Folic acid and/or vitamin B12-lactoferrin complex

Food or edible material: processes – compositions – and products – Product with added vitamin or derivative thereof for...

Reexamination Certificate

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C426S074000, C530S400000, C424S439000, C514S904000, C514S905000

Reexamination Certificate

active

06500472

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a complex in which lactoferrin holds folic acid and/or vitamin B
12
(hereinafter referred to as folic acid and/or vitamin B12-lactoferrin complex), a method for the production thereof, and use thereof. A folic acid and/or vitamin B
12
-lactoferrin complex of the present invention is characterized in that folic acid in the complex exhibits improved photostability, acid tolerance and solubility as compared to folic acid present by itself, and vitamin B
12
in the complex exhibits improved acid tolerance as compared to vitamin B
12
present by itself, and accordingly is useful as a material for foods, drinks and medicines.
2. Description of the Related Art
Folic acid which was discovered as an antianemic factor a long time ago is one of the vitamins essential for the body. Further, various enzymes are known to require folic acid as a coenzyme. Also, folic acid is extensively involved in the metabolisms of nucleotides, glycine, histidine, or the like, and the biosyntheses of proteins. Pteroylglutamate, 7,8-dihydropteroylglutamate, or the like and polyglutamin compounds thereof are also known as folic acid in a broad sense, and all of these compounds have physiological activities as folic acid. A deficiency in folic acid is known to cause abnormal myelopoietic functions (megaloblastic anemia), nervous disorder, intestinal dysfunction, or the like.
Recently, the higher probability of newborns having abnormal neural tubes due to a nutrient deficiency in mothers during pregnancy having particularly attracted attention in Europe and the United States, it has been revealed that this risk can be reduced by administering folic acid (Czeizel, A. E., J. Pediat. Gastroenterol. Nutr., vol. 20, pp. 4-16, 1995). Furthermore, it has been reported that an increase in homocysteine, which is associated with heart diseases, and a depletion of folic acid in the serum occur simultaneously (Jacob, R. A., M. M. Wu, S. M. Henning & M. E. Swendseid, J. Nutr., vol. 124, p. 1072, 1994). It has been also reported that folic acid exerts a preventive effect on cancer, particularly epithelial cancer (Glynn, S. A., D. Albanes, Nutr. Cancer, vol. 22, p. 101, 1994). Thus, the importance of folic acid is being recognized again.
On the other hand, folic acid is known to be relatively heat stable in the absence of oxygen. However, folic acid has low heat stability and storage stability in the presence of oxygen. For example, about 60% of folic acid was reported to be lost in cow's milk by heat sterilization or during storage (Renner E., Japanese Journal of Dairy & Food Science, vol. 35, p. A121-A135, 1996), and 25% of folic acid was lost in powdered milk (Oamen E. E., Hansen, A. P. M. & Swartzel, K. R., J. Dairy Sci., vol. 72, pp. 614-619, 1989). Furthermore, it is also known that folic acid is extremely sensitive to and rapidly decomposes in light and must be handled in the dark (Henderson B. G., Annu. Rev. Nutr., vol. 10, pp. 319-335, 1990).
The presence of a folic acid-binding protein in cow's milk is conventionally known (Ford, J. E., D. N. Salter & K. J. Scott, J. Dairy Res., vol. 36, p. 435, 1969). This folic acid-binding protein has a molecular weight of about 25 kDa, binds one molecule of folic acid, and is suggested to promote absorption from the intestinal tract (Said H. M., F. K. Ghishan & R. Redha, Am. J. Physiol., vol. 252, p. G229, 1987). However, the content of this folic acid-binding protein in milk is as low as 10 mg/L (Parodi P. W., Diet & Health News for New Zealand Health Professionals, vol. 27, pp. 1-4, 1998). Further, there has been no report on the photostability of this folic acid-binding protein.
As described above, folic acid has lately attracted considerable attention because of its important physiological functions, and the use of folic acid as a material for foods, drinks and medicines is desirable. However, there exists various limitations; for example, it is difficult to use folic acid, particularly in drinks, because of its poor photostability and solubility, which occasionally causes precipitation or requires shielding in packaging. Furthermore, as for the folic acid-binding protein, its content in milk is small and the outlook for its industrial scale production is still in doubt.
On the other hand, since vitamin B
12
is associated with the synthesis of heme, which is necessary for erythropoiesis, its deficiency is known to generate megaloblasts and cause anemia. Furthermore, since vitamin B
12
is essential for the generation and growth of cells, its deficiency causes inflammation of mucosal tissues, diarrhea, or the like. Vitamin B
12
is also known to have important roles in the reproductive functions and nervous system (Toru Fukui, “How to Take Vitamins and Minerals”, Maruzen, 1997). Vitamin B
12
deficiency is attributed mainly to a vegetarian diet or often to gastrocectomy or abnormal absorption from the intestinal tract. Two extrasecretal proteins are known to be associated with vitamin B
12
absorption, i.e., haptocorrin secreted from the salivary gland (Toyoshima S., H. Saido, F. Watanabe, K. Miyatake and Y. Nakano, Abstract of XV International Congress of Nutrition, 204, 1993) and the intrinsic factor secreted from the stomach (Levine J. S., P. K. Nakane and R. H. Allen, Gastroenterology 79, 493, 1980); but the absorption mechanism is complicated. The recommended daily intake of vitamin B
12
for an adult in Japan is set to a low 2.4 &mgr;g. If this vitamin B
12
intake is obtained from food products, loss by cooking has to be taken into consideration. Furthermore, the recommended intake of vitamin B
12
per day for an adult in the United States, i.e., Optimal Daily Allowance (ODA), is set as 10 to 300 &mgr;g. A more positive intake of this vitamin will be required.
The presence of proteins which bind to vitamin B
12
in cow's milk has been suggested (Peter, W. P., Australian J. Dairy Tech., 53, 37-47, 1998). However, the whole picture of vitamin B
12
binding proteins has not yet been revealed. Vitamin B
12
-binding bovine serum albumin (BSA) is the only one known protein (U.S. Pat. No. 4,082,738) and is being used in the quantification of erythrocytes (Japanese Patent Publication S57-5005281).
Further, vitamin B
12
is known to be relatively heat stable but not acid tolerant (Owen R. Fennema, Food Chemistry 3nd ed., Dekker, N.Y.). Accordingly, there is a need for a method to improve the stability, in particular the acid tolerance, of vitamin B
12
which has useful physiological functions.
Thus, both folic acid and vitamin B
12
are effective substances to ameliorate anemia or the like, but problems in stability restricts their use to limited areas.
SUMMARY OF THE INVENTION
In the present invention, it was newly found that lactoferrin interacts with folic acid and vitamin B
12
to form a folic acid and/or vitamin B
12
-lactoferrin complex. Moreover, the folic acid in the complex has increased photostability and drastically increased solubility and is even stable in acid, as compared to folic acid present by itself, and vitamin B
12
in the complex has increased acid tolerance as compared to vitamin B
12
present by itself. Thus, the present invention has been completed. Accordingly, an object of the present invention is to provide a folic acid and/or vitamin B
12
-lactoferrin complex in which folic acid and/or vitamin B
12
are incorporated into lactoferrin and a method for producing the same. Another object of the present invention is to provide medicines or foods and drinks prepared by using the folic acid and/or vitamin B
12
-lactoferrin complex.
Lactoferrins to be used in the present invention can be those derived from milk, blood or the like. The source materials are not particularly restricted and can be derived from humans, bovines, hogs, or the like. Lactoferrins obtained by gene recombination can also be used. These lactoferrins can be those purified or partially purified, or materials having a low purity, such as WPC and skimmed milk powder. These lactoferrins

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