CN114568699B - Composition containing galactooligosaccharides and probiotics and application thereof - Google Patents

Composition containing galactooligosaccharides and probiotics and application thereof Download PDF

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CN114568699B
CN114568699B CN202011376993.4A CN202011376993A CN114568699B CN 114568699 B CN114568699 B CN 114568699B CN 202011376993 A CN202011376993 A CN 202011376993A CN 114568699 B CN114568699 B CN 114568699B
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bifidobacterium
infancy
composition
bifidobacterium longum
subspecies
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CN114568699A (en
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刘彪
石羽杰
洪维鍊
刘伟贤
赵雯
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Inner Mongolia Yili Industrial Group Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/127—Fermented milk preparations; Treatment using microorganisms or enzymes using microorganisms of the genus lactobacteriaceae and other microorganisms or enzymes, e.g. kefir, koumiss
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/16—Agglomerating or granulating milk powder; Making instant milk powder; Products obtained thereby
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/385—Concentrates of non-alcoholic beverages
    • A23L2/39—Dry compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • 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
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • 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
    • A23V2400/51—Bifidobacterium
    • A23V2400/515—Animalis
    • 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
    • A23V2400/51—Bifidobacterium
    • A23V2400/533—Longum
    • 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
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides a composition containing galacto-oligosaccharide and probiotics and application thereof. The probiotics in the composition containing the galactooligosaccharide and the probiotics are bifidobacterium animalis BB12 and bifidobacterium longum subspecies infantis GB-1496. The invention also provides application of the composition in promoting proliferation of bifidobacterium longum subspecies infantis GB-1496. According to the invention, the research discovers that the compounded bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 can obviously promote the proliferation of bifidobacterium infancy, and the number of the proliferation is obviously higher than that of single combination of the GOS and the bifidobacterium animalis BB12 or the bifidobacterium longum subspecies infancy GB-1496. Moreover, in case a combination of GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 is used, bifidobacterium subspecies infancy GB-1496 exhibits advantageous proliferation.

Description

Composition containing galactooligosaccharides and probiotics and application thereof
Technical Field
The invention relates to a composition containing galacto-oligosaccharides and probiotics and application thereof, belonging to the technical field of health-care food.
Background
The intestinal tract system of human body contains a large amount of bacteria, the total amount of which is more than 100 times of that of human body cells, and the bacteria form intestinal microecology of human body. More and more researches indicate that intestinal microecology is indistinguishable from human disease health, and especially early in life, intestinal microecology has an important influence on life health [1].
Previous studies have shown that different feeding patterns have an important effect on early intestinal micro-ecology in infants. The ratio of bifidobacteria to lactobacilli in the faeces of infants in the formula fed group was significantly lower than in infants in the breast fed group. By adding probiotics and prebiotics to infant formula, the intestinal microecology of the infant can be regulated, approaching the intestinal microecology profile of breast-fed infants, especially the proportion of bifidobacteria [2].
BB12 belongs to bifidobacterium animalis and is a probiotic with the most extensive research history and product application. BB-12 is resistant to gastric acid and bile and can survive in an environment of pH2, mainly because BB12 has a gene encoding bile salt hydrolase that hydrolyzes bound bile salts in the intestinal tract to form amino acids and free bile acids [4]. There have been several studies demonstrating that BB-12 can survive the gastrointestinal tract and that BB-12 can be detected in the feces of term infants fed with the partially hydrolyzed whey formula supplemented with BB-12 at 6 weeks. BB-12 is detected in the feces of a subject after a healthy adult consumes BB-12 or BB-12+galacto-oligosaccharides for one week. The BB-12 level in the feces of the yogurt containing BB-12 and inulin was significantly higher than that of the control group, and 90% of BB-12 was viable bacteria [7] for adults for 4 weeks. Clinical trials in different populations have shown that BB12 has good safety and tolerability, including children 1-5 years [8], healthy adults with recent antibiotics [9], women from early gestation to lactation [10].
There are a number of clinical studies reporting various benefits that it brings to the human body. First, BB12 may bring about intestinal comfort. The infants with intestinal colic are supplemented with BB12 (10≡9 cfu/day), and crying time is reduced by more than 50% and more than that of the control group [11]. BB-12 was added to low lactose and whey protein partially hydrolyzed formula to feed infants with hernia for 1 month, crying of the infants in the intervening group continued significantly lower than the control group, and all parameters of the quality of life (QOL) questionnaire improved significantly [12]. For elderly and healthy adults in nursing homes, supplementation with BB-12 increases intestinal peristalsis [37] and frequency of bowel movement, especially the uncomfortable gastrointestinal symptoms that are insufficient to go to hospital consultation [38]. Second, BB12 is beneficial for immune health. The milk powder added with BB-12 has a preventive effect on acute diarrhea when fed to infants [39], and domestic clinical experiments show that compared with the treatment effects of BB-12, bacillus subtilis bigeminal viable bacteria (mammy love) and a blank control group in patients with children diarrhea, the results show that BB-12 is effective in treatment, and the effect is not significantly different from mammy love [40]. Partially hydrolyzed whey protein-supplemented milk powder supplemented with BB12 (10≡6cfu) was studied in healthy term infants, resulting in an increase in fecal IgA total in vaginally delivered formula fed infants. In formula fed infants produced by caesarean section, anti-rotavirus specific IgA was also elevated, and anti-poliovirus specific IgA was elevated in the feces of all probiotic-supplemented infants [41]. The premature infants were fed milk powder containing BB12 (1.6X10A 9 on the first three days, 4.8X10A 9cfu on the fourth day), and the pH of the faeces was significantly lower than that of the placebo group. Short chain fatty acids in the infant's faeces were elevated, with lower levels of calprotectin, whereas the faecal IgA levels were higher in this group compared to placebo [43]. BB-12 maintains the health of the gastrointestinal flora of premature infants, and premature infants were intervened with BB-12 for 3 weeks, both by fluorescence in situ hybridization and bacterial culture, and were found to significantly increase the number of intestinal bifidobacteria, while reducing the numbers of enterobacteria and clostridia [44]. Following long-term follow-up of neonates who were post-natally supplemented with BB-12, the respiratory tract infection was found to be significantly lower in infants at 8 months than in the control group [45], while the respiratory tract infection was still significantly lower in infants at 2 years [46]. The yogurt ice cream added with the milk BB12 before the fermentation process can improve IL-2 secretion and NK cytotoxicity and has less upper respiratory tract infection fever days [47] when the yogurt ice cream is eaten by healthy adults. Thirdly, BB requests beneficial oral health. BB12 tablets (10≡10 cfu/day) were administered to infants from 1-2 months to 2 years old, and the oral mutans streptococcus number was evaluated at 8 months, with BB12 group being the lowest [48].
Breast milk contains not only abundant nutrients but also viable microbial flora. For breast-fed infants, breast milk micro-ecology is a key influencing factor for the intestinal micro-ecology development of infants. The most abundant species in the intestinal microecology of breast-fed infants is bifidobacteria in early life. Further microbiological analysis revealed that the intestinal flora of breast-fed infants is different from that of adults and is enriched with bifidobacterium infantis. Previous studies have shown that the efficacy of bifidobacterium infantis mainly includes the following aspects:
first, bifidobacterium infantis has a strong ability to utilize HMO, has advantages in intestinal colonisation, and unlike other bifidobacteria infantis contains a broader range of genes related to HMO transport and metabolism in the genome [13-19], and in vitro studies bifidobacterium infantis grows better in the presence of HMO than other strains [19,25], so bifidobacterium infantis strains have unique advantages over other conventional probiotics. The bifidobacterium infantis can also metabolize the produced short chain fatty acids to reduce intestinal pathogens and promote the growth and development of intestinal epidermal cells [36]. Short chain fatty acids are commonly acetic, propionic, butyric and lactic acids, which, besides being absorbed as energy substances by intestinal cells, contribute to the integrity of the intestinal epidermal cells [42].
Second, bifidobacterium infantis have anti-inflammatory effects, and intestinal inflammation in premature infants is an important cause of reduced Necrotizing Enterocolitis (NEC) in premature infants. The study using intestinal cells as a model shows that the bifidobacterium infantis can induce the intestinal cells to express anti-inflammatory cytokines [20-23], and the rat animal experiment shows that the addition of the bifidobacterium infantis can reduce the inflammation of NEC [24]. Bifidobacteria are often absent in the gut of premature infants [26], and the incidence of NEC in breast-fed premature infants is significantly lower than in formula-fed infants [27]. Unlike term breast milk, preterm breast milk contains a higher concentration of fucosylated HMO [28]. Comparing the feeding effect of bifidobacterium infantis and bifidobacterium lactis in the premature infant population, it was found that the ability of bifidobacterium infantis to colonize was stronger than that of bifidobacterium lactis, both in formula feeding and breast feeding, whereas the highest number of bifidobacterium faecalis was that of the group of bifidobacterium infantis fed breast milk [29]. A series of clinical studies have found that supplementation of preterm infants with bifidobacteria infancy can reduce the risk of NEC occurrence [31-35].
Finally, the advantage of adding bifidobacterium infantis is also reflected in immunity, and in a full term clinical study of Bengala, it is found that the number of bifidobacterium infantis in the intestinal tract of the infant is positively correlated with the vaccine response in the early four months of life, that is, the higher the number of bifidobacterium infantis is, the higher the corresponding index of the vaccine response is [30-31], and the trend still exists in the 2 years after birth [31], and it is speculated that bifidobacterium infantis helps to activate immune function and immune memory function.
Prebiotics are a class of carbohydrates that are indigestible by the human body and which interact with the intestinal flora in the intestinal tract of the host to produce substances or effects beneficial to the host [3]. More prebiotics currently studied include: galacto-oligosaccharides, fructo-oligosaccharides, inulin, isomerized lactose, polydextrose, xylo-oligosaccharides, and the like.
The structure of Galactooligosaccharides (GOS) is such that 1-7 galactosyl groups are linked to the galactosyl side of lactose mainly by beta-1, 4-glycosidic linkages [49]. The disaccharide lactose is used as a reaction substrate, is converted by transglycosylation of beta-D-galactosidase, and is hetero-oligosaccharide formed by combining galactose and glucose. The glucose group and the galactose group in the GOS structure are connected by beta-1, 4-glycosidic bond, and adjacent galactose groups are connected by beta-1, 3, beta-1, 4 and beta-1, 6-glycosidic bond, wherein the beta-1, 4-glycosidic bond is mainly connected, and the oligosaccharide is mainly galacto-triose, tetraose, a small amount of pentasaccharide, hexasaccharide and the like. In addition, some microbial beta-D-galactosidases catalyze the hydrolysis of glucose and galactose, which are produced from the substrate lactose, to synthesize galactooligosaccharides with beta-1, 3 and beta-1, 6-glycosidic linkages that differ in structure from lactose. The animal milk contains a small amount of GOS, and the human breast milk has a slightly higher content of GOS [50], so that the milk has important physiological effects on the establishment of microbial flora in the intestinal tracts of animals and infants in the lactation period.
The prebiotic effect of GOS has been demonstrated in a number of clinical studies. First, as a prebiotic. The infant formula of GOS stimulates the growth of bifidobacteria and lactobacilli in the intestines and has faecal properties similar to those of breast-fed term infants [51-52]. The bifidobacteria in feces can be increased by supplementing healthy people over 50 years old with 5g GOS per day [53]. In another healthy adult study, the human fecal bifidobacteria levels were higher in GOS than in placebo-controlled groups, especially after amoxicillin administration [54]. The combined use of GOS with a novel iron agent (half of the common iron agent at the iron content) can improve the intestinal flora, especially the bifidobacterium and lactobacillus quantity, and the effect of preventing anemia is not significantly different from that of the common iron agent [55]. Administration of 15g of GOS daily to obese pre-diabetic patients increases the bifidobacteria population in the gut but does not affect insulin sensitivity and associated energy metabolism [56]. By regulating intestinal microecology, GOS can improve defecation. Elderly women (69-87 years old) with constipation problems can relieve constipation by taking 9g GOS daily, and increase the number of times of defecation per week [57]. The addition of low levels of GOS (2.4 g/l) to infant formulas increases the frequency of defecation, decreases the fecal pH, and stimulates bifidobacteria and lactobacilli in the gut to levels found in breast-fed infants [58]. Another clinical study of infants found that increasing GOS (4-8 g/l) content improved stool consistency in infants [60]. The addition of GOS (5 g/L) to the two-stage formula has a positive effect on the bifidobacterium flora and stool consistency of the infant during the weaning supplementation period [59]. Second, GOS is able to stimulate boost immunity. Supplementing elderly women (65-85 years old) with lactoferrin (1 g/day), GOS (2.64 g/day) and vitamin D (20 ug vitamin D3/day) in combination improves immunomodulation and reduces the risk of infection [61]. Healthy infants were fed prebiotic milk powder (scGOS: pdx=1:1), and at 6 months, 8 months, 10 months, 12 months, the weight of the control formula was higher than that of the probiotic milk powder and breast milk group. The duration of diarrhea, the number of constipation and the number of respiratory infections within 1 year old all differed significantly, with the control formula > probiotic formula > breast milk group. Healthy infants (at least one parent had allergic problems) were recruited and the prebiotic milk powder (4 g/L, scGOS: pdx=1:1) was fed to at least one respiratory tract infection and the average number of respiratory tract infections occurred by 48 weeks, with the prebiotic milk powder group being lower than the control formula group [62]. Fatigue is a predictor of cardiovascular events in End Stage Renal Disease (ESRD) patients receiving hemodialysis treatment. The administration of 50ml of a liquid containing 10 mg of vitamin B1, 1.8 mg of vitamin B2, 15 mg of niacin, 10 mg of vitamin B6, 30 mg of vitamin B12, 0.5 mg of folic acid, 60 mg of vitamin C, 500 mg of carnitine, 30 mg of coenzyme Q10 (coenzyme Q10), 5g of natural galactooligosaccharide and 8 mg of zinc per day can regulate the immune and autonomic nervous dysfunction of hemodialysis patients [63]. Fourth, at the age of 10-13 years old girls, eating 5g GOS per day increases calcium absorption, possibly mediated by intestinal microbiota, especially bifidobacteria [64]. Fifth, supplementing healthy adults with 1g of gos twice daily can improve skin conditions, including skin dehydration and wrinkling [65].
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Disclosure of Invention
In order to solve the technical problems, the invention aims to provide a composition capable of promoting proliferation of bifidobacterium longum subspecies infancy GB-1496 and application thereof.
To achieve the above object, the present invention provides a composition comprising galactooligosaccharides and probiotics, wherein the probiotics are bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496.
According to a specific embodiment of the present invention, in the composition of the present invention, the bifidobacterium longum subspecies infancy GB-1496 is bifidobacterium subspecies infancy (Bifidobacterium longum subsp. The strain is preserved in China Center for Type Culture Collection (CCTCC), and the preservation unit address is: 430072, university of martial arts, chinese; preservation date: 04 th 2011; preservation number: cctccc NO: m2011122, classification nomenclature: bifidobacterium longum subspecies infantis (Bifidobacterium longum subsp.infantis).
The bifidobacterium longum subspecies infancy CCTCC No. M201122 has better free radical scavenging capability and reducing capability, and can induce Caco-2 cells to increase the expression of antioxidant enzyme, so that the bifidobacterium longum subspecies infancy GB-1496 strain has an antioxidant activity effect, and can reduce the concentration of free radicals so as to inhibit organ aging.
According to a specific embodiment of the present invention, preferably, in the above composition, the ratio of viable count of bifidobacterium animalis BB12 to bifidobacterium longum subspecies infancy GB-1496 is between 1:10 and 10:1, a step of; preferably 1:1, a step of; alternatively, the ratio of colony numbers of bifidobacterium animalis BB12 and bifidobacterium longum subspecies infantis GB-1496 is 1:10-10:1, a step of; preferably 1:1.
according to a specific embodiment of the present invention, preferably, in the above composition, the ratio of galactooligosaccharide, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 is 1g: (10 6 -10 13 )cfu：(10 6 -10 13 ) cfu; wherein the amounts of Bifidobacterium animalis BB12 and Bifidobacterium longum subspecies infantis GB-1496 are based on colony count or viable count.
According to a specific embodiment of the present invention, preferably, the content of the galactooligosaccharide in the above composition is 0.01 to 50g.
The composition of the present invention may be prepared by directly mixing the galacto-oligosaccharide-containing bacteria with the probiotic bacteria.
The invention also provides a nutritional composition comprising the above composition comprising galactooligosaccharides and probiotics. The nutritional composition may also contain other food or medicine allowable raw materials, adjuvants, and additives. In preparing the nutritional composition, the composition containing galactooligosaccharides and probiotics can be directly mixed with other raw materials, auxiliary materials and additives.
The invention also provides a food product comprising the above composition comprising galactooligosaccharides and probiotics and/or the above nutritional composition. The food can be infant food or senile food. According to the specific embodiment of the invention, the food can comprise one or more than two of milk powder, complementary food, beverage and the like, and can also contain other raw materials, auxiliary materials and additives which are allowed to be added.
The invention also provides the application of the combination of galactooligosaccharide, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 in promoting proliferation of the bifidobacterium subspecies infancy GB-1496.
The invention also provides application of the composition containing the galactooligosaccharide and the probiotics in promoting proliferation of bifidobacterium longum subspecies baby GB-1496.
According to a particular embodiment of the invention, the above promotion is preferably intended to promote the proliferation of bifidobacterium longum subspecies infantis GB-1496 in the gut.
The invention also provides the application of the combination of galactooligosaccharide, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 in promoting proliferation of the bifidobacterium subspecies infancy GB-1496; preferably in promoting colonisation and proliferation of bifidobacterium longum subspecies infantis GB-1496 in the gut.
According to a specific embodiment of the present invention, in the several applications mentioned above, the ratio of galactooligosaccharides, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 is preferably 1g: (10 6 -10 13 )cfu：(10 6 -10 13 ) cfu; wherein the amounts of Bifidobacterium animalis BB12 and Bifidobacterium longum subspecies infantis GB-1496 are based on colony count or viable count.
Bifidobacterium infantis is one of the earliest established bifidobacteria species in the infant's intestinal tract. Finding an effective method for promoting proliferation and dominant proliferation of bifidobacterium infantis, and is very important for intestinal colonization and further exerting other effects.
According to the invention, the research discovers that the compounded bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 can obviously promote the proliferation of bifidobacterium infancy, and the number of the proliferation is obviously higher than that of single combination of the GOS and the bifidobacterium animalis BB12 or the bifidobacterium longum subspecies infancy GB-1496. Moreover, in case a combination of GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 is used, bifidobacterium subspecies infancy GB-1496 exhibits advantageous proliferation.
Drawings
FIG. 1 shows the results of analysis of the significance of differences (OD 600 values) in promoting bifidobacterium proliferation when cultured with GOS as the carbon source, BB12, GB-1496 alone, and a combination of both.
FIG. 2 shows the results of PCR detection when GOS was used as a carbon source and BB12, GB-1496 alone and a combination of both were used for cultivation.
Microbial preservation for patent procedures:
GB-1496 Strain of the present invention
Preservation date: 10/04/2011
Preservation unit: china Center for Type Culture Collection (CCTCC)
Deposit unit address: 430072 of university of Wuhan, china
Preservation number: cctccc NO: m2011122
Classification naming: bifidobacterium longum subspecies infantis (Bifidobacterium longum subsp. Infantis)
Detailed Description
The technical solution of the present invention will be described in detail below for a clearer understanding of technical features, objects and advantageous effects of the present invention, but should not be construed as limiting the scope of the present invention.
The bifidobacterium longum subspecies of babies GB-1496 is human breast milk, and is preserved in China Center for Type Culture Collection (CCTCC), and the preservation unit address is: 430072, university of martial arts, chinese; preservation date: 04 th 2011; preservation number: cctccc NO: m2011122, classification nomenclature: bifidobacterium longum subspecies infantis (Bifidobacterium longum subsp.infantis).
The taxonomic characteristics of the strains were confirmed based on the 16S rDNA sequence analysis and the API bacteria identification system analysis results. The morphological and general characteristics of Bifidobacterium longum subspecies infant GB-1496 are detailed in Table 1.
TABLE 1
The bifidobacterium longum subspecies infantis GB-1496 strain was maintained at-80℃in MRS medium containing 20% glycerol. Before use, the mixture was activated twice at 37℃with MRS broth (DIFCO) containing 0.05% L-cysteine (24 hours).
Experimental example
(1) Preparing MRS basal medium: (1) weighing: weighing beef powder 5.0g, tryptone 10.0g, yeast powder 4.0g, tween 80 1.0mL, K 2 HPO 4 2.0g、CH 3 COONa 5.0g、C 6 H 5 O 7 (NH 4 )3 2.0g、MgSO 4 0.2g、MnSO 4 0.05g, 15.0g of agar powder and 1000mL of distilled water. (2) Mixing evenly: the weighed substances are uniformly mixed, heated and uniformly stirred. (3) Adjusting pH: with 1 mol/mL -1 NaOH adjusts the pH of the culture medium to 7.0-7.2. (4) Dissolving: pouring the dissolved culture medium into a triangular flask, adding a certain volume of water, and heating for dissolution. (5) And (5) subpackaging: pouring the prepared culture solution into a 250mL triangular flask, and wrapping the flask mouth with newspaper. (6) And (3) sterilization: the culture medium is sterilized by high pressure steam for 15min and then cooled for standby.
(2) On the premise of the same initial culture amount, the growth curve of GB-1496, BB12 and the compound bacteria (GB-1496:BB12=1:1) in the GOS carbon source is compared with the growth curve of the compound bacteria in a MRS+0.05% cysteine culture medium containing 1wt% of GOS. The results show that when GOS is used as a single carbon source, the combination of GB-1496 and BB12 can synergistically promote the increase of the total bifidobacteria, and the three groups have obvious differences in 24 hours. The specific results are shown in fig. 1 and table 2, wherein in fig. 1, # represents the difference from BB12 alone, # represents the difference from GB-1496 alone, and the combination of GB-1496 and BB12, vs GB-1496: p <0.01; combination of GB-1496 and BB12 vs BB12# #: p <0.01.
As can be seen from fig. 1: in the case of GOS as the carbon source, the combination of GB-1496 and BB12 is adopted, and the proliferation amount of the total bifidobacterium is obviously higher than that of the single GB-1496 and BB12.
TABLE 2
As can be seen from the data in table 2: the proliferation amount obtained by using the mixed strain is 1.5 times of the sum of BB12 and GB-1496 when cultured independently under the premise of the same culture initiation amount, and the unexpected improvement is obtained.
(3) PCR detection
In 24h, the proportion of two bacteria in the compound bacteria is detected by a PCR method, and the primer design is based on a 16SrRNA sequence, so that bifidobacterium animalis and bifidobacterium infantis can be distinguished specifically.
Wherein, the primers for specific differentiation of bifidobacterium lactis are as follows:
Bani-tF：TCACGACAAGTGGGTTGCCA(SEQ ID NO：1)
Bani-tR：GTTGATCGGCAGCTTGCCG(SEQ ID NO：2)。
primers specific for differentiating bifidobacterium longum subspecies infantis are as follows:
Binf-tF：ATCCGTCCGACCCAGACC(SEQ ID NO：3)
Binf-tR：CTCGACATCCTCACGGCC(SEQ ID NO：4)。
the results are shown in FIG. 2. As can be seen from fig. 2: in the mixed bacteria co-cultured by GB-1496 and BB12, bifidobacterium longum subspecies GB-1496 grow predominantly.
Example 1
The present example provides a composition comprising GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 comprising GOS, bifidobacterium animalis BB12, bifidobacterium longum subspecies infancy GB-1496, wherein GB-1496: BB12 = 1:1.
example 2
This example provides a set of nutritional compositions comprising the composition provided in example 1 comprising GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496. The nutritional composition has remarkable promoting effect on proliferation of bifidobacterium longum subspecies of infants GB-1496 in intestinal tracts, and can enable the bifidobacterium longum subspecies of infants GB-1496 to exhibit dominant growth.
Example 3
This example provides a group of pediatric foods comprising the composition comprising GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 provided in example 1, and one or a combination of two of the nutritional compositions provided in example 2, respectively, wherein the composition comprising GOS, bifidobacterium animalis BB12 and bifidobacterium subspecies infancy GB-1496, and the nutritional composition may be added alone, or may be added in any combination and in any ratio. The food can be infant food or senile food, such as milk powder, beverage, infant complementary food, etc.
The food has remarkable promoting effect on proliferation of Bifidobacterium longum subspecies baby GB-1496 in intestinal tract, and can promote growth of Bifidobacterium longum subspecies baby GB-1496.
Example 4
This example provides a group of medicaments comprising one or a combination of two of the composition comprising GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 provided in example 1 and the nutritional composition provided in example 2, respectively, wherein the composition comprising GOS, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 and the nutritional composition may be added individually or in any combination and in any ratio.
The medicine has remarkable promoting effect on proliferation of Bifidobacterium longum subspecies baby GB-1496 in intestinal tract, and can promote growth of Bifidobacterium longum subspecies baby GB-1496.
Sequence listing
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Claims (12)

1. A composition comprising galactooligosaccharides and probiotics, wherein the probiotics are bifidobacterium animalis BB12 and bifidobacterium longum subspecies infantis GB-1496;
wherein the ratio of the galactooligosaccharide, the bifidobacterium animalis BB12 and the bifidobacterium longum subspecies infancy GB-1496 is 1g: (10 6 -10 13 )cfu：(10 6 -10 13 )cfu。
2. The composition of claim 1, wherein the ratio of viable count of bifidobacterium animalis BB12 to bifidobacterium longum subspecies infancy GB-1496 is 1:10-10:1.
3. the composition of claim 2, wherein the ratio of viable count of bifidobacterium animalis BB12 to bifidobacterium longum subspecies infancy GB-1496 is 1:1.
4. composition according to claim 1 or 2, in which the galacto-oligosaccharide is present in an amount of 0.01-50g.
5. A nutritional composition comprising the composition according to any one of claims 1-4.
6. A food product comprising the composition of any one of claims 1-4 or the nutritional composition of claim 5.
7. The food according to claim 6, wherein the food is a pediatric food or an geriatric food.
8. The food according to claim 6, wherein the food comprises one or a combination of two or more of milk powder, a complementary food, and a beverage.
9. A medicament comprising the composition of any one of claims 1-4 or the nutritional composition of claim 5.
10. Use of a combination of galactooligosaccharide, bifidobacterium animalis BB12 and bifidobacterium longum subspecies infancy GB-1496 for promoting proliferation of bifidobacterium subspecies infancy GB-1496, wherein the ratio of galactooligosaccharide, bifidobacterium animalis BB12 and bifidobacterium subspecies infancy GB-1496 is 1g: (10 6 -10 13 )cfu：(10 6 -10 13 )cfu。
11. Use of a composition comprising galactooligosaccharides and probiotics as claimed in claim 1 for promoting proliferation of bifidobacterium longum subspecies infancy GB-1496.
12. Use according to claim 10 or 11, wherein said promotion is in promoting proliferation of bifidobacterium longum subspecies infantis GB-1496 in the gut.
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CN107212415A (en) * 2017-05-18 2017-09-29 江西长通实业有限公司 A kind of 5-linked probiotics nutrient powder and preparation method thereof
CN111067104A (en) * 2019-12-30 2020-04-28 黑龙江省完达山乳业股份有限公司 Composite probiotic infant formula milk powder and preparation method thereof

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CN107212415A (en) * 2017-05-18 2017-09-29 江西长通实业有限公司 A kind of 5-linked probiotics nutrient powder and preparation method thereof
CN111067104A (en) * 2019-12-30 2020-04-28 黑龙江省完达山乳业股份有限公司 Composite probiotic infant formula milk powder and preparation method thereof

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