CN113862311A

CN113862311A - Application of combination of isomerized lactose and 2' -fucosyllactose in promoting acid production of Bifidobacterium longum infant subspecies metabolism

Info

Publication number: CN113862311A
Application number: CN202111069426.9A
Authority: CN
Inventors: 刘彪; 石羽杰
Original assignee: Inner Mongolia Yili Industrial Group Co Ltd
Current assignee: Inner Mongolia Yili Industrial Group Co Ltd
Priority date: 2020-09-14
Filing date: 2021-09-13
Publication date: 2021-12-31

Abstract

The invention provides application of combination of isomerized lactose and 2' -fucosyllactose in promoting acid production of Bifidobacterium longum subspecies infantile metabolism. The invention also provides compositions, nutritional compositions, and food products comprising the combination of isomerized lactose and 2' -fucosyllactose. The bifidobacterium infantis is the earliest colonizing bacterium in the intestinal tract of an infant, and the search for effective prebiotics synergistic combination for promoting the metabolism and acid production of bifidobacterium longum subspecies YLGB-1496 is very important for the intestinal colonizing of the bifidobacterium longum subspecies YLGB-1496 and further playing other effects. The research of the invention finds that 2' FL and LOS can obviously promote the metabolism of Bifidobacterium longum subspecies YLGB-1496 to produce short chain fatty acid, especially lactic acid, under the specific combination proportion, which is obviously higher than the single prebiotics with the same amount and obviously higher than the single prebiotics with the same amount.

Description

Application of combination of isomerized lactose and 2' -fucosyllactose in promoting acid production of Bifidobacterium longum infant subspecies metabolism

Technical Field

The invention relates to application of a combination of isomerized lactose and 2' -fucosyllactose in promoting acid production by Bifidobacterium longum subspecies of infants YLGB-1496 metabolism, belonging to the technical field of health-care food.

Background

The intestinal system of the human body contains a large amount of bacteria, the total amount of which is more than 100 times of the cells of the human body, and the bacteria form the intestinal microecology of the human body. More and more studies indicate that the intestinal micro-ecology is closely related to the disease health of the human body, and especially the intestinal micro-ecology in the early life has an important influence on the health of life [1 ].

Previous studies have shown that different feeding regimes have a significant impact on the intestinal micro-ecology in infants in early stages. The ratio of bifidobacteria to lactobacilli in the faeces of the infants in the formula-fed group was significantly lower than that of the infants in the breast-fed group. By adding probiotics and prebiotics to infant formula milk powder, the intestinal micro-ecology of infants can be regulated, which is close to the intestinal micro-ecology constitution of breast-fed infants, especially the proportion of bifidobacteria [2 ].

Prebiotics are a class of carbohydrates that are not digestible by the human body and interact with the intestinal flora in the host's gut to produce substances or effects that are beneficial to the host [3 ]. Currently, most studied prebiotics include: galacto-oligosaccharides, fructo-oligosaccharides, inulin, isomerized lactose, polydextrose, xylo-oligosaccharides, and the like.

The isomerized lactose, of which the chemical name is 4-O-beta-D-galactopyranosyl-D-fructose, is a product prepared by alkali or non-alkali isomerization of lactose under the heating condition, is an isomer of lactose, and has reported health efficacy in a large number of clinical studies. First, isomerized lactose has a very good prebiotic effect, although it is not a dietary fiber. As early as 1957, Petuely found that isomerized lactose had a significant promoting effect on the growth of bifidobacteria [9], later studies found that the colonic flora composition of bottle-fed infants was very similar to that of adults, whereas if isomerized lactose was added to the formula, the composition of these infants was the same as breast-fed infants [54 ]. Subsequently in 1964, Hoffman et al studied the bacterial metabolism of isomerized lactose in the gut, demonstrating that it is a bifidogenic factor [10 ]. Later multiple clinical trials have demonstrated that isomerized lactose can act as a prebiotic for the adult population [55-57 ]. In vitro studies by Waston et al in 2013 found that many commercial probiotics could utilize isomerized lactose as a carbon source for growth [12 ]. Second, the use of isomerized lactose for the treatment of constipation dates back to the end of the last 60 years, and the first double-blind trial in 1968 reported that isomerized lactose could treat chronic constipation [47 ]. Recent clinical studies found that isomerized lactose (3.3 g/day) improved chronic constipation symptoms in children in china, including frequency of bowel movements and stool hardness, abdominal discomfort and flatulence, compared to the control group [48 ]. The 20ml x 2 isomerized lactose taken by adolescent people with chronic constipation can significantly increase defecation frequency with lower recurrence rate [49 ]. Isomerized lactose also has a clear effect on constipation problems of pregnant women [50-51] and elderly people [52], and the effects of isomerized lactose powder and syrup products are not significantly different [53 ]. Third, isomerized lactose is the only prebiotic currently reported to reduce the tendency to prevent necrotizing enterocolitis in preterm infants. A clinical study of preterm infants was conducted in 2010 by Riskin et al [11], and the experimental results showed that the addition of 1% (1g per 100mL of feed) of isomerized lactose to preterm infants was beneficial in reducing intolerance to enteral feeding, enabling preterm infants to be discharged earlier and home, and tended to have a lower incidence of necrotizing enterocolitis. Fourth, isomerized lactose aids in mineral absorption. In healthy adult males, supplemented with 300mg calcium, 150mg magnesium and isomerized lactose (0 g/day, 2 g/day, 4 g/day), respectively, urine was analyzed for calcium and magnesium, and the results showed that isomerized lactose improved the absorption of calcium and magnesium. But isomerized lactose did not affect the bone resorption marker in urine [58 ]. In climacteric women, breakfast for 9 consecutive days was supplemented with 5g and 10g of isomerized lactose, respectively, and the results showed that isomerized lactose improved calcium absorption and was dose dependent, with better effect in the high dose group [59 ]. Fifth, isomerized lactose can treat hepatic encephalopathy. An article in 1959 that found isomerized lactose to be a bifidus factor mentions that isomerized lactose may be useful in the treatment of hepatic encephalopathy [60 ]. Clinical studies confirm the effectiveness of isomerized lactose in treating hepatic encephalopathy. Later, instead of the standard treatment at that time, therapeutic doses for Hepatic Encephalopathy (HE) ranged up to 90 grams per day, depending on severity [61 ]. 30-60ml of isomerized lactose per day can improve cognitive function and health-related quality of life in patients with hepatic encephalopathy [62 ]. Sixth, isomerized lactose is not degraded by intestinal digestive juices and has a portion that permeates directly through the intestinal wall, so it is also routinely used clinically to assess intestinal permeability, safe and harmless [63 ]. Clinical observations have shown that the risk of peritonitis is lower in people taking isomerized lactose [64], and this result has written the prevention and treatment recommendations for ISPD peritonitis [65 ]. Seventh, isomerized lactose can reduce fat. In middle-aged women, 300mg calcium, 150mg magnesium and 4g isomerized lactose were supplemented, and at 12 months, body fat was lower than that of the control group [66 ]. Finally, there have been several studies reporting clinical studies on isomerized lactose to lower blood lipids [67-69], prevent infantile eczema [70], prevent female vaginitis [71-72], and treat (non-typhoid) salmonella carriers [73 ].

The breast milk naturally contains more than 200 kinds of oligosaccharides, the total concentration of the oligosaccharides is higher than the concentration of proteins in the breast milk, and the oligosaccharides are substances which cannot be directly absorbed by human bodies, but play an important role in establishing intestinal flora in the early life stage, and become hot spots of research in recent years. HMOs in human breast milk consist of 5 monomers: d-glucose (Glc), D-galactose (Gal), N-acetylglucosamine (GlcNAc), L-fucose (Fuc) and sialic acid (Sia). These 5 monomers, combined in different proportions, constitute thousands of HMOs in breast milk [2 ]. HMOs can be classified into neutral human milk oligosaccharides and acidic human milk oligosaccharides. Neutral human milk oligosaccharides are oligosaccharides comprising fucosyl, acidic human milk oligosaccharides are oligosaccharides comprising sialic acid and its sulfate structure. The neutral human milk oligosaccharide accounts for about 70% of human milk, and the acidic human milk oligosaccharide accounts for about 30% of human milk. Only about 200 HMOs molecules have been identified in breast milk, with neutral fucosylated HMOs being the predominant species. While the oligosaccharides in cow's and goat's milk are mainly acidic sialylated HMOs. 2' -Fucosyllactose (2 ' Fucosyllactose, 2' FL) is one of the major neutral HMOs molecules in breast milk, a uniquely abundant substance in human milk [6], which is deficient in traditional infant food, especially in infant formula products. 2' FL is a neutral HMO and is one of the most concentrated oligosaccharides in breast milk. Compared with the milk of other mammals transversely, the common livestock milk contains almost no 2' FL, can not be detected in cow milk and horse milk, and has extremely low concentration in sheep milk and goat milk [74 ]; only a few primates and zoo mammals have higher concentrations of 2'FL in their milk, and analysis of the milk from 13 mammals by Warren et al shows that the 2' FL from gorilla milk is closest to breast milk at a concentration of about 56% of breast milk, followed by a large gray bear at about 44% of breast milk. The 2'FL concentration in the milk of black bear and black rhinoceros is about 12% of that of breast milk, and the 2' FL concentration in the milk of giraffe is about 4% of that of breast milk; the dog's milk does not contain 2' FL [75], and the monkey's milk does not contain 2' FL [76 ].

Existing research supports the safety and tolerability of 2' FL. 2' FL intervention experiments (29.30-298.99mg/kg/day) were carried out for 3 weeks in the piglet model with no adverse effects [78 ]. 2' FL (0-6000 mg/kg bw/day) was evaluated in the adolescent rat model and no adverse reaction was observed [77 ]. Reverri EJ et al reviewed the relevant clinical studies currently being conducted, and considered that the addition of 2'FL to infant formula is safe and metabolically similar to 2' FL in breast milk [79 ].

The health efficacy of 2' FL is related to many aspects. First, 2' FL has a significant effect on the composition of the intestinal flora after birth of the infant. Avershina et al report that after birth, bifidobacteria gradually become the dominant bacteria in the infant gut, reaching a peak at 4 months after birth, accounting for 60% of total bacterial load, while adults are only about 2% [80 ]. Bunesova et al isolated various Bifidobacterium strains from the Kenya infant gut, whereas only the genomic sequences of Bifidobacterium infantis, Bifidobacterium bifidum, etc, containing alpha-fucosidases (alpha-fucosidases) were converted to 1,2-PD by 2' FL for use by gut cells [81 ]. 2' FL inhibits intestinal deleterious bacteria, and enterobacteria closely associated with necrotizing enterocolitis cannot consume 2' FL, 6-sialyllactose (6 ' -SL) and LNnT for growth, but can grow using Galactooligosaccharides (GOS) [82 ]. Supplementation of 2' FL after cecostomy in an animal model of short bowel syndrome can promote weight gain and adaptive histological changes of intestinal mucosa [83] and improve symptoms.

Second, 2' FL has anti-infective activity. Breast-fed infants have a lower incidence of infectious diseases than formula-fed infants. Clinical studies by Morrow et al found that a higher total amount of breast-milk alpha-1, 2 fucosylated HMOs (including 2' FL) in breast-fed infants was significantly negatively correlated with diarrhea caused by campylobacter jejuni infection [84 ]. In vitro studies find that 2'FL can inhibit the adhesion of Campylobacter jejuni, enteropathogenic Escherichia coli, Salmonella, Pseudomonas aeruginosa and intestinal epidermal cells, and 2' FL can also inhibit the adhesion of Pseudomonas aeruginosa and respiratory epithelial cells [85 ]. The effect of 2'FL on treatment of acute intestinal infections and immune responses was evaluated in a mouse model fed antibiotic followed by jejunum, and the results showed that ingestion of 2' FL reduced the colonization of jejunum and induction of mucosal immunoreactive inflammatory signaling molecules in the acute phase by 50-60% [86 ]. In vitro experiments have shown that 2'FL and 3' FL compete for norovirus inhibition to reduce the risk of infection [87 ]. 2' FL can reduce respiratory syncytial virus infection respiratory epithelial cell obviously and regulate the expression of cell factor and inflammation factor [88 ].

Third, 2' FL has the function of regulating immunity. After Marriage et al recruited 420 healthy term infants 5d after birth, randomly divided into 4 groups fed with different milk powders, namely a control group (2.4g/LGOS), a low dose 2'FL group (0.2 g/L2' FL +2.2g/L GOS), a high dose 2'FL group (1 g/L2' FL +1.4g/LGOS) and a breast-feeding control group, the infants in the four groups showed no difference in growth after 4 months of feeding. The absorption profile of 2'FL was similar for breast-feeding and for the formula group of 2' FL [89 ]. Goehring et al detected inflammatory factors in the blood of the tested infants at 6w and found that the inflammatory factors IL-1, IL-1 alpha, IL-1 beta, IL-6 and TNF-alpha were significantly elevated in the control group of plain infant formula [90 ].

Gram-negative bacteria cause intestinal infections that are triggered mostly by lipopolysaccharide (lipopolysaccharide) in combination with TLR4 of intestinal epidermal cells to trigger inflammation. 2'FL can weaken the induction expression of intestinal IL-8 and reduce the inflammatory response of intestinal mucosa cells by inhibiting the mRNA transcription of TLR4 upstream regulatory factor CD14 and the translation and transportation of CD14, and the effect is close to that of human milk HMO [91] containing 2' FL at an equal concentration. DC-SIGN, which is the major antigen presenting cell on the surface of the intestinal tract, also binds to 2' FL, and is presumed to be one of the mechanisms for modulating immune activity [92 ].

Fourth, 2' FL can affect cognitive function. After 5 weeks of feeding 350mg of 2' -FL/kg, the long-range enhancing effect of the hippocampus of the rodent can be detected to be enhanced, and the corresponding cognitive behavior experiment shows better. Increased protein expression in the brain of animals in the 2' FL group associated with newly acquired memory stores [93 ]. Rats were fed supplemental 1g/kg of 2'FL during lactation and the 2' FL group performed slightly better in the Morris water maze experiment after weaning (4-6 weeks). By age 1, adult rats in the 2' FL group still performed better in the new object recognition and Y maze model. Thus, 2' -FL has both short-term and long-term effects, and the intensity and duration of brain LTP are higher than those of control [94 ]. In vitro studies have also shown that L-fucose and 2' FL increase the electrophysiological signal index enhancement in the hippocampus of rats [95 ]. Further animal experiments found that the gut-brain axis plays a key role in 2 'FL-mediated cognitive benefits, and vagotomy inhibits the effects of oral 2' FL on LTP and learning cognition [96 ].

Fifth, 2' FL can affect allergy. Castillo-Courtade et al assessed that mice fed 2'FL had a milder response, a lower number of intestinal mast cells, and a CD4+ CD25+ IL-10+ cell number in the Peyer's lymph node in an ovalbumin-stimulated mouse food allergy model [97 ].

The breast milk not only contains rich oligosaccharide substances, but also contains live microbial flora. For breast-fed infants, breast milk microecology is a key contributing factor to the development of intestinal microecology in infants. The most abundant intestinal micro-ecology of breast-fed infants in the early stages of life is bifidobacteria. Further microbiological analysis tests show that the intestinal flora of breast-fed infants is different from that of adults and is rich in bifidobacterium infantis. Studies have shown that the efficacy of bifidobacterium infantis mainly includes the following aspects:

firstly, bifidobacterium infantis has strong capacity of utilizing HMO and has advantages in intestinal colonization, and unlike other bifidobacteria, the genome of bifidobacterium infantis contains more extensive genes [13-19] related to HMO transportation and metabolism, and in vitro studies, bifidobacterium infantis grows better than other strains in the presence of HMO [19,25], so the bifidobacterium infantis strain has unique advantages over other conventional probiotics. Bifidobacterium infantis is also able to metabolize the short-chain fatty acids produced which reduce intestinal pathogens and promote the growth and maturation of intestinal epidermal cells [36 ]. Short chain fatty acids, commonly known as acetic, propionic, butyric and lactic acids, have various health benefits in addition to being absorbed as energy substances by intestinal cells, which is beneficial to the integrity of intestinal epidermal cells [42 ]. Among them, lactic acid has a variety of metabolic and regulatory properties, and L-lactic acid is a natural ligand of gastrointestinal coupled GPR81, inhibiting cAMP mediated intracellular signaling events such as lipolysis [38 ]. Lactic acid can also be converted to propionic acid or butyric acid by the action of bacteria using lactic acid [39 ]. Innovations in certain sports nutrition focus on novel probiotics, achieve conversion of lactic acid to propionic acid butyric acid, and improve exercise endurance and performance [40 ]. Animal experiments have shown that L-lactic acid stimulates hippocampal nerve growth in adult mice [41 ].

Secondly, bifidobacterium infantis has an anti-inflammatory effect, intestinal inflammation of premature infants is an important cause of reducing Necrotizing Enterocolitis (NEC) of premature infants. Research on intestinal cells as a model shows that the bifidobacterium infantis can induce the intestinal cells to express anti-inflammatory cytokines [20-23], and rat animal experiments show that the addition of the bifidobacterium infantis can reduce the inflammation of NEC [24 ]. Bifidobacteria are often deficient in the gut of preterm infants [26] and the incidence of NEC in breast-fed preterm infants is significantly lower than in formula-fed infants [27 ]. Unlike the breast milk of term infants, the breast milk of preterm infants contains a higher concentration of fucosylated HMO [28 ]. Comparing the feeding effect of bifidobacterium infantis and bifidobacterium lactis added in the premature infant population, the result shows that the colonization ability of bifidobacterium infantis is stronger than that of bifidobacterium lactis no matter whether the formula is fed or breast-fed, and the group of the breast-fed infants added with bifidobacterium infantis is the group with the highest number of bifidobacterium faecalis [29 ]. A number of clinical studies have shown that the risk of NEC development can be reduced by the administration of Bifidobacterium infantis to preterm infants [31-35 ].

Finally, the advantage of adding bifidobacterium infantis is also reflected in the aspect of immunity, and in a full-term infant clinical study of Bengal, the number of bifidobacterium infantis in the intestinal tract of an infant is found to be positively correlated with the vaccine response in the early four months of life, namely the vaccine response has a higher index [30-31] when the number of bifidobacterium infantis is higher, and the trend still exists [31] in 2 years after birth, and the bifidobacterium infantis is supposed to be helpful for activating the immune function and the immunological memory function.

However, despite the above-mentioned multifaceted functions of bifidobacterium infantis, there is still a lack in the art of an infant food product effective in promoting the metabolism of bifidobacterium infantis to produce acid.

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Disclosure of Invention

In order to solve the above technical problems, the present invention aims to provide the use of a combination of isomerized lactose and 2' -fucosyllactose for promoting the metabolism of bifidobacterium longum infantis ylb-1496 for producing acid.

It is also an object of the present invention to provide a composition, a nutritional composition and a food product comprising isomerized Lactose (LOS) and 2 '-fucosyllactose (2' FL).

To achieve the above objects, the present invention provides the use of a combination of LOS and 2' FL for promoting the metabolic production of acid by Bifidobacterium longum subspecies of infantis YLGB-1496. The combination of LOS and 2' FL is capable of significantly promoting the metabolic acidogenesis of Bifidobacterium longum subspecies of infants YLGB-1496.

According to a particular embodiment of the present invention, preferably, the acid is a short chain fatty acid; more preferably, the short chain fatty acid is lactic acid.

According to the specific embodiment of the present invention, preferably, the mass ratio of the LOS to the 2' FL is 1: 10-10: 1, e.g. 10: 1. 4: 1. 1: 1. 1: 4 or 1: 10. and controlling the mass ratio of LOS to 2' FL to be 1: 10-10: 1, the bifidobacterium longum infantis YLGB-1496 produces acid when grown at a higher concentration of short chain fatty acids than when grown in medium containing 2'FL alone, wherein the lactic acid concentration is significantly higher than when grown in medium containing 2' FL alone, and also higher than when grown in medium containing LOS alone.

According to the specific embodiment of the present invention, preferably, the mass ratio of the LOS to the 2' FL is 1: 4. and controlling the mass ratio of LOS to 2' FL to be 1: 4, the concentration of short chain fatty acids, lactic acid produced by bifidobacterium longum infantis YLGB-1496 when grown is higher than its corresponding concentration when grown in medium containing LOS alone and significantly higher than its corresponding concentration when grown in medium containing 2' FL alone.

According to the specific embodiment of the present invention, preferably, the mass ratio of the LOS to the 2' FL is 1: 10-4: 1; more preferably 1: 1-4: 1. and controlling the mass ratio of LOS to 2' FL to be 1: 10-4: 1, the lactic acid concentration produced by bifidobacterium longum infantis YLGB-1496 when grown is higher than that corresponding to growth in medium containing 2' FL alone and also higher than that corresponding to growth in medium containing LOS alone. Wherein, compared with a culture medium containing LOS alone, when the mass ratio of LOS to 2' FL is 1: 1-4: 1, the difference in lactic acid concentration between the two was higher.

According to the specific embodiment of the present invention, preferably, the mass ratio of the LOS to the 2' FL is 4: 1. in the following step 4: 1, the concentration of short-chain fatty acid and the concentration of lactic acid produced by the bifidobacterium longum subspecies YLGB-1496 during growth are both obviously higher than the corresponding concentration of short-chain fatty acid and lactic acid when the bifidobacterium longum subspecies YLGB-1496 is grown in a culture medium containing 2' FL alone, and simultaneously, the concentration of lactic acid is also obviously higher than the concentration of lactic acid when the bifidobacterium longum subspecies YLGB-1496 is grown in a culture medium containing LOS alone.

According to the specific embodiment of the invention, the mass ratio of LOS to 2' FL is 1: 10-10: OD of Bifidobacterium longum subspecies YLGB-1496 growth in media within the range of 1 (e.g., 1: 10, 1: 4, 1: 1, 4: 1, or 10: 1)₆₀₀The value is significantly higher than that of the singleOD grown in 2' FL-only medium₆₀₀At the same time, bifidobacterium longum subspecies infants YLGB-1496 grew at a pH significantly lower than it grew in medium containing 2' FL alone.

According to a specific embodiment of the present invention, preferably, in the above prebiotic composition, the mass ratio of LOS to 2' FL is 1: 10-4: 1. more preferably 1: 4-4: 1, e.g. 1: 4 or 4: 1. the mass ratio of LOS to 2' FL is 1: 10-4: OD of growth of Bifidobacterium longum subspecies YLGB-1496 in medium within the range of 1₆₀₀The value is higher than the OD of the strain grown in the culture medium containing LOS alone₆₀₀The value is obtained. Compared with a culture medium containing LOS alone, when the mass ratio of LOS to 2' FL is 1: 4-4: 1 when both are at OD₆₀₀The significant difference in value is higher; in particular, in the following 1: 4. 4: 1 the most significant difference between these two ratios.

According to a specific embodiment of the present invention, preferably, in the above prebiotic composition, the mass ratio of LOS to 2' FL is 4: 1. and controlling the mass ratio of LOS to 2' FL at 4: 1, bifidobacterium longum subspecies infantis YLGB-1496 is grown at a pH lower than that of its growth in medium containing LOS alone, significantly lower than that of its growth in medium containing 2' FL alone; at the same time, its OD₆₀₀The values were significantly higher than their OD grown in medium with LOS alone and medium with 2' FL alone, respectively₆₀₀The value is obtained.

The invention also provides a composition containing LOS and 2'FL, preferably, in the composition, the mass ratio of the LOS to the 2' FL is 1: 10-10: 1.

according to a specific embodiment of the present invention, wherein, in the above composition, the mass ratio of LOS to 2' FL may be 1: 10-4: 1, preferably 10: 1. 1: 4. 1: 1 or 4: 1.

according to a specific embodiment of the present invention, preferably, in the above composition, the mass ratio of LOS to 2' FL is 4: 1 or 1: 4.

according to a specific embodiment of the present invention, the specific mass ratio in the above range of the mass ratio of LOS to 2' FL may be 1: 10. 1: 4. 1: 1. 4: 1. 10: 1, etc., such as 1: 10-4: the mass ratio range of 1 includes specific mass ratios of 1: 10. 1: 4. 1: 1. 4: 1, etc.

According to a particular embodiment of the invention, preferably, the above composition further comprises bifidobacterium longum subsp.

According to a particular embodiment of the invention, in the composition of the invention, said Bifidobacterium longum infantis YLGB-1496 is Bifidobacterium longum infantis (Bifidobacterium longum subsp. The strain is preserved in China Center for Type Culture Collection (CCTCC) with the preservation unit address: wuhan university, Wuhan, China, 430072; the preservation date is as follows: 2011, 04 month 10 days; the preservation number is: CCTCC NO: m2011122, category naming: bifidobacterium longum subsp.

The bifidobacterium longum subspecies infantis CCTCC No. M2011122 has better free radical scavenging capacity and reducing capacity, and can induce Caco-2 cells to increase the expression of antioxidant enzymes, so the bifidobacterium longum subspecies infantis YLGB-1496 strain has the activity effect of resisting oxidation, and can reduce the concentration of free radicals to inhibit organ aging.

The composition provided by the invention can be prepared by directly mixing LOS and 2' FL, and when the composition contains Bifidobacterium longum subspecies YLGB-1496, the composition can be added by a conventional method.

The invention also provides a nutritional composition comprising the above composition comprising LOS and 2' FL. The nutritional composition may also contain other food or medicine raw materials, adjuvants, and additives. In preparing the nutritional composition, the composition containing LOS and 2' FL can be directly mixed with other raw materials, adjuvants, and additives.

The invention also provides a food product comprising the above composition comprising LOS and 2' FL and/or the above nutritional composition. The food can be food for children or food for the elderly. According to a specific embodiment of the present invention, the food may include one or a combination of two or more of milk powder, supplementary food, beverage, etc., and may further include other raw materials, auxiliary materials, and additives that are allowed to be added.

The invention also provides a medicament comprising the above composition comprising LOS and 2' FL and/or the above nutritional composition. The medicine may also contain other raw materials, adjuvants, and additives.

The invention also provides application of the composition containing LOS and 2' FL in promoting acid production by Bifidobacterium longum subspecies YLGB-1496 in metabolism, preferably in promoting acid production by Bifidobacterium longum subspecies YLGB-1496 in intestinal metabolism, and especially in promoting lactic acid production by Bifidobacterium longum subspecies YLGB-1496 in intestinal metabolism.

The invention also provides application of the nutritional composition in promoting acid production by metabolism of bifidobacterium longum subspecies YLGB-1496, preferably in promoting acid production by metabolism of bifidobacterium longum subspecies YLGB-1496 in intestinal tracts, and particularly in promoting lactic acid production by metabolism of bifidobacterium longum subspecies YLGB-1496 in intestinal tracts.

The bifidobacterium infantis is the earliest colonizing bacterium in the intestinal tract of an infant, and the search for effective prebiotics synergistic combination for promoting the metabolism and acid production of bifidobacterium longum subspecies YLGB-1496 is very important for the intestinal colonizing of the bifidobacterium longum subspecies YLGB-1496 and further playing other effects. The research of the invention finds that 2' FL and LOS can obviously promote the metabolism of Bifidobacterium longum subspecies YLGB-1496 to produce short chain fatty acid, especially lactic acid, under the specific combination proportion, which is obviously higher than that of single prebiotics with the same quantity.

Drawings

FIG. 1 shows the analysis of the significant difference between the combination of LOS and 2'FL in different mass ratios and the simple use of 2' FL and LOS in promoting the growth of Bifidobacterium longum subspecies of infants YLGB-1496.

FIG. 2 shows the analysis results of the significant difference of the combination of LOS and 2'FL with different mass ratios and the simple use of 2' FL and LOS in pH value.

FIG. 3 shows the analysis of the significant difference between the combination of LOS and 2'FL in different mass ratios and the simple use of 2' FL and LOS in promoting the metabolism of short chain fatty acids by Bifidobacterium longum subspecies of infant YLGB-1496.

FIG. 4 shows the analysis results of the significant difference between the combination of LOS and 2'FL with different mass ratios and the simple use of 2' FL and LOS in promoting the metabolism of Bifidobacterium longum subspecies YLGB-1496 for producing lactic acid.

Microbial deposits for patent procedures:

the YLGB-1496 strain of the invention (once named: GB-1496 strain)

The preservation date is as follows: 2011 10/04/month

The preservation unit: china Center for Type Culture Collection (CCTCC)

The address of the depository: wuhan university, Wuhan, China 430072

The preservation number is: CCTCC NO: m2011122

And (3) classification and naming: bifidobacterium longum subsp

Detailed Description

The technical solutions of the present invention will be described in detail below in order to clearly understand the technical features, objects, and advantages of the present invention, but the present invention is not limited to the practical scope of the present invention.

The bifidobacterium longum subspecies of infants YLGB-1496 is derived from human breast milk, is preserved in China Center for Type Culture Collection (CCTCC), and has the preservation unit address: wuhan university, Wuhan, China, 430072; the preservation date is as follows: 2011, 04 month 10 days; the preservation number is: CCTCC NO: m2011122, category naming: bifidobacterium longum subsp.

The taxonomical characteristics of the strain were confirmed based on the results of 16S rDNA sequence analysis and analysis by the API bacterial identification system. The morphological and general properties of Bifidobacterium longum subspecies of the infant YLGB-1496 are characterized in detail in Table 1.

TABLE 1

Bifidobacterium longum strain YLGB-1496 of subspecies infantis was maintained in MRS medium containing 20% glycerol at-80 ℃. Before use, the mixture was activated twice (24 hours) at 37 ℃ with MRS broth (DIFCO) containing 0.05% L-cysteine.

Examples of the experiments

Bifidobacterium longum subspecies-infantile YLGB-1496 was grown in 7 media of 1% MRS + 0.05% cysteine, respectively, and their growth and short chain fatty acid metabolism were determined. Wherein, the composition of MRS is shown in table 2.

TABLE 2

The results of significant difference analysis of experimental results are shown in fig. 1 and fig. 2, wherein, # denotes the difference from using 2' FL alone, # denotes the difference from using LOS alone, # denotes p < 0.001; # # # # represents p < 0.0001; represents p < 0.05; represents p < 0.01; represents p < 0.001.

As can be seen from table 2, fig. 1, and fig. 2: after 48 hours of culture, bifidobacterium longum subspecies infants strain YLGB-1496 were cultured in LOS: 2' FL ═ 1: 10. 1: 4. 1: 1. 4: 1. 10: 1, the growth conditions in the culture medium are better; wherein:

in LOS: 2' FL ═ 1: 10. 1: 4. 1: 1. 4: 1. 10: 1, YLGB-1496 corresponding OD₆₀₀The values are all significantly higher than the OD of the medium using 2' FL alone₆₀₀A value;

in LOS: 2' FL ═ 1: 10. 1: 4. 1: 1. 4: 1, YLGB-1496 corresponding OD₆₀₀The values are all higher than the OD of the culture medium using LOS alone₆₀₀Values, in particular between 1:4. 4: 1, the significant difference is higher;

the above results show that the combination of LOS and 2' FL has a certain synergistic effect on the growth of YLGB-1496.

Meanwhile, in LOS: 2' FL ═ 1: 10. 1: 4. 1: 1. 4: 1. 10: 1, the corresponding pH of YLGB-1496 was significantly lower than that grown in LOS and 2'FL containing media, indicating that the combination of LOS and 2' FL significantly promoted the metabolism of short chain fatty acids at the above ratios.

After 48 hours of culture, the content of short chain fatty acids was measured, as shown in Table 3.

The results of significant difference analysis of the results of detection of short chain fatty acids and lactic acid are shown in fig. 3 and 4, wherein # represents the difference from the simple use of 2' FL, # represents the difference from the simple use of LOS, # represents p < 0.001; # # # # represents p < 0.0001; represents p < 0.05; represents p < 0.01; represents p < 0.001.

TABLE 3

	Lactic acid	Acetic acid	Propionic acid	Total up to
					LOS	2.11±0.03	3.91±0.20	0.81±0.02	6.83±0.25
2′FL	1.08±0.01	3.83±0.03	0.84±0.03	5.76±0.08
					LOS/2′FL 1/10	2.25±0.01	3.88±0.09	0.86±0.03	7.00±0.13
LOS/2′FL 1/4	2.70±0.16	3.95±0.07	0.87±0.02	7.52±0.25
					LOS/2′FL 1/1	2.46±0.04	3.97±0.12	0.85±0.01	7.28±0.17
LOS/2′FL 4/1	2.44±0.05	3.88±0.05	0.84±0.01	7.16±0.11
					LOS/2′FL 10/1	2.41±0.12	3.95±0.06	0.86±0.01	7.22±0.19

As can be seen from table 3, fig. 4, bifidobacterium longum subspecies infants yggb-1496 were cultured for 48 hours in LOS: 2' FL ═ 1: 10. 1: 4. 1: 1. 4: 1. 10: 1, the acid production condition in the culture medium is better; wherein:

in LOS: 2' FL ═ 1: 10. 1: 4. 1: 1. 4: 1. 10: 1, the concentration of short-chain fatty acid and the concentration of lactic acid corresponding to YLGB-1496 are obviously higher than those of the medium which only uses 2' FL;

in LOS: 2' FL ═ 1: 1. 4: 1, the concentration of lactic acid corresponding to YLGB-1496 is obviously higher than that of the medium which only uses LOS; in LOS: 2' FL ═ 4: 1, the total amount of the short-chain fatty acid corresponding to YLGB-1496 is increased, and compared with LOS alone, 2'FL alone and the combination of 2' FL and LOS with other mass ratios, the total amount of the short-chain fatty acid is obviously different;

the above results show that the combination of LOS and 2' FL has a certain synergistic effect on acid production by YLGB-1496, and particularly significantly promotes the production of lactic acid.

Example 1

The present example provides a group of compositions capable of promoting the metabolic production of acid by bifidobacterium longum subsp. 10. 1: 4. 1: 1. 4: 1. 10: LOS of 1 and 2' FL.

Example 2

The present example provides a group of compositions capable of promoting the metabolic production of acid by bifidobacterium longum subsp. 10. 1: 4. 1: 1. 4: 1. 10: 1 LOS and 2' FL, and Bifidobacterium longum subspecies-infantis YLGB-1496.

Example 3

The embodiment provides a composition capable of promoting bifidobacterium longum subspecies infants YLGB-1496 to metabolize to produce lactic acid, which comprises the following components in a mass ratio of 1: LOS of 4 and 2' FL. The composition has short chain fatty acid production promoting effect on Bifidobacterium longum subspecies of infants YLGB-1496 superior to that of LOS alone, and has lactic acid production promoting effect on Bifidobacterium longum subspecies of infants YLGB-1496 significantly superior to that of LOS alone and 2' FL alone.

Example 4

The embodiment provides a composition capable of promoting bifidobacterium longum subspecies infants YLGB-1496 to metabolize to produce lactic acid, which comprises the following components in a mass ratio of 1: LOS of 1 and 2' FL. The composition has short chain fatty acid production promoting effect on Bifidobacterium longum subspecies of infants YLGB-1496 superior to that of 2'FL alone, and has lactic acid production promoting effect on Bifidobacterium longum subspecies of infants YLGB-1496 significantly superior to that of LOS alone and 2' FL alone.

Example 5

The embodiment provides a composition capable of promoting bifidobacterium longum subspecies infants YLGB-1496 to metabolize to produce lactic acid, which comprises the following components in a mass ratio of 4: LOS of 1 and 2' FL. The composition has a short chain fatty acid production promoting effect on Bifidobacterium longum subspecies of infants YLGB-1496 significantly better than 2'FL alone, and a lactic acid production promoting effect on Bifidobacterium longum subspecies of infants YLGB-1496 significantly better than LOS alone and 2' FL alone.

Example 6

This example provides a set of nutritional compositions, each containing a composition as provided in any one of examples 1-5.

Example 7

This example provides a set of pediatric foods comprising each of the compositions provided in any of examples 1-5 and/or each of the nutritional compositions provided in example 6, wherein the compositions and nutritional compositions may be added individually or in any combination and in any proportions. The food can be food for children or elderly people, such as milk powder, beverage, and infantile supplementary food.

Example 8

This example provides a set of drugs that each contain the compositions provided in any of examples 1-5 and/or the nutritional compositions provided in example 6, wherein the compositions and nutritional compositions can be added individually or in any combination and in any ratio.

Claims

1. Use of a combination of isomerized lactose and 2' -fucosyllactose to promote the metabolic acidogenesis of bifidobacterium longum subspecies of infants, YLGB-1496; preferably, the acid is a short chain fatty acid; more preferably, the short chain fatty acid is lactic acid.

2. The use according to claim 1, wherein the mass ratio of isomerized lactose to 2' -fucosyllactose is 1: 10-10: 1.

3. use according to claim 1 or 2, wherein the mass ratio of isomerized lactose to 2' -fucosyllactose is 1: 4.

4. use according to claim 1 or 2, wherein the mass ratio of isomerized lactose to 2' -fucosyllactose is 1: 10-4: 1; preferably 1: 1-4: 1.

5. the use according to claim 1,2 or 4, wherein the mass ratio of isomerized lactose to 2' -fucosyllactose is 4: 1.

6. a composition comprising isomerized lactose and 2 '-fucosyllactose, wherein the mass ratio of isomerized lactose to 2' -fucosyllactose is 1: 10-10: 1; preferably 1: 10. 1: 4. 1: 1. 4: 1 or 10: 1; more preferably 1: 4 or 4: 1;

preferably, the composition further comprises bifidobacterium longum subspecies of infantis YLGB-1496.

7. A nutritional composition comprising the isomerized lactose-with 2' -fucosyllactose-containing composition of claim 6.

8. A food product comprising the composition of claim 6 and/or the nutritional composition of claim 7;

preferably, the food is a pediatric food or an geriatric food;

preferably, the food comprises one or more of milk powder, supplementary food and beverage.

9. A medicament comprising the composition of claim 6 and/or the nutritional composition of claim 7.

10. Use of a composition according to claim 6 and/or a nutritional composition according to claim 7 for promoting metabolic acidogenesis of bifidobacterium longum subsp.

Preferably in promoting the metabolism of bifidobacterium longum subspecies of infants YLGB-1496 in the intestinal tract to produce acid;

more preferably in promoting the metabolism of bifidobacterium longum subspecies of infants YLGB-1496 in the intestinal tract to produce lactic acid.