CN106923345B - Stabilized probiotic compositions - Google Patents
Stabilized probiotic compositions Download PDFInfo
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- CN106923345B CN106923345B CN201710163295.8A CN201710163295A CN106923345B CN 106923345 B CN106923345 B CN 106923345B CN 201710163295 A CN201710163295 A CN 201710163295A CN 106923345 B CN106923345 B CN 106923345B
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- 229940046010 vitamin k Drugs 0.000 description 1
- 239000001717 vitis vinifera seed extract Substances 0.000 description 1
- 238000005550 wet granulation Methods 0.000 description 1
- 229940100445 wheat starch Drugs 0.000 description 1
- 235000008939 whole milk Nutrition 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 235000010930 zeaxanthin Nutrition 0.000 description 1
- 239000001775 zeaxanthin Substances 0.000 description 1
- 229940043269 zeaxanthin Drugs 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
- 235000021241 α-lactalbumin Nutrition 0.000 description 1
Classifications
-
- 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
Abstract
The present invention relates to stable probiotic compositions. In particular, the present invention relates to a probiotic composition comprising: probiotic bacteria, phospholipids, glycerides, optionally carbohydrates, and optionally binders, the probiotic composition being in the form of a granulate or powder and having a water content of less than 4%. The probiotic composition of the invention comprises 1 x 10 of probiotic per 1g6‑1×1014cfu of probiotic bacteria. Also relates to a method for preparing the probiotic composition and a nutritional composition comprising the probiotic composition. The probiotic composition of the present invention has excellent properties such as excellent stability.
Description
Technical Field
The present disclosure relates to stabilized compositions of biological material for ingestion by an individual. More specifically, the present disclosure relates to stabilized mixtures comprising at least one phospholipid and at least one glyceride that together provide improved stability to probiotic organisms when the probiotic is included in a nutritional composition. The present disclosure also includes a probiotic stabilization method.
Background
There are currently a wide variety of compositions used to supplement nutrition for both humans and animals. These supplements may be provided to alter, reduce or increase the microbiota in the gut of an individual, thereby producing a desired effect on digestion. Ideally, supplementation may be based on altering specific bacteria within the Gastrointestinal (GI) tract of a human body to culture an improved microbiota for an individual (including a human). This type of supplementation may be carried out by using probiotics, which are understood to be viable microorganisms that, when administered in effective amounts, confer health or nutritional benefits to the host. One of the more common types of probiotic bacteria is lactic acid bacteria, which are capable of converting sugars and other carbohydrates into lactic acid. This transformation lowers the pH in the intestinal tract of the host and provides less opportunity for harmful organisms to grow and cause problems through gastrointestinal infections.
A common technical challenge is to introduce probiotics into a host in a suitable manner for both maintenance of the probiotics and for the health and pleasure of the individual. Current techniques include the use of encapsulation and stabilization techniques to shield the probiotic with a protective layer or matrix so that the protected microorganisms can be delivered to the appropriate location within the GI tract of the individual.
Although there has been development of encapsulation and stabilization techniques involving microorganisms for delivery to the digestive system of animals, little has been developed in encapsulation or stabilization techniques that protect the viability of probiotics during distribution and storage. There is a need for useful stabilization techniques where refrigeration is excluded, and further where such formulations may be exposed to various environments, especially those associated with tropical climates. In addition, the inherent moisture of the product is challenging because probiotics are generally sensitive to water (especially in combination with high temperatures). To date, no technology or skill is believed to deliver adequate protection to probiotics under moderate moisture conditions (i.e., water activity of about 0.2 and higher and up to about 0.4 or higher) and high temperatures during transportation and storage (i.e., temperatures of at least about 30 ℃ and up to and above 40 ℃) when added to nutritional formulas.
In particular, probiotics may provide a variety of benefits to the host, such as maintaining a healthy gastrointestinal flora, enhancing immunity, preventing diarrhea, atopic dermatitis, and other diseases, and the like. Thus, there is a need to administer probiotics in various geographical locations (including tropical climates) where their viability is compromised. Conventional encapsulation and stabilization techniques have chemical compositions that are not suitable for infant formulas and/or use by children, or known techniques have poor stability characteristics that significantly limit commercial opportunities.
There is therefore a need for a stabilizing technique and a stabilized bacterial mixture using acceptable ingredients of infant formula or nutrition for children, which stabilized mixture allows to improve the stability properties so that the probiotic can be transported in a wide variety of geographical locations and climates, while maintaining a useful shelf life. There is also a need for stabilization techniques to protect probiotics, such as Lactobacillus rhamnosus, for use in nutritional compositions such as infant formulas, supplements and children's products. Indeed, the combination of features including improved stability in combination with nutritional factors provides an improved stabilized mixture suitable for prenatal, infant and child nutrition.
Disclosure of Invention
The invention aims to provide a probiotic composition which can lead probiotics to be stably stored. It is another object of the present invention to provide a nutritional composition comprising a protein source and a probiotic composition. It has surprisingly been found that the probiotic composition of the present invention or a nutritional composition comprising the same exhibits excellent biological stability, in particular biological stability enabling stable storage of the probiotic.
To this end, the invention provides in a first aspect a probiotic composition comprising: probiotic bacteria, phospholipids, glycerides, optionally carbohydrates, and optionally binders, the probiotic composition being in the form of a granulate or powder and having a water content of less than 4%.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, comprises 1 x 10 per 1g6-1×1014Probiotics of cfu, e.g. 1X 109-1×1011cfu of probiotic bacteria. The probiotic added to the probiotic composition may be in the form of its commercial source or may be in a culture expanded form which allows the inclusion of a biologically acceptable substrate such as the probiotic present in yoghurt.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, comprises 5-20 mg of phospholipids per 1g, such as 7.5-12.5 mg of phospholipids.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, comprises 5 to 20mg of glycerides, such as 7.5 to 12.5mg of glycerides per 1 g.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, where necessary, may have added thereto carbohydrates which in the present invention generally function as excipients and/or diluents, in particular for rendering the present probiotic composition in a certain volume/weight, e.g. in case the other components are not sufficient to bring the present probiotic composition to the desired volume and/or weight, this may be achieved by adding an appropriate amount of carbohydrate. Generally, such carbohydrates that function as excipients and/or diluents do not have a positive or negative impact on the biological activity of the probiotic. Thus, the amount of carbohydrate added in the probiotic composition according to the invention is illustrated as being added in a balanced amount, for example expressed as "added to the total amount of the formula", "added to the full amount", "added to the balanced amount" … … etc. or the like.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, where necessary, may have added thereto a binding agent which may cause the various materials in the probiotic composition of the present invention to form granules or powders of a larger size than the original size. Typically, the probiotic composition comprises 5-20 mg of binder per 1g, for example 7.5-12.5 mg of binder.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, further comprising Oryzanol (or Oryzanol). In one embodiment, the probiotic composition comprises 0.5-2 mg of oryzanol per 1g, such as 0.75-1.25 mg of oryzanol.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, further comprising zinc glycinate. In one embodiment, the probiotic composition comprises 0.1-0.5 mg of zinc glycinate per 1g, such as 0.1-0.25 mg of zinc glycinate. It has surprisingly been found that when preparing the probiotic composition of the present invention, the simultaneous addition of suitable amounts of both oryzanol and zinc glycinate together with the probiotic bacteria results in a significantly improved survival stability of the probiotic bacteria in the resulting probiotic composition compared to the addition of neither or only one of these.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said probiotic comprises viable microbial cells.
A probiotic composition according to any of the embodiments of the first aspect of the present invention, wherein said viable microbial cells comprise Lactobacillus rhamnosus (Lactobacillus rhamnosus).
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said viable microbial cells comprise Bifidobacterium longum BB536(Bifidobacterium longum BB536), Bifidobacterium longum subsp. infantis 35624(Bifidobacterium longum subsp. infantis 35624), Bifidobacterium animalis subsp. lactis BB-12(Bifidobacterium animalis subsp. lactis BB-12) or any combination thereof.
A probiotic composition according to any of the embodiments of the first aspect of the present invention, wherein said viable microbial cells comprise Lactobacillus rhamnosus (Lactobacillus rhamnosus).
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said viable microbial cells comprise Bifidobacterium longum BB536(Bifidobacterium longum BB536), Bifidobacterium longum subsp. infantis 35624(Bifidobacterium longum subsp. infantis 35624), Bifidobacterium animalis subsp. lactis BB-12(Bifidobacterium animalis subsp. lactis BB-12) or any combination thereof.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, further comprising pectin.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said phospholipid comprises at least one phospholipid. In one embodiment, the at least one phospholipid comprises lecithin.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said glyceride comprises at least one glyceride. In one embodiment, the at least one glyceride comprises a monoglyceride, a diglyceride, a triglyceride, or any combination thereof.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said probiotic composition is encapsulated in a capsule.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, wherein said capsule further comprises an amount of docosahexaenoic acid.
A probiotic composition according to any one of the embodiments of the first aspect of the present invention, wherein said probiotic composition further comprises non-viable probiotic bacteria.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, wherein said non-viable probiotic may be lactobacillus rhamnosus, bifidobacterium longum BB536, bifidobacterium longum subsp.
The probiotic composition according to any of the embodiments of the first aspect of the present invention, wherein said probiotic composition further comprises at least one additional lipid component, i.e. a lipid component distinct from the phospholipids and glycerides of the present invention.
Further, the second aspect of the present invention provides a method for preparing the probiotic composition according to any of the embodiments of the first aspect of the present invention, which comprises the following steps:
(1) uniformly mixing the probiotics, the oryzanol and the zinc glycinate, and then adding the phospholipid and the glyceride for uniform mixing;
(2) adding powdery carbohydrate into the mixture, mixing, adding binder prepared with water, mixing, and making into wet particulate;
(3) drying the wet granules to reduce the water content to below 4%.
In any embodiment of the second aspect of the invention, the drying process for drying the resulting wet granulation to reduce the moisture to less than 4% may be a drying process known in the art, preferably a drying process at a temperature below 45 ℃. For example a fluid bed drying process at a temperature below 45 c, for example a fluid bed drying process at a temperature below 40 c. Such as freezing at a temperature below 40 ℃, quick freezing, freeze drying, ambient air drying, vacuum drying, spray drying, cryogenic drying, and any combination thereof.
Further, a third aspect of the invention provides a nutritional composition comprising:
a probiotic composition according to any of the embodiments of the first aspect of the invention; and
a protein source for providing nutrition to a user.
According to a third aspect of the invention, the probiotic composition comprises from 0.1 to 20%, such as from 0.5 to 10%, such as from 1 to 5% by weight of the total weight of the nutritional composition. Of course, the proportion of probiotic composition in the nutritional composition may also be adjusted, varied within a broader range, depending on the particular circumstances of the individual user.
According to a third aspect of the invention, the nutritional composition may be a pre-formulation of the probiotic composition of the first aspect of the invention with the protein source, for example by compounding the nutritional composition with the protein source immediately after preparation of the probiotic composition of the first aspect of the invention; the nutritional composition may also be a extemporaneous formulation of the probiotic composition of the first aspect of the present invention with the protein source, for example by packaging the probiotic composition of the first aspect of the present invention separately after it has been prepared and then compounding this probiotic composition with the protein source to produce the nutritional composition at the time of its use or at a future time of use, for example immediately before use within 3 months or within 1 month or within 1 week.
The probiotic composition of the first aspect of the invention acts as a material (product, finished product or intermediate) for stabilizing the probiotic bacteria relative to the nutritional composition of the third aspect of the invention, and may also be referred to herein as a protective matrix containing the probiotic bacteria, which contains the probiotic bacteria and a protective component for stabilizing the probiotic bacteria, such as the phospholipids, glycerides, oryzanol, zinc glycinate, and the like, as described herein.
According to a third aspect of the invention, the nutritional composition is a powdered infant formula.
According to a third aspect of the invention, the nutritional composition further comprises at least one additional lipid component, i.e. a lipid component distinct from the phospholipids and glycerides of the invention.
According to a third aspect of the invention, the nutritional composition further comprises non-viable probiotic bacteria.
Any technical feature possessed by any one aspect of the invention or any embodiment of that aspect is equally applicable to any other embodiment or any embodiment of any other aspect, so long as they are not mutually inconsistent, although appropriate modifications to the respective features may be made as necessary when applicable to each other. Various aspects and features of the disclosure are described further below.
All documents cited herein are incorporated by reference in their entirety and to the extent such documents do not conform to the meaning of the present invention, the present invention shall control. Further, the various terms and phrases used herein have the ordinary meaning as is known to those skilled in the art, and even though such terms and phrases are intended to be described or explained in greater detail herein, reference is made to the term and phrase as being inconsistent with the known meaning and meaning as is accorded to such meaning throughout this disclosure.
In some embodiments, the present disclosure relates to a nutritional composition comprising a protein source and a probiotic, wherein the probiotic is stabilized in a protective matrix comprising at least one phospholipid and at least one glyceride.
In these embodiments, the nutritional composition comprises viable microbial cells, such as viable lactobacillus rhamnosus cells. Furthermore, the matrix may comprise a hydrolyzed protein source, pectin, additional lipids, or any combination thereof. The nutritional composition may be a powdered formula, such as a powdered infant formula. Moreover, the at least one glyceride may comprise a monoglyceride, a diglyceride, or any combination thereof.
In another embodiment, the present disclosure relates to a method of protecting viable probiotics of a nutritional composition for a powder, the method comprising the steps of: (i) providing a viable probiotic, (ii) preparing a protective matrix for the probiotic by blending together at least one phospholipid and at least one glyceride, and (iii) combining the viable probiotic, the protective matrix and water to produce a mixture; and (iv) drying the mixture of step (iii) to a final moisture content of about 4% or less. Such a method may comprise the further steps of: a) (iii) adding the dry mixture to a powdered nutritional product, or b) encapsulating the dry mixture of step (iv) in a capsule. In such an embodiment, the viable probiotic may be lactobacillus rhamnosus (which may also be labelled LGG in the present invention), bifidobacterium longum BB536 (which may also be labelled BB536 in the present invention), bifidobacterium longum subsp infantis 35624 (which may also be labelled 35624 in the present invention), bifidobacterium animalis subsp lactis BB-12 (which may also be labelled BB-12 in the present invention), or any combination thereof.
These and other aspects, which will become apparent to the skilled person upon consideration of the following description, may be implemented by providing a mixture of stabilized phospholipids and glycerides and oryzanol and zinc glycinate for biological materials, such as probiotics, to provide improved stability of the biological material, resulting in improved, long-term viability of the biological material. In one embodiment, the stabilized mixture advantageously extends the shelf life of the probiotic. The stabilized mixture may be combined with the probiotic in a variety of ways including freeze drying, air drying, vacuum drying, spray drying, and any combination thereof for preserving the probiotic.
It is to be understood that both the foregoing general description and the following detailed description provide embodiments of the disclosure and are intended to provide an overview or framework for understanding the nature and character of the disclosure as it is claimed.
Reference now will be made in detail to embodiments of the disclosure, one or more examples of which are described below. The various examples are provided by way of explanation of the nutritional compositions of the present disclosure and are not limiting. Indeed, it will be apparent to those skilled in the art that various modifications and variations can be made to the teachings of the disclosure without departing from the scope thereof. For instance, features illustrated or described as part of one embodiment, can be used with another embodiment to yield a still further embodiment.
Thus, it is intended that the present disclosure cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents. Other objects, features and aspects of the present disclosure are disclosed in or are apparent from the following detailed description. It is to be understood by one of ordinary skill in the art that the present discussion is a description of exemplary embodiments only, and is not intended as limiting the broader aspects of the present disclosure.
The present disclosure provides stabilization techniques and stabilization mixtures (also referred to herein as "stabilization mixtures" or "protective matrices") that can be used to improve the stability of biological materials (also referred to herein as "substrates"). In embodiments of the present disclosure, the stabilized substrate may be a probiotic, wherein upon ingestion of the nutritional composition comprising the stabilized probiotic, various health benefits associated with the stabilized probiotic may be imparted to the individual.
Although probiotics are believed to have nutritional benefits, it is generally believed that the beneficial effects of probiotics are maximized if the probiotic microorganisms are ingested by the subject while the microorganisms are still alive. Thus, viable probiotics need to withstand the conditions of manufacture and deployment into consumable nutritional compositions (e.g., food or beverages), as well as subsequent transport and storage times before the product is ingested and introduced into the gastrointestinal tract of a subject. Many conventional probiotic compositions employ extremely high counts of viable cells, and it is understood that a significant number of cells eventually lose viability and die during the production process, transport and storage.
By practicing the present disclosure, a lipid component comprising at least one phospholipid and/or at least one glyceride is added to a protective matrix. The phospholipids and/or glycerides improve the stability of the matrix-protected probiotic. Thus, infant and child compatible probiotics may be stabilized with a matrix comprising at least one phospholipid and/or at least one glyceride. In some embodiments, the matrix further comprises at least one additional lipid, i.e., at least one lipid other than phospholipids and glycerides. Since probiotics exhibit improved viability, the stabilized probiotics may be used in a variety of environments. Advantageously, since the probiotic will maintain viability even after extended transport and storage times (due to the improved stability of the stabilized mixture), the probiotic stabilized by the protective matrix of the present disclosure may be added to a nutritional composition and transported over extended distances subject to potentially damaging temperature fluctuations.
The method of the invention for providing stabilization to a probiotic may comprise the use of a matrix for stabilizing a biological material, wherein the matrix comprises phospholipids, glycerides, oryzanol, zinc glycinate and further components, such as one or more further lipids, one or more carbohydrates and/or a compound binder.
Although the protective matrix may be used for a wide variety of substances, in one embodiment it is used to protect at least one probiotic, such as lactobacillus rhamnosus. It will be appreciated that rhamnose lactic acid bacteria have a relatively good biostability, while having a high affinity for human intestinal mucosal cells. When used as probiotics, rhamnose lactic acid bacteria are believed to colonize the gut and balance the intestinal microbiota.
The protective matrix provides improved stability of the probiotic bacteria, which means that a larger percentage of the probiotic cells survive after processing, transport and storage conditions. In particular, the shelf life of viable probiotics is improved when compared to other known stabilization techniques.
The protective matrix of the present disclosure can be used in a variety of methods of forming stabilized probiotic products. These methods include freezing, quick freezing, freeze drying, ambient air drying, vacuum drying, spray drying, cryogenic drying, and any combination thereof. The resulting stabilized probiotics, whether alone or integrated into nutritional compositions, have effective viability over a wide range of temperatures and conditions while exhibiting improved shelf life. Furthermore, the stabilized probiotic may be added to a wide variety of prenatal, infant and children's nutritional products to improve their intestinal microbiota while providing nutrition to the infant or child.
Thus, in one embodiment, the present disclosure relates to a method for stabilizing a biological material in a nutritional composition. Another embodiment is a protective matrix for probiotics. Yet another embodiment is a method of increasing the shelf life of a probiotic, the method comprising stabilizing the probiotic with a stabilizing mixture comprising a phospholipid, a glyceride, oryzanol, zinc glycinate, or any combination thereof.
The present disclosure provides novel stabilized mixtures and methods of providing stability and protection to biological materials, such as viable microorganisms. The present disclosure includes a stabilized mixture comprising phospholipids and glycerides that together provide a protective matrix, which results in an improved shelf life relative to unprotected probiotics.
In the present invention, the terms "protective matrix", "stabilizing matrix" and "stabilizing mixture" are used interchangeably throughout the disclosure of the present invention.
An "effective amount" as used herein is generally defined as the amount of an agent that provides an observable result in a subject to which it is administered.
By "nutritional composition" is meant a substance or formulation that meets at least a portion of the nutritional needs of a subject. The terms "nutritional agent", "nutritional formula", "enteral nutritional agent" and "nutritional supplement" are used throughout the present disclosure as non-limiting examples of nutritional compositions. Furthermore, "nutritional composition" may refer to a ready-to-use form of a liquid, powder, gel, paste, solid, concentrate, suspension or enteral formula, oral formula, infant formula, pediatric subject formula, pediatric formula, growing-up milk and/or adult formula.
The term "enteral" means deliverable through or within the gastrointestinal or digestive tract. "enteral administration" includes oral feeding into the digestive tract, intragastric feeding, administration through the pylorus, or any other administration. "administration" is more general than "enteral administration" and includes parenteral administration or any other route of administration whereby a substance is taken into the body of a subject.
By "pediatric subject" is meant a person less than 13 years of age. In some embodiments, a pediatric subject refers to a human subject that is less than 8 years old. In other embodiments, a pediatric subject refers to a human subject with an age of 1-6 years. In yet another embodiment, a pediatric subject refers to a human subject with an age of 6-12 years.
"infant" means a human subject ranging in age from birth to no more than 1 year of age and includes infants aged 0-12 months corrected. The phrase "corrected age" means the chronological age of the infant minus the amount of time the infant was prematurely delivered. Thus, if the gestation has reached term, the corrected age is that of the infant. The term infant includes low birth weight infants, very low birth weight infants and premature infants. A "preterm infant" is an infant born after less than about 37 weeks of gestation. As used herein, "term infant" means an infant born after at least about 37 weeks of gestation.
By "child" is meant a subject with an age in the range of 12 months to about 12 years. In some embodiments, the child is a subject between 1 and 12 years of age. In other embodiments, the term "child" refers to a subject from 1 to about 6 years of age, or from about 7 to about 12 years of age. In other embodiments, the term "child" refers to any range of ages from 12 months to about 12 years.
"nutritional product for children" refers to a composition that meets at least a portion of the nutritional needs of a child. Growing Up Milks (GUM) are examples of nutritional products for children.
As used herein, "hydrolyzed protein" refers to a protein hydrolysate. Within the present disclosure, hydrolyzed protein and protein hydrolysate are used interchangeably to describe protein hydrolysates; fully hydrolyzed protein is used to describe a protein hydrolysate wherein at least 70%, more preferably at least about 90% of the hydrolyzed protein has a molecular weight of less than 2000 daltons.
The term "degree of hydrolysis" refers to the extent to which peptide bonds are destroyed by the hydrolysis process.
The term "protein-free" means containing an immeasurable amount of protein, as measured by standard protein detection methods, such as sodium dodecyl (lauryl) sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) or size exclusion chromatography. In some embodiments, the nutritional composition is substantially free of protein, wherein "substantially free" is defined below.
By "infant formula" is meant a composition that meets at least a portion of the nutritional needs of an infant.
The term "growing-up milk" refers to a broad category of nutritional compositions intended for use as part of a diverse diet in order to support the normal growth and development of children aged from about 1 to about 6 years.
"milk-based" means comprising at least one component drawn or extracted from the mammary gland of a mammal. In some embodiments, the milk-based nutritional composition comprises components of milk derived from domesticated ungulates, ruminants or other mammals, or any combination thereof. Also, in some embodiments, the milk-based representation comprises bovine casein, whey, lactose, or any combination thereof. Also, "milk-based nutritional composition" may refer to any composition comprising any milk-derived or milk-based product known in the art.
By "nutritionally complete" is meant a composition that can be used as the sole source of nutrition that will supply substantially all of the daily quantities of vitamins, minerals, and/or trace elements required, as well as protein, carbohydrate, and lipids. Indeed, "nutritionally complete" describes a nutritional composition that provides sufficient amounts of carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditionally essential amino acids, vitamins, minerals, and energy required to support normal growth and development in a subject.
A composition that is "nutritionally complete" for a preterm infant will, by definition, qualitatively and quantitatively provide all the carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditionally essential amino acids, vitamins, minerals and energy in sufficient quantities for the preterm infant to grow. A composition that is "nutritionally complete" for a term infant will, by definition, qualitatively and quantitatively provide all carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditionally essential amino acids, vitamins, minerals and energy in sufficient amounts for growth of the term infant. A composition that is "nutritionally complete" for a child will, by definition, qualitatively and quantitatively provide all carbohydrates, lipids, essential fatty acids, proteins, essential amino acids, conditionally essential amino acids, vitamins, minerals, and energy in sufficient quantities for growth of the child.
The term "essential" when applied to nutrients refers to any nutrient that is not synthesized by the body in sufficient quantities to grow and maintain health normally and therefore must be supplied by the diet. The term "conditionally essential" when applied to nutrients means that nutrients must be supplied by the diet under conditions where the body does not have access to sufficient amounts of precursor compounds for endogenous synthesis.
The term "probiotic" means a microorganism with low or no pathogenicity that exerts beneficial effects on the health of the host. By "viable probiotic" is meant a viable or active microorganism that exerts a beneficial effect on the health of the host.
The term "inactivated probiotic" or "inactivated LGG" denotes a probiotic wherein the metabolic activity or reproductive capacity of the probiotic or rhamnose lactic acid bacteria gg (LGG) organism is reduced or impaired. In one embodiment, the probiotic may be viable or non-viable. The term "viable" as used herein refers to a viable microorganism. The term "non-viable" or "non-viable probiotic" denotes non-viable probiotic micro-organisms, cellular components thereof and/or metabolites thereof. Such non-viable probiotics may be heat killed or otherwise inactivated, but they retain the ability to beneficially affect the health of the host. Probiotics useful in the present disclosure may be naturally occurring, synthetic, or developed by genetic manipulation of organisms, whether such sources are now known or later developed.
By "prebiotic" is meant a limited amount of bacteria in the digestive tract that beneficially affects the host's indigestible food ingredients or may improve the host's health by selectively stimulating the growth and/or activity of the individual.
"phytonutrients" means chemical compounds that occur naturally in plants. The phytonutrients may be contained in any plant-derived material or extract. The term "phytonutrients" encompasses a wide variety of broad classes of compounds produced by plants, such as polyphenolic (polyphenolic) compounds, anthocyanins, proanthocyanidins and flavan-3-ols (i.e. catechins, epicatechins), and may for example be derived from fruit, seed or tea extracts. Furthermore, the term phytonutrients includes all carotenoids, phytosterols, thiols and other plant derived compounds.
"beta-glucan" means all beta-glucans, including certain types of beta-glucans, such as beta-1, 3-glucan or beta-1, 3; 1, 6-glucan. And, β -1, 3; 1, 6-glucan is a class of beta-1, 3-glucans. Thus, the term "β -1, 3-glucan" includes β -1, 3; 1, 6-glucan.
"pectin" means any naturally occurring oligo-or polysaccharide comprising galacturonic acid that can be found in the cell wall of a plant. Furthermore, pectin may comprise molecules having various chain lengths formed by a D-galactopyranosyl group. Different kinds and grades of pectins with varying physical and chemical properties are known in the art. Indeed, the structure of pectin can vary significantly between plants, between tissues, and even between single cell walls. Generally, pectin consists of negatively charged acidic sugars (galacturonic acid) and some of the acidic groups are in the form of methyl ester groups. The degree of esterification of pectin is a measure of the percentage of carboxyl groups attached to the galactopyranosyluronic acid unit esterified with methanol.
Pectins with a degree of esterification of less than 50% (i.e. less than 50% of the carboxyl groups are methylated to form methyl ester groups) are classified as low-ester, low-methoxy or low-methylated ("LM") pectins, while those with a degree of esterification of 50% or more than 50% (i.e. more than 50% of the carboxyl groups are methylated) are classified as high-ester, high-methoxy or high-methylated ("HM") pectins. Very low ("VL") pectins, a subset of low methylated pectins, have a degree of esterification of less than about 15%. Moreover, in some embodiments, the pectin molecules have the ability to interact with themselves and/or with divalent cations such as calcium by electrostatic interactions to form high molecular weight networks and gels. The degree of esterification affects the degree of these intermolecular interactions.
"lipid" refers to a hydrophobic molecule that may include free fatty acids and/or molecules in which the fatty acids are esterified to the hydroxyl groups of: (i) a sugar, such as a mono-or disaccharide, (ii) an alcohol, wherein the alcohol may comprise, for example, a long or medium carbon chain length (10-20 units), or (iii) glycerol. These esterified or free fatty acids may be linear, saturated or unsaturated. Also, in some embodiments, the lipid component comprises glycerides, phospholipids, sphingosine-derived molecules (e.g., gangliosides, cerebrosides, and sphingomyelins), or any combination thereof. And in certain embodiments, the lipid component further comprises molecules of a hydrophobic nature, such as steroids, carotenoids and/or fat-soluble vitamins.
"glyceride" means any molecule derived from a glycerol or sugar backbone in which one or more of the hydroxyl groups are esterified with a fatty acid. In some embodiments, the glyceride component comprises monoglycerides, diglycerides, monosaccharide esters, disaccharide esters, or a combination thereof.
"phospholipid" means any molecule derived from phosphatidic acid, wherein one or two carbons from the glycerol backbone are esterified to a fatty acid and one of the capped carbons is esterified to a phosphate group. In certain embodiments, a phosphate group is a phosphodiester that is also esterified to another functional molecule, such as choline, serine, or inositol. In some embodiments, the phospholipid component comprises glycerophospholipids and/or phosphorylglycerols. Furthermore, the phospholipid component may comprise a lysophospholipid in which one of the carbons from the glycerol backbone remains an unesterified hydroxyl group. And in some embodiments, the phospholipid component comprises lecithin, which may be extracted, for example, from soy or from egg yolk and may comprise a combination of phosphatidyl-choline, phosphatidyl-inositol, and phosphatidyl serine. In some embodiments, the phospholipid may comprise soy lecithin, egg lecithin, milk fat-derived phospholipids, or a combination thereof.
All percentages, parts and ratios used herein are by weight of the total nutritional composition including the stabilized probiotic, unless otherwise specified.
All amounts specified for "daily" administration may be given in one unit dose, a single delivery, or two or more doses or servings given over the course of a 24 hour period.
The nutritional compositions of the present disclosure may be substantially free of any optional or selected ingredients described herein, provided that the remaining nutritional composition still comprises all of the required ingredients or features described herein. In this context, and unless otherwise specified, the term "substantially free" means that the selected composition may contain less than a functional amount of optional ingredients, typically less than 0.1% by weight, and also contain 0% by weight of such optional or selected ingredients.
All references to singular features or limitations of the present disclosure shall include the corresponding plural features or limitations and vice versa unless otherwise specified or clearly contradicted by context of reference.
All combinations of methods or method steps used herein can be performed in any order, unless otherwise specified or clearly contradicted by context in which the combination is referred to.
The methods and compositions of the present disclosure (including components thereof) can comprise, consist of, or consist essentially of the essential elements and limitations of the embodiments described herein, as well as any additional or optional ingredients, components, or limitations described herein or otherwise useful in nutritional compositions.
The term "about" as used herein should be considered to refer to two numbers that specify the end points of any range. Any reference to a range should be considered to provide support for any subset of the ranges.
In the practice of the present disclosure, at least one phospholipid, at least one glyceride, oryzanol, zinc glycinate are used as components of the protective matrix for stabilizing the biological material.
The use of phospholipids and/or glycerides in the protective matrix of the present disclosure provides excellent protection for probiotics, including lactobacillus rhamnosus, over what is previously known in the art.
The lipid component of the protective matrix may include, but is not limited to, animal sources such as milk fat, butter fat, milk phospholipids, egg yolk lipids, egg yolk phospholipids; marine sources, such as fish oil, marine oil, single cell oil; vegetable and vegetable oils, such as corn oil, rapeseed oil, sunflower oil, soybean phospholipids, palm olein, coconut oil, high oleic sunflower oil, evening primrose oil, canola oil, olive oil, linseed (linseed) oil, cottonseed oil, high oleic safflower oil, palm stearin, palm kernel oil, wheat germ oil; medium chain triglyceride oils and emulsions and esters of fatty acids; and any combination thereof.
In the present invention, the probiotic composition may also be referred to as a "protective matrix" comprising the probiotic bacteria or may also be referred to as a "stabilized mixture" or may also be referred to as a "stabilized probiotic" or other similar terms. In one embodiment, the "stabilized mixture" comprises from about 0.5 to about 50 grams of lipid component per 100 grams of mixture on a dry basis. In certain embodiments, the stabilized mixture comprises from about 2.5 to about 30 grams of the lipid component per 100 grams of the mixture on a dry basis.
The stabilized mixture may comprise a phospholipid component. In certain embodiments, the stabilized mixture comprises, on a dry basis, from about 0.5 to about 2 grams of the phospholipid component per 100 grams of the mixture. In some embodiments, the stabilized mixture comprises, on a dry basis, from about 0.75 to about 1.25 grams of phospholipid component per 100 grams of mixture.
Likewise, the stabilized mixture may comprise a glyceride component. In some embodiments, the stabilized mixture comprises from about 0.5 to about 2 grams of the glyceride component per 100 grams of the mixture, on a dry basis. In certain embodiments, the stabilized mixture comprises from about 0.75 to about 1.25 grams of the glyceride component per 100 grams of the mixture, on a dry basis. Moreover, the glyceride component can comprise monoglycerides, diglycerides, or any combination thereof.
In certain embodiments, the major component of the stabilized mixture on a dry weight basis is one or more carbohydrates, which may include polysaccharides, disaccharides, and monosaccharides. Indeed, the protective matrix may comprise lactulose, lactosucrose, raffinose, gluco-oligosaccharides, trehalose, inulin, polydextrose, galacto-oligosaccharides, fructo-oligosaccharides, isomalto-oligosaccharides, soy oligosaccharides, lactosucrose, xylo-oligosaccharides, chito-oligosaccharides, manno-oligosaccharides, arabino-oligosaccharides, sialyl-oligosaccharides, fuco-oligosaccharides, gentio-oligosaccharides and/or any combination thereof. In some embodiments, the protective matrix comprises a first carbohydrate selected from the group consisting of: sucrose, maltose, lactose, trehalose, maltotriose, maltodextrin, and any combination thereof having a dextrose equivalent of about 2 to about 6. In certain embodiments, the protective matrix comprises a second carbohydrate selected from the group consisting of: inulin having a dextrose equivalent of greater than about 8, polydextrose, galacto-oligosaccharide, fructo-oligosaccharide, starch, maltodextrin, and any combination thereof.
In some embodiments, the stabilized mixture may comprise, on a dry basis, from about 50 to about 80 grams of the first carbohydrate per 100 grams of the mixture; from about 60 to about 70 grams of the first carbohydrate per 100 grams of the mixture on a dry basis; or from about 65 to about 70 grams of the first carbohydrate per 100 grams of the mixture on a dry basis. The first carbohydrate may be selected from: sucrose, maltose, lactose, trehalose, maltotriose, maltodextrin, and any combination thereof having a dextrose equivalent of about 2 to about 6.
The stabilized mixture may further comprise, on a dry basis, from about 1 to about 10 grams of a second carbohydrate per 100 grams of the mixture; from about 4 to about 6 grams of the second carbohydrate per 100 grams of the mixture on a dry basis; or about 5 grams of the second carbohydrate per 100 grams of the mixture on a dry basis. In some embodiments, the second carbohydrate is selected from: inulin having a dextrose equivalent of greater than about 8, polydextrose, galacto-oligosaccharide, fructo-oligosaccharide, starch, maltodextrin, and any combination thereof.
The other component of the stabilized mixture may be a compound binder (also referred to as a gelling agent) which may act as a thickener and create a gelatinous consistency. The compound binders that may be included in the protective matrices of the present disclosure include alginates such as sodium alginate, pectin, chitosan, carboxymethyl cellulose, mixtures thereof, and the like. The addition of the compound binder provides for the formation of a viscous consistency, which provides for the formation of an effective matrix and structural qualities suitable for subsequent drying.
In some embodiments, the compound binder may form a gel-like material and increase the viscosity of the mixture to which the gel-like material is added. In addition, the compound binder may also make the components easier to mix together. For example, sodium alginate may also have an emulsifier character.
In some embodiments, the stabilized mixture may comprise LM pectin, HM pectin, VL pectin, or any mixture thereof. The contained pectin can be dissolved in water.
The pectins used herein typically have a peak molecular weight of 8,000 daltons or greater. The pectin of the present disclosure has a preferred peak molecular weight of 8,000 to about 500,000, more preferably about 10,000 to about 200,000 and most preferably about 15,000 to about 100,000 daltons. In some embodiments, the pectin of the present disclosure may be a hydrolyzed pectin. In certain embodiments, the protective matrix comprises a hydrolyzed pectin having a molecular weight less than the molecular weight of an intact or unmodified pectin. The hydrolyzed pectin of the present disclosure can be prepared by any means known in the art to reduce molecular weight. Examples of such means are chemical hydrolysis, enzymatic hydrolysis and mechanical shearing. The preferred way to reduce the molecular weight is by alkaline or neutral hydrolysis at elevated temperature. In some embodiments, the protective matrix comprises a partially hydrolyzed pectin. In certain embodiments, the partially hydrolyzed pectin has a molecular weight less than that of intact or unmodified pectin, but greater than 3,300 daltons.
The stabilized mixture may comprise from about 0.5 to about 5 grams of compound binder (e.g., sodium alginate and/or pectin) on a dry basis per 100 grams of the mixture. In certain embodiments, the stabilized mixture comprises, on a dry basis, from about 1 to about 3 grams of compound binder per 100 grams of the mixture. And in one embodiment, the stabilized mixture comprises about 2 grams of compound binder per 100 grams of mixture on a dry basis.
Furthermore, the stabilized mixture may further comprise at least one starch, a source of starch and/or a starch component. In some embodiments, the stabilized mixture may, for example, comprise native or modified starches, such as waxy corn starch, waxy rice starch, waxy potato starch, waxy tapioca starch, corn starch, rice starch, potato starch, tapioca starch, wheat starch, or any mixture thereof.
The stabilized mixture may also include additional ingredients that provide other benefits to the probiotic or the individual ingesting the stabilized probiotic. These ingredients may include minerals, vitamins, antioxidants, trace elements, sterols, antioxidants, fatty acids, functional molecules, and any combination thereof. Other ingredients may include resistant starch, high amylose starch, guar gum and locust bean gum, agar, xanthan gum, carrageenan, dextran, and any combination thereof.
In some embodiments, the stabilized mixture may comprise from about 5 to about 90 grams of probiotics and/or other biological materials per 100 grams of the mixture on a dry basis. In some embodiments, the stabilized mixture comprises from about 50 to about 90 grams of probiotic and/or other biological material per 100 grams of stabilized mixture. In certain embodiments, the stabilized mixture comprises from about 9 to about 12 grams of probiotics and/or other biological materials per 100 grams of the mixture on a dry basis. And in one embodiment, the stabilized mixture comprises from about 10.2 to about 11.4 grams of probiotic and/or other biological material per 100 grams of mixture on a dry basis. In another embodiment, the concentration of probiotic bacteria, such as LGG, in the protective matrix is about 1 x 106About 1X 1014cfu/g protective matrix, more preferably about 1X 109About 1X 1011cfu/g protective matrix.
The stabilized mixture may be used to provide stability to probiotic organisms that may exert beneficial effects on the health and welfare of an individual. Examples of suitable probiotics include, but are not limited to, yeasts (e.g. saccharomyces cerevisiae), moulds (e.g. Aspergillus, Rhizopus, Mucor) and bacteria (e.g. lactobacillus). Specific examples of suitable probiotic microorganisms are: aspergillus niger (Aspergillus niger), Aspergillus oryzae (A.oryzae), Bacillus coagulans (Bacillus coaguluns), Bacillus lentus (B.lentus), Bacillus licheniformis (B.licheniformis), Bacillus mesentericus (B.mesentericus), Bacillus pumilus (B.pumilus), Bacillus subtilis (B.subtilis), Bacillus natto (B.natto), Bifidobacterium adolescentis (Bifidobacterium adolescentis), Bifidobacterium animalis (B.animalis), Bifidobacterium breve (B.breve), Bifidobacterium bifidum (B.bifidum), Bifidobacterium infantis (B.infantis), Bifidobacterium lactis (B.lactis), Bifidobacterium longum (B.longum), Bifidobacterium longum BB536(B.longum BB536), Bifidobacterium longum AH1206(NCIMB 171 41382), Bifidobacterium longum 1206(NCIMB 10135387), Bifidobacterium longum BB 35387), Bifidobacterium longum 35387 (NCIMB 35387), Bifidobacterium longum (NCIMB 35624), Bifidobacterium longum 351205), Bifidobacterium longum (NCIMB 35387), Bifidobacterium longum) and Bifidobacterium longum (NCIMB 35387), Bifidobacterium longum 35624), Bifidobacterium longum (NCIMB 35387) 3, Bifidobacterium longum) Bifidobacterium pseudolongum (b. pseudolongum), bifidobacterium thermophilum (b. thermophilum), Candida pintolepsilon, Clostridium butyricum (Clostridium butyricum), Enterococcus cremoris (Enterococcus cremoris), Enterococcus dibutylosus (e.diacetylactis), Enterococcus faecalis (e.faecium), Enterococcus intermedius (e.intermedius), Enterococcus lactis (e.lactis), e.mutdi, Enterococcus thermophilus (e.thermophilus), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus digestus (l.alimentarius), Lactobacillus amylovorus (l.amylovorus), Lactobacillus crispatus (l.crispatus), Lactobacillus brevis (l.faecalis), Lactobacillus plantarum (l.souvenius), Lactobacillus fibrous (l.lactis), Lactobacillus lactis (l.lactis), Lactobacillus plantarum (l.l.l.lactis), Lactobacillus plantarum), Lactobacillus casei (l.lactis), Lactobacillus lactis (l.l.r), Lactobacillus lactis (l.l.l.r), Lactobacillus lactis (l.l.r) Lactobacillus rhamnosus, lactobacillus rhamnosus GG (ATCC No. 53103, which may also be referred to herein as LGG), lactobacillus sake (l.sakei), and lactobacillus salivarius (l.salivarius), and any combination thereof. In one embodiment, the stabilized probiotic may be viable or non-viable. Stabilized probiotics useful in the present disclosure may be naturally occurring, synthetic, or developed by genetic manipulation of organisms, whether such new sources are now known or later developed.
In one embodiment of the present disclosure, rhamnose lactic acid bacteria GG are used as probiotics that can be stabilized by the protective matrix of the present disclosure. Lactobacillus rhamnosus GG is described in U.S. patent application 4,839,281 to Sharwood et al, which is incorporated herein by reference in its entirety. In particular, Sharwood et al describe lactobacillus rhamnosus GG as a species in which the bacteria have significant adhesion to the cells in the intestine while being able to survive at low pH and produce large amounts of lactic acid.
The selected probiotic bacteria are preferably concentrated to a wet pasty consistency prior to combining with the stabilized mixture components of the present disclosure. Starting with probiotics in dry form is also an alternative. The concentration levels of the probiotics selected include concentrations of about 3X to about 20X, although lesser or greater concentrations may be included, depending on the particular probiotic biomass and subsequent processing steps.
In general, the preparation of stabilized probiotics may generally comprise the following steps: concentrating the selected probiotic or probiotics; providing components of the stabilized mixture in the required amounts; mixing the stabilized mixture component with concentrated probiotics; drying the stabilized probiotic. The stabilized probiotic is optionally packaged or combined with other components, e.g. into a nutritional composition of the third aspect of the invention or referred to as a nutritional product, e.g. an infant formula.
In some embodiments, the present disclosure relates to methods of protecting viable probiotics for use in nutritional compositions, which may include the steps of: providing viable probiotic bacteria, preparing a protective matrix for the probiotic bacteria by blending: phospholipids, glycerides and any other desired combination, followed by combining the viable probiotic, the protective matrix and water to produce a mixture and drying the mixture to a final moisture content of about 4% or less, and further adding the dried mixture to a powder comprising a protein source to make a nutritional product, or encapsulating the dried mixture for temporary use by a user.
In optimizing the stabilization of the probiotic, various ingredients may be varied in some embodiments as described herein and as shown in formulas a-D. In some embodiments, the stabilized mixture may comprise about 10% -50% (w/w) lipid. In some embodiments, the stabilized mixture may comprise from about 5% to about 25% (w/w) of the phospholipid component and from about 5% to about 25% (w/w) of the glyceride component. In other embodiments, the stabilized mixture may comprise from about 5% to about 25% (w/w) phospholipid component, from about 2.5 to about 12.5% (w/w) glyceride component, and from about 2.5% to about 12.5% (w/w) additional/other lipid component. In yet another embodiment, the total lipids of the stabilized mixture may comprise about 3% to about 17% (w/w) of the phospholipid component, about 3 to about 17% (w/w) of the glyceride component, and about 3% to about 17% (w/w) of the additional/other lipid component.
The stabilized probiotic may be packaged and marketed or may be additionally combined with a wide variety of nutritional products or encapsulated with other functional components such as docosahexaenoic acid. Such nutritional products may include both infant formulas and children's products for applications in which it is desirable to add probiotics to nutritional products that necessitate improved shelf life and stability.
Formula a sample embodiments of stabilized probiotic mixture/protective matrix according to the present disclosure are presented: probiotic bacteria (e.g., LGG, BB536, AH1206, BB-12 or 35624, etc.): 50-90 g/100g (wet basis), phospholipid: 5-25 g/100g (wet basis), glyceride: 5-25 g/100g (wet basis).
Formulation B presents sample embodiments of the stabilized probiotic mixture/protective matrix according to the present disclosure: probiotic bacteria (e.g., LGG, BB536, AH1206, BB-12 or 35624, etc.): 50-90 g/100g (dry basis), total lipids: 10-50 g/100g (dry basis), phospholipids: 5-25 g/100g (dry basis), glyceride: optionally, other lipids: 5-25 g/100g (dry basis).
Formulation C presents yet another example embodiment of a stabilized probiotic mixture/protective matrix according to the present disclosure: probiotic bacteria (e.g., LGG, BB536, AH1206, BB-12 or 35624, etc.): 50-90 g/100g (dry basis), total lipids: 10-50 g/100g (dry basis), phospholipids: 5-25 g/100g (dry basis), glyceride: 2.5-12.5 g/100g (dry basis), other lipids: 2.5-12.5 g/100g (dry basis).
Formula D also provides example embodiments of the stabilized probiotic mixture/protective matrix according to the present disclosure: probiotic bacteria (e.g., LGG, BB536, AH1206, BB-12 or 35624, etc.): 90-50 g/100g (dry basis), total lipids: 10-50 g/100g (dry basis), phospholipids: 3.33-16.7 g/100g (dry basis), glycerides: 3.33-16.7 g/100g (dry basis), other lipids: 3.33-16.7 g/100g (dry basis).
Further, as shown in the above formulas a-D, the protective matrix, i.e. the probiotic composition, is in dry matter (in the form of granules or powder) with a water content of less than 4%. In one embodiment, the probiotic composition in dry matter comprises 1 x 10 per 1g thereof6-1×1014Probiotics of cfu, e.g. 1X 109-1×1011cfu of probiotic bacteria. In one embodiment the probiotic composition in dry matter comprises 5-20 mg of phospholipids per 1g, such as 7.5-12.5 mg of phospholipids. In one embodiment the probiotic composition in dry matter comprises 5-20 mg of glycerides per 1g, such as 7.5-12.5 mg of glycerides. Further, in one embodiment, oryzanol is also comprised in the probiotic composition in dry matter form. In one embodiment the probiotic composition in dry matter comprises 0.5-2 mg of oryzanol per 1g, such as 0.75-1.25 mg of oryzanol. In one embodiment, the probiotic composition in dry matter form further comprises zinc glycinate. In one embodiment the probiotic composition in dry matter comprises 0.1 to 0.5mg of zinc glycinate per 1g, for example 0.1 to 0.25mg of zinc glycinate. Further, in one embodiment, the probiotic composition in dry matter form further comprises a binding agent. In one embodiment the probiotic composition in dry matter comprises 5-20 mg of binder per 1g, for example 7.5-12.5 mg of binder. In one embodiment, the probiotic composition in dry matter form further comprises a carbohydrate. In one embodiment, the probiotic composition in dry matter is added in a balanced amount of carbohydrate per 1 g.
Nutritional products for combination with stabilized probiotics
The stabilized probiotics prepared as described above may be combined with nutritional products to form novel nutritional compositions.
For example, the stabilized probiotic may be combined with a nutritional product, such as an infant formula or a nutritional product for a child, to form a stabilized nutritional composition. In another embodiment, the stabilized probiotic may be combined with a human milk fortifier that is added to human milk in order to enhance the nutritional value of human milk.
Moreover, the stabilized probiotics of the present disclosure may be combined with nutritional products that provide minimal, partial, or complete nutritional support. The nutritional product may be a nutritional supplement or a meal replacement. Indeed, the stabilized probiotic may be intermixed with the food or other nutritional product prior to ingestion by the subject.
The nutritional product for combination with the stabilized probiotic may, but need not, be nutritionally complete. Likewise, the combination of the stabilized probiotic with the nutritional product may result in a nutritionally complete nutritional composition. In one embodiment, the nutritional compositions of the present disclosure are nutritionally complete and comprise suitable types and amounts of lipids, carbohydrates, proteins, vitamins, and minerals.
The stabilized probiotics formed by the present disclosure may be combined with nutritional products provided in any form known in the art, including powders, gels, suspensions, pastes, solids, liquids, liquid concentrates, or ready-to-use products. In one combination, the nutritional product is an infant formula, particularly an infant formula suitable for use as a sole source of nutrition for an infant.
Nutritional products for combination with stabilized probiotics are described that can be administered enterally.
Nutritional compositions comprising stabilized probiotics
Furthermore, the stabilized/protected probiotics prepared as described above may be combined with nutritional products to form novel nutritional compositions.
The nutritional composition may comprise any additional fat or lipid source known or used in the art, including but not limited to animal sources, such as milk fat, butter fat, egg yolk lipids; marine sources, such as fish oil, marine oil, single cell oil; vegetable and vegetable oils, such as corn oil, rapeseed oil, sunflower oil, soybean oil, palm olein, coconut oil, high oleic sunflower oil, evening primrose oil, rapeseed oil, olive oil, linseed (linseed) oil, cottonseed oil, high oleic safflower oil, palm stearin, palm kernel oil, wheat germ oil; medium chain triglyceride oils and emulsions and esters of fatty acids; and any combination thereof. The amount of lipid or fat in the nutritional composition typically varies from about 1 to about 7g/100 kcal.
Furthermore, the nutritional composition may comprise a source of milk protein. Sources of milk protein may include, but are not limited to, milk protein powder, milk protein concentrate, milk protein isolate, skim milk solids, skim milk, defatted dried milk, whey protein isolate, whey protein concentrate, sweet whey, acid whey, casein, acid casein, caseinate (e.g., sodium caseinate, sodium calcium caseinate, calcium caseinate), and any combination thereof.
In certain embodiments, the nutritional composition may comprise intact proteins. In other embodiments, the protein of the nutritional composition is provided as a combination of both intact protein and partially hydrolyzed protein having a degree of hydrolysis of about 4% to 10%. Since both the stabilizer and the protein of the nutritional product comprise hydrolyzed protein only, certain of these embodiments may be extremely hypoallergenic. In yet another embodiment, the nutritional composition may be supplemented with glutamine-containing peptides.
Whey of the protein source of the nutritional composition: the casein ratio may be similar to that found in human breast milk. In one embodiment, the protein source of the nutritional composition comprises from about 40% to about 80% whey protein. In another embodiment, the protein source may comprise about 20% to about 60% casein. The amount of protein in the nutritional composition typically varies from about 1 to about 7g/100 kcal.
In other embodiments, the nutritional composition comprises lactoferrin that maintains its stability and activity in the human intestinal tract against certain undesirable bacterial pathogens.
In some embodiments, the nutritional compositions described herein may further comprise non-human lactoferrin, non-human lactoferrin produced by a genetically modified organism, and/or human lactoferrin produced by a genetically modified organism. Lactoferrin is generally described as an 80 kilodalton glycoprotein with the structure of two nearly identical leaves (lobes) that include an iron binding site. Lactoferrin has been recognized to have bactericidal and antimicrobial activity. In at least one embodiment, the lactoferrin is bovine lactoferrin.
Unexpectedly, the forms of lactoferrin included herein retain relevant activity even when exposed to low pH (i.e., below about 7 and even as low as about 4.6 or less) and/or high temperature (i.e., above about 65 ℃ and up to about 120 ℃) conditions that are expected to disrupt or severely limit the stability or activity of human lactoferrin or recombinant human lactoferrin. These low pH and/or high temperature conditions may be expected during certain processing regimes (e.g., heat sterilization processes) for nutritional compositions of the types described herein.
In one embodiment, lactoferrin is present in the nutritional composition in an amount of about 5mg/100kcal to about 16mg/100 kcal. In another embodiment, lactoferrin is present in an amount of about 9mg/100kcal to about 14mg/100 kcal. In yet another embodiment, the nutritional composition may comprise from about 2mg to about 200mg lactoferrin per 100 kcal. And in certain embodiments, the nutritional composition may comprise from about 90mg to about 148mg lactoferrin per 100 kcal.
The nutritional composition may further comprise TGF- β. In some embodiments, the level of TGF- β may be from about 0.0150(pg/μ g) ppm to about 0.1000(pg/μ g) ppm. In other embodiments, the level of TGF- β in the final composition comprising the stabilized probiotic is from about 0.0225(pg/μ g) ppm to about 0.0750(pg/μ g) ppm.
In some embodiments of the nutritional compositions, the level of TGF- β is from about 2500pg/mL to about 10,000pg/mL, more preferably from about 3000pg/mL to about 8000 pg/mL. In one embodiment, the ratio of TGF- β 1 to TGF- β 2 is in the range of about 1:1 to about 1:20, or more specifically, in the range of about 1:5 to about 1: 15.
In some embodiments, the bioactivity of TGF- β in the nutritional composition is enhanced by the addition of a bioactive whey fraction. Any raw material known in the artA fraction of whey that is active may be used in these embodiments, provided that the desired results are obtained. In one embodiment, such a bioactive whey fraction may be a whey protein concentrate. In a particular embodiment, the whey protein concentrate may be800 from Glanbia Nutritions.
The nutritional composition may comprise an amount of probiotic bacteria in addition to the stabilized probiotic bacteria. When the stabilized probiotic is combined with a nutritional product, the resulting nutritional composition may comprise effective to provide about 1 x 10 to the subject4About 1X 1010Colony forming units (cfu)/kg body weight/total amount of probiotic bacteria per day. In other embodiments, the amount of probiotic may be about 1 × 106About 1X 109change in cfu/kg body weight/day. In even other embodiments, the nutritional composition may comprise a nutritional composition effective to provide about 1 x 106Probiotic bacteria in an amount of cfu/kg body weight/day.
In certain embodiments, the nutritional compositions of the present disclosure comprise about 1 x 106Probiotic bacteria of cfu and about 1 × 1010cfu/100kcal of the composition. In some embodiments, the amount of probiotic may be about 1 x 106cfu-about 1X 109cfu/100kcal of the composition. In addition, the nutritional composition may comprise a destabilized probiotic, and the final composition comprises some stabilized probiotic and some destabilized probiotic.
The nutritional composition may further comprise at least one prebiotic. The term "prebiotic" as used herein refers to a non-digestible food ingredient that exerts a health benefit to the host. Such health benefits may include, but are not limited to, selective stimulation of the growth and/or activity of one or a limited amount of beneficial gut bacteria, stimulation of the growth and/or activity of ingested probiotic (stabilized or destabilized) microorganisms, selective reduction of gut pathogens, and beneficial effects on gut short chain fatty acid profile. Such prebiotics may be naturally occurring, synthetic or developed by genetic manipulation of organisms and/or plants, whether such new sources are now known or later developed. Prebiotics may include oligosaccharides, polysaccharides, and other prebiotics that contain fructose, xylose, soy, galactose, glucose, and mannose. More specifically, prebiotics useful in the present disclosure may include lactulose, lactosucrose, raffinose, gluco-oligosaccharides, inulin, polydextrose powder, galacto-oligosaccharides, fructo-oligosaccharides, isomalto-oligosaccharides, soy oligosaccharides, lactosucrose, xylo-oligosaccharides, chito-oligosaccharides, manno-oligosaccharides, arabino-oligosaccharides, sialyl-oligosaccharides, fuco-oligosaccharides, and gentio-oligosaccharides, and combinations thereof.
In some embodiments, the total amount of prebiotic present in the nutritional composition may be from about 1.0g/L to about 10.0g/L of the composition (in liquid form). In certain embodiments, the total amount of prebiotics present in the nutritional composition may be from about 2.0g/L to about 8.0g/L of the composition.
The nutritional composition may comprise Polydextrose (PDX). If polydextrose is used as a prebiotic, in one embodiment, the amount of polydextrose in the nutritional composition may be in the range of about 1.0g/L to about 4.0 g/L. If polydextrose is used as a prebiotic, in one embodiment of the composition comprising stabilized probiotics, the polydextrose may be present in the nutritional product in an amount ranging from about 0.1mg/100kcal to about 0.5mg/100 kcal. In another composition, the amount of polydextrose may be about 0.3mg/100 kcal. In a preferred embodiment, at least 20% of the prebiotics should comprise Polydextrose (PDX).
In certain embodiments, the nutritional composition comprises galacto-oligosaccharides. The amount of galacto-oligosaccharides in the nutritional composition may be from about 0.2mg/100kcal to about 1.0mg/100 kcal. In other embodiments, the amount of galacto-oligosaccharides in the nutritional composition may be from about 0.1mg/100kcal to about 0.5mg/100 kcal. Galacto-oligosaccharides and polydextrose may also be supplemented to the nutritional composition in a total amount of about 0.6mg/100 kcal.
In some embodiments, the nutritional composition comprises an additional carbohydrate source, i.e., a carbohydrate source provided in addition to other carbohydrates described throughout the disclosure of the present invention. Suitable additional carbohydrate sources may be any used in the art, such as lactose, glucose, fructose, corn syrup solids, maltodextrin, sucrose, starch, rice syrup solids, and the like. The amount of additional carbohydrate in the nutritional composition may generally vary from about 5g to about 25g/100 kcal. In some embodiments, the amount of carbohydrate is from about 6g to about 22g/100 kcal. In other embodiments, the amount of carbohydrate is from about 12g to about 14g/100 kcal.
The nutritional composition may comprise a source of long chain polyunsaturated fatty acids (LCPUFAs) including docosahexaenoic acid (DHA). Other suitable LCPUFAs include, but are not limited to, alpha-linoleic acid, gamma-linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), and arachidonic acid (ARA).
In some embodiments, the nutritional composition may be supplemented with both DHA and ARA, and the weight ratio of ARA to DHA may be from about 1:3 to about 9: 1. In certain embodiments, the ARA to DHA ratio is from about 1:2 to about 4: 1.
The amount of long chain polyunsaturated fatty acids in the nutritional composition may vary from about 5mg/100kcal to about 100mg/100kcal, more preferably from about 10mg/100kcal to about 50mg/100 kcal.
Furthermore, the nutritional composition may be supplemented with an oil comprising DHA and ARA using standard techniques known in the art. For example, oils comprising DHA and ARA may be added to the nutritional composition by displacing the balance of an equivalent amount of the total fat blend typically present in the nutritional composition.
The source of DHA and ARA, if employed, may be any source known in the art, such as marine oils, fish oils, single cell oils, egg yolk lipids, and brain lipids. In some compositions, the DHA and ARA are derived from a single-cell Martek oil,Or a variant thereof. DHA and ARA may be in natural form, provided that the remainder of the LCPUFA source does not produce any significant deleterious effects on the infant. Alternatively, DHA and ARA may be used in purified form.
In one embodiment of the nutritional composition, the source of DHA and ARA is single cell oil, as taught in U.S. patent nos. 5,374,567, 5,550,156, and 5,397,591, the disclosures of which are incorporated herein by reference in their entireties.
In certain embodiments, the nutritional composition may be a milk-based nutritional composition that provides physiochemical and physiological benefits. As known in the art, milk proteins contain two main components: acid-soluble whey protein and acid-insoluble casein, the latter representing about 80% of the total protein content of milk. Upon entering the acidic environment of the stomach, casein precipitates and complexes with minerals, forming a semi-solid curd of varying size and firmness. Softer, smaller curd is more easily digested by the body than larger, harder curd. Curd formation can be an important consideration in the development of nutritional compositions including, but not limited to, infant formulas, medical foods, and preterm formulas. Thus, the stabilized probiotic may be combined with a composition comprising softer and smaller curd than standard infant formula.
One or more vitamins and/or minerals may also be added to the nutritional composition in an amount sufficient to supply the subject's daily nutritional needs. It will be appreciated by those of ordinary skill in the art that vitamin and mineral requirements will vary, for example, based on the age of the child. For example, an infant may have vitamin and mineral requirements that are different from those of children aged 1-13 years. Thus, this embodiment is not intended to limit the nutritional composition to a particular age group, but rather to provide a range of acceptable vitamin and mineral components.
The nutritional composition may optionally include, but is not limited to, one or more of the following vitamins or derivatives thereof: vitamin B1 (thiamine, thiamine pyrophosphate, TPP, thiamine triphosphate, TTP, thiamine hydrochloride, thiamine mononitrate), vitamin B2 (lactoflavin, flavin mononucleotide, FMN, flavin adenine dinucleotide, FAD, lactoflavin, riboflavin), vitamin B3 (niacin, nicotinic acid, nicotinamide adenine dinucleotide, NAD, nicotinic acid mononucleotide, NicMN, pyridine-3-carboxylic acid), vitamin B3-precursor tryptophan, vitamin B6 (pyridoxine, pyridoxal, pyridoxamine, pyridoxine hydrochloride), pantothenic acid (pantothenate, panthenol), folate (folic acid, folic acid (folacin), pteroylglutamic acid), vitamin B12 (cobalamin, methylcobalamin, desoxyadenosylcobalamin, cyanocobalamin, hydroxycobalamin, adenosylcobalamin), biotin, vitamin C (ascorbic acid), Vitamin A (retinol, retinyl acetate, retinyl palmitate, retinyl esters with other long-chain fatty acids, retinal, retinoic acid, retinol esters), vitamin D (calciferol, cholecalciferol, vitamin D3, 1,25, -dihydroxyvitamin D), vitamin E (alpha-tocopherol, alpha-tocopherol acetate, alpha-tocopherol succinate, alpha-tocopherol nicotinate, alpha-tocopherol), vitamin K (vitamin K1, phylloquinone, naphthoquinone, vitamin K2, menaquinone-7, vitamin K3, menaquinone-4, menaquinone-8H, menaquinone-9H, menaquinone-10, menaquinone-11, menaquinone-12, menaquinone-13), Choline, inositol, beta-carotene, and any combination thereof.
Furthermore, the nutritional composition may optionally include, but is not limited to, one or more of the following minerals or derivatives thereof: boron, calcium acetate, calcium gluconate, calcium chloride, calcium lactate, calcium phosphate, calcium sulfate, chloride, chromium chloride, chromium picolinate, copper sulfate, copper gluconate, fluoride, iron, carbonyl iron, ferric iron, ferrous fumarate, ferric orthophosphate, iron developers, polysaccharide iron, iodide, iodine, magnesium carbonate, magnesium hydroxide, magnesium oxide, magnesium stearate, magnesium sulfate, manganese, molybdenum, phosphorus, potassium phosphate, potassium iodide, potassium chloride, potassium acetate, selenium, sulfur, sodium, docusate sodium, sodium chloride, sodium selenate, sodium molybdate, zinc oxide, zinc sulfate, and mixtures thereof. Non-limiting exemplary derivatives of mineral compounds include salts, basic salts, esters, and chelates of any mineral compound.
Minerals may be added to the nutritional composition in the form of the following salts: such as calcium phosphate, calcium glycerophosphate, sodium citrate, potassium chloride, potassium phosphate, magnesium phosphate, ferrous sulfate, zinc sulfate, copper sulfate, manganese sulfate, and sodium selenite. Other vitamins and minerals known in the art may be added.
The nutritional compositions of the present disclosure may optionally include one or more of the following flavorings, including but not limited to flavoring extracts, volatile oils, cocoa or chocolate flavorings, peanut butter flavorings, cookie crumbs, vanilla or any commercially available flavoring. Examples of useful flavoring agents include, but are not limited to, pure anise extract, imitation banana extract, imitation cherry extract, chocolate extract, pure lemon extract, pure orange extract, pure peppermint extract, honey, imitation pineapple extract, imitation rum extract, imitation strawberry extract, or vanilla extract; or volatile oils, such as bee balm oil, bay oil, bergamot oil, cedarwood oil, cherry oil, cinnamon oil, clove oil, or peppermint oil; peanut butter, chocolate sauce, vanilla cookie crumb, butterscotch, toffee, and mixtures thereof. The amount of flavoring agent can vary widely depending on the flavoring agent used. The type and amount of flavoring agent known in the art can be selected.
The nutritional compositions of the present disclosure may optionally include one or more emulsifiers that may be added for stability of the final product. Examples of suitable emulsifiers include, but are not limited to, lecithin (e.g., from egg or soy), alpha-lactalbumin and/or mono-and diglycerides, and mixtures thereof. Other emulsifiers will be apparent to the skilled artisan and the selection of a suitable emulsifier will depend in part on the formulation and the final product. In some embodiments, the nutritional compositions of the present disclosure may comprise an emulsifier, such as citric acid esters of mono-and/or diglycerides, diacetyl tartaric acid esters of mono-and/or diglycerides, and/or octenyl succinic anhydride modified starch.
The nutritional compositions of the present disclosure may optionally include one or more preservatives, which may also be added to extend the shelf life of the product. Suitable preservatives include, but are not limited to, potassium sorbate, sodium sorbate, potassium benzoate, sodium benzoate, calcium disodium EDTA, and mixtures thereof.
The nutritional compositions of the present disclosure may optionally include one or more stabilizers. Suitable stabilizers for use in practicing the nutritional compositions of the present disclosure include, but are not limited to, acacia, ghatti, karaya, tragacanth, agar, furcellaran, guar gum, gellan gum, locust bean gum, pectin, low methoxyl pectin, gelatin, microcrystalline cellulose, CMC (sodium carboxymethylcellulose), methylcellulose hydroxypropyl methylcellulose, hydroxypropyl cellulose, DATEM (diacetyl tartaric acid esters of mono-and diglycerides), dextran, carrageenan, and mixtures thereof.
The nutritional compositions of the present disclosure may also include at least one additional plant nutrient, i.e., another plant nutrient component in addition to the pectin, starch, or other plant nutrient components described herein. The phytonutrients identified in human milk or derivatives, conjugated forms or precursors thereof are preferably comprised in the nutritional composition. For example, in some embodiments, the nutritional compositions of the present disclosure may comprise about 80 to about 300mg anthocyanins, about 100 to about 600mg proanthocyanidins, about 50 to about 500mg flavan-3-ols, or any combination or mixture thereof in 8 ounce (236.6mL) servings. In other embodiments, the nutritional composition comprises apple extract, grape seed extract, or a combination or mixture thereof. Furthermore, the at least one phytonutrient of the nutritional composition may be derived from any one or a blend of fruits, grape seeds and/or apple or tea extracts.
Examples of additional phytonutrients suitable for use in the nutritional composition include, but are not limited to, anthocyanins, proanthocyanidins, flavan-3-ols (i.e., catechins, epicatechins, etc.), flavanones, flavonoids, isoflavonoids, stilbenoids (i.e., resveratrol, etc.), proanthocyanidins, anthocyanidins, resveratrol, quercetin, curcumin and/or any mixtures thereof, and any possible combination of purified or natural forms of phytonutrients. Certain components, particularly plant-based components of the nutritional composition, may provide a source of plant nutrients.
The phytonutrient component of the nutritional composition may further comprise naringenin, hesperetin, anthocyanins, quercetin, kaempferol, epicatechin, epigallocatechin, epicatechin-gallate, epigallocatechin-gallate, or any combination thereof. In certain embodiments, the nutritional composition comprises from about 50 to about 2000nmol/L of epicatechin, from about 40 to about 2000nmol/L of epicatechin gallate, from about 100 to about 4000nmol/L of epigallocatechin gallate, from about 50 to about 2000nmol/L of naringenin, from about 5 to about 500nmol/L of kaempferol, from about 40 to about 4000nmol/L of hesperetin, from about 25 to about 2000nmol/L of anthocyanins, from about 25 to about 500nmol/L of quercetin, or mixtures thereof. Furthermore, the nutritional composition may comprise a metabolite of a phytonutrient or its parent compound, or it may comprise other kinds of edible phytonutrients, such as glucosinolates (glucosinolates) or sulforaphane. In certain embodiments, the nutritional composition comprises carotenoids, such as lutein, zeaxanthin, astaxanthin, lycopene, beta-carotene, alpha-carotene, gamma-carotene, and/or beta-cryptoxanthin.
The nutritional composition may further comprise isoflavonoids and/or isoflavones. Examples include, but are not limited to, isoflavone (genistin), daidzein (daidzin), glycitein, biochanin A, formononetin, coumestrol (coumestrol), irilone (irilone), tetrahydroxyisoflavone (orobol), pseudoindigoid (pseudoobtin), isocoumarin A and B (anagyroid isoflavanone A and B), calycosin (calycosin), glycitein, irigenin (irigenin), 5-O-methylisoflavone, pratensein (pratensein), prunetin (prunetin), psi-tectorigenin, hospholin (retusin), tectorigenin, irigenin (irigenin), formononetin (onone), puerarin, irilone (tectorone), deguelin (derrubinone), isoluteolin (isovaleryl), isovaleryl (irigenin), formononetin (formononetin), homocarinin (mangiferin-4-methylluteone), and homocarinin (mangiferin-4). Plant sources rich in isoflavonoids include, but are not limited to, soybean, psoralea, kudzu, lupin, fava bean, chickpea (chickpea), alfalfa, legumes and peanuts.
In one embodiment, the nutritional compositions of the present disclosure comprise an effective amount of choline. The effective amount of choline is from about 20mg choline per 8 ounce (236.6mL) serving to about 100mg per 8 ounce (236.6mL) serving. For example, 1 gram serving of the probiotic composition of the present invention is included per 8 ounce (236.6mL) serving of the nutritional composition as formulated. For example, 1L serving of the nutritional composition may be formulated to include 2-8 g serving of the probiotic composition of the present invention. For example, 1L serving of the nutritional composition may be formulated to include 3-6 g serving of the probiotic composition of the present invention.
The disclosed nutritional compositions may additionally comprise a source of beta-glucan. Glucans are polysaccharides, in particular polymers of glucose, which occur naturally and can be found in the cell walls of bacteria, yeasts, fungi and plants. Beta-glucans are themselves a diverse subset of glucose polymers, consisting of chains of glucose monomers linked together by beta-type glucosidic linkages to form complex carbohydrates.
Beta-1, 3-glucan is a carbohydrate polymer purified, for example, from yeast, mushrooms, bacteria, seaweed, or cereals. The chemical structure of beta-1, 3-glucan depends on the source of the beta-1, 3-glucan. Furthermore, various physicochemical parameters, such as solubility, main structure, molecular weight and branching, have an effect on the biological activity of β -1, 3-glucan.
Beta-1, 3-glucan is a naturally occurring polysaccharide with or without beta-1, 6-glucose or 1, 4-glucose branched side chains, found in the cell walls of a wide variety of plants, yeasts, fungi, and bacteria.
Beta-glucans derived from the baker's yeast Saccharomyces cerevisiae (Saccharomyces cerevisiae) consist of chains of D-glucose molecules linked by glucosidic bonds in the 1 and 3 positions with glucose side chains linked by glucosidic bonds in the 1 and 6 positions. Yeast-derived β -glucans are insoluble, fibrous, complex sugars having a linear general structure of glucose units having a β -1,3 main chain interspersed with β -1,6 side chains of typically 6-8 glucose units in length. More specifically, the beta-glucan derived from baker's yeast is poly (1,6) -beta-D-glucopyranosyl- (1,3) -beta-D-glucopyranose.
Moreover, beta-glucan is well tolerated and does not produce or cause excessive gas, bloating, or diarrhea in pediatric subjects. The addition of beta-glucan to a pediatric subject's nutritional composition (e.g., an infant formula, a growing-up milk, or another children's nutritional product) will improve the subject's immune response by increasing resistance to invading pathogens and thus maintaining or improving overall health.
The nutritional compositions of the present disclosure may comprise beta-glucan. In some embodiments, the beta-glucan is beta-1, 3; 1, 6-glucan. In some embodiments, β -1, 3; the 1, 6-glucan is derived from baker's yeast. The nutritional composition may comprise whole glucan particulate beta-glucan, PGG-glucan (poly 1, 6-beta-D-glucopyranosyl-1, 3-beta-D-glucopyranose), or any mixture thereof. In some embodiments, the particulate beta-glucan comprises beta-glucan particles having a diameter of less than 2 μm.
In some embodiments, the amount of beta-glucan present in the composition is from about 0.010 to about 0.080g/100g of the nutritional composition. In other embodiments, the nutritional composition comprises from about 10 to about 30mg of beta-glucan per gram serving. In another embodiment, the nutritional composition comprises from about 5 to about 30mg of beta-glucan per 8 ounce (236.6mL) serving. In other embodiments, the nutritional composition comprises beta-glucan in an amount sufficient to provide from about 15mg to about 90mg of beta-glucan per day. In some embodiments, the nutritional composition may be delivered in multiple doses to achieve a target amount of beta-glucan delivered to the subject throughout the day.
In some embodiments, the amount of beta-glucan in the nutritional composition is from about 3mg to about 17mg/100 kcal. In another embodiment, the amount of beta-glucan is about 6mg to about 17mg/100 kcal.
The nutritional composition may be directly excreted into the intestine of the subject. In some embodiments, the nutritional composition is excreted directly into the intestine. In some embodiments, the compositions may be formulated for enteral consumption or administration at the direction of a physician, and may be intended for targeted dietary management of diseases or conditions, such as celiac disease and/or food allergies, thereby establishing different nutritional needs through medical evaluation based on recognized scientific principles.
The nutritional compositions of the present disclosure are not limited to compositions comprising the nutrients specifically listed herein. Any nutrients may be delivered as part of the composition for the purpose of meeting nutritional needs and/or for optimizing the nutritional state in a subject.
The nutritional compositions of the present disclosure may be standardized to specific caloric content, they may be provided as a ready-to-use product, or they may be provided in a concentrated form.
In some embodiments, the nutritional composition of the present disclosure is a growing-up milk. Growing-up milk is a fortified milk-based beverage intended for children over the age of 1 year (usually 1-3 years, 4-6 years or 1-6 years of age). Growing-up milks are designed to be used as a supplement to diverse diets, providing additional support for children to obtain a sustained daily intake of all essential vitamins and minerals, macronutrients plus other functional dietary components (e.g., non-essential nutrients with alleged health promoting properties).
The exact composition of the nutritional compositions according to the present disclosure may vary between markets depending on local regulations and dietary intake information for the target population. In some embodiments, the nutritional compositions according to the present disclosure comprise a milk protein source, such as whole or skim milk, plus added sugar and sweeteners to achieve desired organoleptic properties, and added vitamins and minerals. Fat compositions are generally derived from milk raw materials. The indicator of total protein can be designed to match human milk, cow milk, or a lower limit. An indication of total carbohydrate is typically determined to provide as little sugar (e.g. sucrose or fructose) as possible to achieve an acceptable taste. Typically, vitamin a, calcium and vitamin D are added at levels that match the nutritional base values of native cow milk. Further, in some embodiments, a level of about 20% of the Dietary Reference Intake (DRI) or about 20% of the Daily Value (DV) per serving may be provided, with vitamins and minerals added. Moreover, nutrient values may vary from market to market, depending on the nutritional needs of the intended population, the base value of the raw materials, and regional regulations that have been determined.
In certain embodiments, the nutritional composition is hypoallergenic. In yet another embodiment, the nutritional composition is a non-genetically modified product. In one embodiment, the nutritional formulation is free of sucrose. The nutritional composition may also be free of lactose. In other embodiments, the nutritional composition does not comprise any medium chain triglyceride oil. In some embodiments, carrageenan is not present in the composition. In other embodiments, the nutritional composition does not contain all gums.
Thus, by practicing the present disclosure, stabilized probiotics are prepared with heretofore unrecognized stability. The stabilized bacterial mixture exhibits extremely high stability through the use of hydrolyzed mammalian proteins, particularly hydrolyzed mammalian proteins having over 70% peptides with a molecular weight of less than 2000 daltons. The stabilized probiotics are uniquely effective for nutritional applications at moderate humidity levels (e.g., water activity up to 0.4), where improved shelf life and stability in hot and humid environments is desirable. The stabilized probiotic may be packaged alone, or in combination with any of the embodiments of the nutritional compositions described herein.
The compositions prepared in each of examples 1-10 below were stored at 30 ℃ for 1 year, and the number of probiotics per 1g of the composition at 0 h and 1 year was determined for each composition, and the percentage of bacteria at 1 year of a given composition divided by the number of bacteria at 0 month of the composition multiplied by 100% was taken as the percentage of the composition remaining after 1 year of treatment at 30 ℃. The results show that the residual percentage of the total composition obtained in examples 1-7 is in the range of 79-88%, showing a higher survival rate of probiotics, for example the residual percentage of the total composition obtained in examples 1-5 is in the range of 81-85%; unfortunately, the residual percentages for all compositions obtained in examples 8-10 were in the range of 43-59%, for example, the residual percentages for all compositions obtained in example 10 were in the range of 45-51%, much lower than those obtained in a similar manner. In addition, the residual percentage of the probiotics in the nutritional composition obtained in example 12 was also determined in the manner of storage at 30 ℃ for 1 year as described above, and the results showed that the residual percentage of the nutritional composition was substantially the same as the results of the probiotic composition used therein (each with a difference of less than 2 percentage points), indicating that other substances such as protein source and the like do not affect the survival of the probiotics therein when mixed with the probiotic composition of the present invention.
Detailed Description
The present invention will be further described by the following examples, however, the scope of the present invention is not limited to the following examples. It will be understood by those skilled in the art that various changes and modifications may be made to the invention without departing from the spirit and scope of the invention. The present invention has been described generally and/or specifically with respect to materials used in testing and testing methods. Although many materials and methods of operation are known in the art for the purpose of carrying out the invention, the invention is nevertheless described herein in as detail as possible. The following examples further illustrate the invention without limiting it. In the following experiments, the probiotic compositions of the present invention were prepared in batches of at least 1000 grams at the time of actual dosing, per 1 gram weight of probiotic composition prescribed.
Example 1:probiotic composition (dry matter ingredient per 1 gram probiotic composition)
Probiotic (LGG): 1X 1010cfu,
Phospholipid (soybean lecithin): 10mg of the total weight of the mixture,
glycerol ester (lauric monoglyceride): 10mg of the total weight of the mixture,
oryzanol: 1mg of the total amount of the components,
zinc glycinate: 0.15mg of the total weight of the composition,
carbohydrate (lactose): add appropriate amount to 1 g.
The preparation method comprises the following steps: mixing probiotic (commercially available, semi-solid state) with oryzanol and zinc glycinate, adding phospholipid and glyceride, mixing, adding carbohydrate to the full amount of the prescription, stirring, mixing, and drying with hot air fluidized bed at 40 deg.C to water content below 4% to obtain granular or powdered probiotic composition.
Example 2:probiotic composition (dry matter ingredient per 1 gram probiotic composition)
Probiotic (BB 536): 1X 1011cfu,
Phospholipids (egg yolk lecithin): 5mg of the total weight of the mixture,
glycerol ester (stearic acid glycerol monoester): 20mg of the total weight of the mixture,
oryzanol: 0.5mg of the total weight of the composition,
zinc glycinate: 0.5mg of the total weight of the composition,
carbohydrate (sucrose): add appropriate amount to 1 g.
The preparation method comprises the following steps: mixing probiotic (commercially available, semi-solid state) with oryzanol and zinc glycinate, adding phospholipid and glyceride, mixing, adding carbohydrate to the full amount of the prescription, stirring, mixing, and drying with hot air fluidized bed at 40 deg.C to water content below 4% to obtain granular or powdered probiotic composition.
Example 3:probiotic composition (dry matter ingredient per 1 gram probiotic composition)
Probiotic (AH 1206): 1X 109cfu,
Phospholipid (soybean lecithin): 20mg of the total weight of the mixture,
glycerides (medium-chain acid diglycerides): 5mg of the total weight of the mixture,
oryzanol: 2mg of the total weight of the mixture,
zinc glycinate: 0.1mg of the total amount of the active ingredient,
carbohydrate (glucose): add appropriate amount to 1 g.
The preparation method comprises the following steps: mixing probiotic (commercially available, semi-solid state) with oryzanol and zinc glycinate, adding phospholipid and glyceride, mixing, adding carbohydrate to the full amount of the prescription, stirring, mixing, and drying with hot air fluidized bed at 40 deg.C to water content below 4% to obtain granular or powdered probiotic composition.
Example 4:probiotic composition (dry matter ingredient per 1 gram probiotic composition)
Probiotic (BB-12): 1X 1010cfu,
Phospholipids (egg yolk lecithin): 7.5mg of the total weight of the composition,
glycerides (medium chain acid triglycerides): 12.5mg of the total weight of the mixture,
oryzanol: 0.75mg of the total weight of the composition,
zinc glycinate: 0.25mg of the total amount of the active ingredient,
carbohydrate (maltodextrin): add appropriate amount to 1 g.
The preparation method comprises the following steps: mixing probiotic (commercially available, semi-solid state) with oryzanol and zinc glycinate, adding phospholipid and glyceride, mixing, adding carbohydrate to the full amount of the prescription, stirring, mixing, and drying with hot air fluidized bed at 40 deg.C to water content below 4% to obtain granular or powdered probiotic composition.
Example 5:probiotic composition (dry matter ingredient per 1 gram probiotic composition)
Probiotic (35624): 1X 1010cfu,
Phospholipid (soybean lecithin): 12.5mg of the total weight of the mixture,
glycerol ester (lauric monoglyceride): 7.5mg of the total weight of the composition,
oryzanol: 1.25mg of the total weight of the composition,
zinc glycinate: 0.1mg of the total amount of the active ingredient,
carbohydrate (fructose): add appropriate amount to 1 g.
The preparation method comprises the following steps: mixing probiotic (commercially available, semi-solid state) with oryzanol and zinc glycinate, adding phospholipid and glyceride, mixing, adding carbohydrate to the full amount of the prescription, stirring, mixing, and drying with hot air fluidized bed at 40 deg.C to water content below 4% to obtain granular or powdered probiotic composition.
In the above examples 1-5, the concentration of the carbohydrate in the dry matter of the probiotic composition is within the range of 17-76% due to the different bacterial concentrations in the probiotic raw material and the different feeding amounts.
Example 6:probiotic compositions
Ingredient ratios and methods were made with reference to examples 1-5, respectively, except that no carbohydrate was added to give 5 granular or powdered probiotic compositions, respectively.
Example 7:probiotic compositions
Compounding ratio and preparation reference is made to examples 1-5, respectively, except that the following binders (formulated as a 3% solution with water and added as a spray after the addition of the other materials) are also added separately for each formulation per 1 gram dry matter ingredient of the probiotic composition: 10mg of pectin, 7.5mg of sodium alginate, 20mg of chitosan, 12.5mg of carboxymethyl cellulose and 5mg of alginic acid to respectively obtain 5 granular or powdery probiotic compositions.
Example 8:probiotic compositions
The compounding ratio and the preparation method were respectively referred to examples 1 to 5 except that oryzanol was not added to obtain 5 compositions.
Example 9:probiotic compositions
The compounding ratios and preparation were made according to examples 1-5, respectively, except that zinc glycinate was not added to give 5 compositions.
Example 10:probiotic compositions
Compounding ratios and preparation methods were respectively referred to examples 1 to 5 except that oryzanol and zinc glycinate were not added to obtain 5 compositions.
Example 11:nutritional composition
The probiotic compositions obtained in examples 1-10 were separately packaged into empty capsules, sealed, and the resulting nutritional compositions were separately in the form of capsules. These nutritional compositions in capsule form can be added to a suitable food or beverage, for example to fruit juice or milk, at the time of consumption by the user.
Example 12:nutritional composition
The probiotic compositions obtained in examples 1 to 10 were mixed with commercially available infant formulas (A)Containing a protein source and also other lipids and other nutritional ingredients such as amino acids, vitamins, minerals etc.) to obtain a nutritional composition in the form of a formula, respectively, the probiotic composition comprising 0.1% (probiotic composition of example 1), 0.5% (probiotic composition of example 1), 1% (probiotic composition of example 1), 5% (probiotic group of example 1) of the nutritional compositionCompound) or 10% (probiotic composition of example 1) and 2% (probiotic composition of examples 6-10). For example, the amount of probiotic bacteria in these nutritional compositions in the form of a formula may be determined according to the actual situation, e.g. age group, race, etc. of the user, e.g. the nutritional composition in the form of a formula is prepared to contain 1 x 10 probiotic bacteria per 1 gram2-1×1010cfu。
All references cited in this specification, including without limitation all papers, publications, patents, patent applications, presentations, texts, reports, manuscripts, brochures, books, internet articles, journal articles, periodicals, and the like, are hereby incorporated by reference into this specification in their entirety. The discussion of the references herein is intended merely to summarize the assertions made by their authors and no admission is made that any reference constitutes prior art. Applicants reserve the right to challenge the accuracy and pertinency of the cited references.
Although embodiments of the present disclosure have been described using specific terms, devices, and methods, such description is for illustrative purposes only. The words used are words of description rather than limitation. It is to be understood that variations and modifications may be effected by one of ordinary skill in the art without departing from the spirit and scope of the disclosure as set forth in the appended claims. Additionally, it should be understood that aspects of the various embodiments may be interchanged both in whole or in part. For example, while methods for producing commercially sterile liquid nutritional supplements prepared according to those methods have been exemplified, other uses are also contemplated. Therefore, the spirit and scope of the appended claims should not be limited to the description of the preferred versions contained therein.
Claims (24)
1. A probiotic composition, comprising per 1 g: 1X 106-1×1014cfu probiotic bacteria, 5-20 mg phospholipids, 5-20 mg glycerides, 0.5-2 mg oryzanol, 0.1-0.5 mg zinc glycinate, 5-20 mg binder and a balance of carbohydrate, the probiotic composition being in the form of a granulate or powder and having a water content of less than 4%.
2. Probiotic composition according to claim 1, comprising 1 x 10 per 1g9-1×1011cfu of probiotic bacteria.
3. A probiotic composition according to claim 1, comprising 7.5-12.5 mg of phospholipids per 1 g.
4. A probiotic composition according to claim 1, comprising 7.5-12.5 mg of glycerides per 1 g.
5. A probiotic composition according to claim 1, wherein the carbohydrate is selected from the group consisting of polysaccharides, disaccharides and monosaccharides.
6. A probiotic composition according to claim 1, comprising between 0.75 and 1.25mg of oryzanol per 1 g.
7. A probiotic composition according to claim 1, comprising 0.1-0.25 mg of zinc glycinate per 1 g.
8. A probiotic composition according to claim 1, wherein said probiotic comprises viable microbial cells.
9. A probiotic composition according to claim 8, wherein said viable microbial cells comprise Lactobacillus rhamnosus.
10. A probiotic composition according to claim 8, wherein said viable microbial cells comprise Bifidobacterium longum BB536, Bifidobacterium longum subsp.infantis 35624, Bifidobacterium animalis subsp.lactis BB-12, or any combination thereof.
11. A probiotic composition according to claim 1, further comprising pectin.
12. A probiotic composition according to claim 1, said phospholipids comprising lecithin.
13. A probiotic composition according to claim 1, said glycerides comprising monoglycerides, diglycerides, triglycerides, or any combination thereof.
14. A probiotic composition according to claim 1, which is encapsulated in a capsule.
15. A probiotic composition according to claim 14, said capsule further comprising an amount of docosahexaenoic acid.
16. A probiotic composition according to claim 1, further comprising non-viable probiotic bacteria.
17. A probiotic composition according to claim 16, said non-viable probiotic being selected from the group consisting of: lactobacillus rhamnosus, bifidobacterium longum BB536, bifidobacterium longum subsp. infantis 35624, bifidobacterium animalis subsp. lactis BB-12, or any combination thereof.
18. A process for preparing a probiotic composition according to any one of claims 1 to 17, comprising the steps of:
(1) uniformly mixing the probiotics, the oryzanol and the zinc glycinate, and then adding the phospholipid and the glyceride for uniform mixing;
(2) adding powdery carbohydrate into the mixture, mixing, adding binder prepared with water, mixing, and making into wet particulate;
(3) drying the wet granules to reduce the water content to below 4%.
19. A nutritional composition, comprising:
the probiotic composition of any of claims 1-17; and
a protein source for providing nutrition to a user.
20. Nutritional composition according to claim 19, wherein the probiotic composition is present in an amount of 0.1-20% by weight of the total nutritional composition.
21. The nutritional composition according to claim 19, which is a pre-formulation of the probiotic composition with the protein source or which is a extemporaneous formulation of the probiotic composition with the protein source.
22. Nutritional composition according to claim 19, which is a powdered infant formula.
23. A nutritional composition according to claim 19, further comprising at least one additional lipid component.
24. The nutritional composition according to claim 19, further comprising a non-viable probiotic.
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CN108467846B (en) * | 2018-07-04 | 2019-02-15 | 中科宜康(北京)生物科技有限公司 | Probiotics solid beverage |
US20220000941A1 (en) * | 2018-11-28 | 2022-01-06 | Targedys | Mineral elements comprising hafnia alvei compositions |
KR102273663B1 (en) * | 2020-10-29 | 2021-07-05 | 주식회사 에치와이 | Coated probiotics, and food composition comprising the same and method for preparing thereof |
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