the inoculation ratio of the lactobacillus plantarum ATCC14917 to paenibacillus bovis CGMCC No.8333 is 1-9: 1-9;
the mass percentage content of the solid in the soybean milk is 1-9%;
the inhibition rate of the fermented soybean milk on the formation of streptococcus mutans biofilm is more than or equal to 46%.
2. The method for preparing fermented soybean milk according to claim 1, wherein the inoculation amount of the fermentation inoculant is 2x10 7 ～1x10 8 cfu/mL。
3. The method for preparing fermented soybean milk according to claim 1, wherein the soybean milk is at least one of soybean, red bean, mung bean, broad bean, white kidney bean or small white kidney bean milk.
4. The method for preparing fermented soybean milk according to claim 1, wherein the fermentation is static culture.
5. A fermented soybean milk prepared from Paenibacillus bovis, characterized by being prepared by the preparation method of any one of claims 1 to 4, wherein the inhibition rate of the fermented soybean milk on the formation of streptococcus mutans biofilm is not less than 46%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910435092.9A CN110122577B (en) | 2019-05-23 | 2019-05-23 | Method for preparing fermented soybean milk from paenibacillus bovis, prepared fermented soybean milk and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910435092.9A CN110122577B (en) | 2019-05-23 | 2019-05-23 | Method for preparing fermented soybean milk from paenibacillus bovis, prepared fermented soybean milk and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110122577A CN110122577A (en) | 2019-08-16 |
| CN110122577B true CN110122577B (en) | 2022-09-02 |
Family
ID=67572793
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910435092.9A Active CN110122577B (en) | 2019-05-23 | 2019-05-23 | Method for preparing fermented soybean milk from paenibacillus bovis, prepared fermented soybean milk and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110122577B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106509105A (en) * | 2016-11-07 | 2017-03-22 | 光明乳业股份有限公司 | Fermented milk, preparation method thereof and fermented milk powder |
| CN108728497A (en) * | 2017-04-24 | 2018-11-02 | 光明乳业股份有限公司 | A kind of streptococcus mutans inhibitor and preparation method thereof |
-
2019
- 2019-05-23 CN CN201910435092.9A patent/CN110122577B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106509105A (en) * | 2016-11-07 | 2017-03-22 | 光明乳业股份有限公司 | Fermented milk, preparation method thereof and fermented milk powder |
| CN108728497A (en) * | 2017-04-24 | 2018-11-02 | 光明乳业股份有限公司 | A kind of streptococcus mutans inhibitor and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| Probiotic Lactobacillus sp. inhibit growth, biofilm formation and gene expression of caries-inducing Streptococcus mutans;Reham Wasfi，等;《J. Cell. Mol. Med.》;20181231;第22卷(第3期);第1972-1983页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110122577A (en) | 2019-08-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110100899B (en) | Method for preparing fermented soybean milk powder from lactobacillus fermentum, prepared fermented soybean milk powder and application | |
| EP2217083B1 (en) | Beverage compositions | |
| JP4772131B2 (en) | Method for producing fermented milk using novel lactic acid bacteria | |
| CN110122576B (en) | Method for preparing fermented soybean milk by lactobacillus fermentum, prepared fermented soybean milk and application | |
| CH649196A5 (en) | PROCESS FOR THE MANUFACTURE OF YOGURT. | |
| KR100471631B1 (en) | Bifidobacterium breve and fermented soymilk prepared with the same | |
| FI76373B (en) | FOERFARANDE FOER FRAMSTAELLNING AV EN ODLING SOM INNEHAOLLER LEVANDE CELLMASSA AV BIFIDOBAKTERIER OCH MJOELKSYRABAKTERIER. | |
| CN112126599B (en) | High-density culture method of lactobacillus helveticus, preparation of high-activity bacterium powder and application of high-density culture method | |
| CN106509105A (en) | Fermented milk, preparation method thereof and fermented milk powder | |
| CN110122581B (en) | Method for preparing fermented soybean milk from leuconostoc mesenteroides, prepared fermented soybean milk and application | |
| CN110012941B (en) | Method for preparing fermented soybean milk by using paenibacillus, fermented soybean milk prepared by method and application of fermented soybean milk | |
| US4210672A (en) | Preparation of yogurt | |
| CN110122578B (en) | Method for preparing fermented soybean milk powder from leuconostoc mesenteroides, prepared fermented soybean milk powder and application | |
| CN107441121B (en) | Fermentation crude extract of streptococcus salivarius, and preparation method and application thereof | |
| CN110122577B (en) | Method for preparing fermented soybean milk from paenibacillus bovis, prepared fermented soybean milk and application | |
| Sanni et al. | Microbiological evaluation of Ghanaian maize dough co-fermented with cowpea | |
| CN107410479B (en) | Fermented milk and preparation method thereof | |
| CN110150387B (en) | Method for preparing fermented soybean milk powder from paenibacillus, fermented soybean milk powder and application | |
| CN108813045A (en) | A kind of preparation method of pectase honey lemon tea | |
| CN110951654A (en) | Lactobacillus plantarum proliferation agent and application thereof | |
| CN101886043B (en) | Composite yoghurt starter and application thereof | |
| CN110122579A (en) | It is a kind of the method for fermented soybean milk to be prepared by streptococcus salivarius and the fermented soybean milk prepared and application | |
| CN110122580A (en) | A kind of fermented soybean milk powder for being prepared the method for fermented soybean milk powder by streptococcus salivarius and being prepared and application | |
| CN110122575A (en) | A kind of fermented soybean milk powder for being prepared the method for fermented soybean milk powder by ox series bacillus and being prepared and application | |
| CN108004176A (en) | A kind of application of Lactobacillus rhamnosus LRH09, leavening prepared therefrom and leavening |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |