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CN-116200450-B - Screening method and system for ecological coexistence probiotics

CN116200450BCN 116200450 BCN116200450 BCN 116200450BCN-116200450-B

Abstract

The invention discloses a screening method and a screening system of ecological coexistence probiotics, wherein the screening method comprises the steps of obtaining a single strain with a preset function and high flux based on high-flux screening of multiple targets, combining the strains according to a function combination mode to obtain the probiotics combination, performing in-vitro co-culture, detecting an OD value of the probiotics combination, and judging that the probiotics combination has high possibility of in-vivo collaborative symbiosis according to the fact that if the OD value of the probiotics combination is larger than a preset threshold value, otherwise, the possibility of in-vivo collaborative symbiosis small, and the preset threshold value is 1.9-2.1. The screening method carries out probiotic combination in a functional combination mode, screens by adopting a threshold method, screens according to the principle that the in-vivo collaborative symbiosis possibility is high when the OD value of the probiotic combination is larger than a preset threshold value, and compared with random combination, the screening method and the screening system can rapidly screen the probiotic combination.

Inventors

  • LIU ZHI
  • DENG YUN

Assignees

  • 华中科技大学

Dates

Publication Date
20260508
Application Date
20230106

Claims (6)

  1. 1. The screening method of the ecological coexistence probiotics is characterized by comprising the following steps of: based on high-throughput screening of multiple targets, the probiotic combination with weight loss screening is obtained, and according to the single strain with high throughput of pancreatic lipase inhibition, bile salt hydrolase activity, GLP-1 hormone secretion stimulation and preadipocyte differentiation inhibition functions; Combining different high-flux strains according to a functional combination mode to obtain a probiotic combination, wherein the high-flux strains are selected from bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, bifidobacterium animalis, bifidobacterium bifidum, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus salivarius and/or lactococcus lactis; Screening probiotic combinations with weight loss, wherein the probiotic combinations are performed according to four functions of pancrelipase inhibition, bile salt hydrolase activity, GLP-1 hormone secretion stimulation and preadipocyte differentiation inhibition, and the number of the combinations is not more than four; in vitro co-culture is carried out, OD value of probiotic combination is detected, in vitro experiment is used for evaluating in vivo application, and judgment and screening are carried out according to the following principle: If the OD value of the probiotic combination is larger than a preset threshold, judging that the probability of the synergistic symbiosis of the probiotic combination in the body is high, otherwise, the probability of the synergistic symbiosis in the body is low, wherein the preset threshold is 1.9-2.1; The OD value is specifically measured as follows: the same probiotic combination was cultivated to logarithmic growth phase under anaerobic conditions and the OD value at 600nm was measured.
  2. 2. The method for screening of ecologically coexisting probiotics according to claim 1, characterized in that the OD value is detected specifically as follows: Inoculating the same probiotic combination according to the inoculum size of a preset proportion, culturing to a logarithmic growth phase, and detecting an OD value under the condition of corresponding wavelength, wherein the preset proportion is 1% -3%.
  3. 3. The screening system of the ecological coexistence probiotics is characterized by comprising an OD value acquisition module and a judgment module; The OD value acquisition module is used for acquiring OD values of probiotic combinations and submitting the OD values to the judgment module, wherein the probiotic combinations are carried out according to four functions of pancrelipase inhibition, bile salt hydrolase activity, GLP-1 hormone secretion stimulation and preadipocyte differentiation inhibition, and the number of the combinations is not more than four, and the probiotic is selected from bifidobacterium adolescentis, bifidobacterium longum, bifidobacterium breve, bifidobacterium animalis, bifidobacterium bifidum, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus salivarius and/or lactococcus lactis; The OD value is specifically measured as follows: Culturing the same probiotic combination to logarithmic growth phase under anaerobic condition, and detecting OD value under 600 nm; the judging module is used for replacing in-vivo application evaluation through in-vitro experiments according to the obtained OD value, and specifically judges according to the following principle: If the OD value of the probiotic combination is larger than a preset threshold, judging that the probability of the synergistic symbiosis of the probiotic combination in the body is high, otherwise, the probability of the synergistic symbiosis in the body is low, and the preset threshold is 1.9-2.1.
  4. 4. The system for screening for co-existing probiotics according to claim 3, wherein the OD value obtaining module obtains the obtained OD value according to the following detection: Inoculating the same probiotic combination according to the inoculum size of a preset proportion, culturing to a logarithmic growth phase, and detecting an OD value under the condition of corresponding wavelength, wherein the preset proportion is 1% -3%.
  5. 5. The system for screening for co-existing probiotic bacteria according to claim 4, wherein said OD value obtaining module, when screening for probiotic bacteria combinations with weight loss, obtains said OD value according to the following detection: the same probiotic combination was inoculated at an inoculum size of 1% and cultured under anaerobic conditions to logarithmic growth phase.
  6. 6. The system for screening for co-existing probiotics according to claim 5, wherein the judgment module judges that the probability of co-symbiosis of the probiotic combination in vivo is high if the OD value of the probiotic combination is greater than 1.9, and judges that the probability of co-symbiosis in vivo is low if the OD value of the probiotic combination is less than 1.9.

Description

Screening method and system for ecological coexistence probiotics Technical Field The invention belongs to the technical field of biology, and particularly relates to a screening method and a screening system of ecological coexistence probiotics. Background Probiotics, when ingested in sufficient amounts, have beneficial health effects on the host. However, single strains often have general effects, and researches show that the effects of combining probiotics are generally better than those of single strains, but the effects of the probiotics combined are uncertain. The existing probiotics combination generally combines 2-3 bacteria, the screening way is single, the strains obtained by screening are generally only verified on single bacterial effect, or the strains are simply and randomly combined, and the combination of other strains has no reference significance. Moreover, this random combination of strains does not always have a more beneficial effect, as found in studies of probiotic improvement of obesity, although the administration of both of the two probiotics alone reduced the weight of the fed mice, the combination did not have a significant anti-obesity effect, and even the IMV B-7280 strain alone was found to give a better effect than the VKL/VKB/IMV B-7280 combination. This is mainly because there may be various interactions between different strains, such as metabolic dependency, nutritional competition, bacteriocin antagonism, etc., so that the strains do not coexist well in vivo, and even the interactions between the strains may counteract or even reduce the beneficial function of the probiotics. If the function verification and in-vivo experiment verification are carried out on the probiotic combination, the problem of functional uncertainty caused by random combination of probiotics can be solved to a certain extent, but the verification period is long, time and labor are wasted, and especially the probiotic combination in the aspect of new efficacy has extremely large workload and long time. Therefore, how to quickly and effectively screen out the probiotic combination which can coexist ecologically and has obvious effect is a difficult problem to be solved. Disclosure of Invention Aiming at the defects or improvement demands of the prior art, the invention provides a screening method and a screening system of ecological coexistence probiotics, which aim to screen out the probiotic combination with high symbiotic possibility in vivo by combining strains according to functional combinations and using the OD value of the probiotic combination to be larger than a preset threshold value, wherein the threshold value is 1.9-2.1, so that the technical problems of complex operation and long period of the existing screening of the probiotic combination with good symbiotic property in vivo are solved. To achieve the above object, according to one aspect of the present invention, there is provided a screening method of ecologically coexisting probiotics, comprising the steps of: high-flux screening based on multiple targets to obtain a single strain with preset function and high flux; Combining different high-flux strains according to a functional combination mode to obtain a probiotic combination; in-vitro co-culture is carried out, OD values of probiotic combinations are detected, and judgment and screening are carried out according to the following principle: If the OD value of the probiotic combination is larger than a preset threshold, judging that the probability of the synergistic symbiosis of the probiotic combination in the body is high, otherwise, the probability of the synergistic symbiosis in the body is low, and the preset threshold is 1.9-2.1. Preferably, the OD value of the screening method of the ecological coexistence probiotics is specifically detected as follows: Inoculating the same probiotic combination according to the inoculum size of a preset proportion, culturing to a logarithmic growth phase, and detecting an OD value under the condition of corresponding wavelength, wherein the preset proportion is 1% -3%. Preferably, the method for screening the ecological coexistence probiotic comprises performing in vitro co-culture of a probiotic combination having two or more functions of pancreatic lipase inhibition, biliary hydrolase activity, GLP-1 hormone secretion stimulation and preadipocyte differentiation inhibition, and detecting an OD value when screening the probiotic combination having weight loss. Preferably, the method for screening the probiotic bacteria coexisting ecologically comprises performing a probiotic bacteria combination according to four functions of pancreatic lipase inhibition, bile salt hydrolase activity, GLP-1 hormone secretion stimulation and preadipocyte differentiation inhibition, and detecting OD value at a wavelength of 600 nm. Preferably, the method for screening the probiotic bacteria coexisting ecologically, when screening the probiotic bacteria c