Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for...
Reexamination Certificate
2002-01-30
2004-05-04
Lilling, Herbert J. (Department: 1651)
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
C435S071100, C435S194000, C435S212000, C435S221000, C435S232000, C435S252310, C426S061000
Reexamination Certificate
active
06730504
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a food product containing a thrombolytic enzyme, nattokinase, but containing little or none of a blood coagulation factor, vitamin K2.
2. Description of the Prior Art
The
Bacillus natto
was discovered by Sumi et al. to produce the thrombolytic enzyme nattokinase (Experientia Vol. 43, p. 1110 (1987)), and thus the nutritional value of natto and its value as a health food are looked at again. It is known that nattokinase itself acts as a fibrinolytic enzyme, and lyses thrombi when ingested as a food. Nattokinase has extremely good characteristics such as having a long half life and retaining its effectiveness for a long period of time. In addition, it is reported that urokinase having the same thrombolytic activity as nattokinase does not clearly have a therapeutic effect on incipient central retinal vein occlusion, but that this effect can be clearly seen with nattokinase (Nishimura et al., Ganka Rinsho Ihou (in Japanese), Vol. 88, No. 9, pp. 53-57 (1994)).
Accordingly, food products containing large amounts of nattokinase, for example, powders and capsules formed from
Bacillus natto
culture extract, are being marketed as health foods.
On the other hand, the
Bacillus natto
is known to produce large amounts of vitamin K2. Vitamin K2 is known as an essential component of the blood coagulation system. Further, vitamin K2 has other physiological functions, and is said to cause absorption disorders in newborns and osteoporosis in the elderly when it is deficient, and said to cause hemolytic anemia, splenomegaly, nephropathy, hepatopathy and the like when it is present in excess. Thus, a
Bacillus natto
culture extract simultaneously contains nattokinase as a thrombolytic system effector and vitamin K2 as a blood coagulation system effector.
The necessary daily intake of vitamin K2 for adults is generally 55 to 65 &mgr;g. Large amounts of vitamin K's are found in foods such as seaweed, broccoli and the like. Because vitamin K's are also produced by enterobacteria and by
Bacillus natto
that grow in the intestines when natto is ingested, it is possible for vitamin K2 to be produced. Therfore, it is said that the necessary intake of vitamin K2 can be satisfied by a normal diet. Thus, it is generally not necessary to supplement vitamin K2.
On the other hand, problems occur with patients who are administering synthetic inhibitors of the vitamin K-dependent blood coagulation factors (for example, protothrombin VII, IX, X and the like) to prevent thrombosis. When these patients ingest a natto or a
Bacillus natto
culture extract containing the thrombolytic enzyme, nattokinase, in order to prevent thrombosis and the like, they also ingest vitamin K2 at the same time, and the effect of synthetic inhibitor of the vitamin K-dependent blood coagulation factor is counteracted.
Accordingly, in order to prevent thrombogenesis, a food product of
Bacillus natto
culture extract in which the vitamin K2 content has been reduced are desired, and therefore, methods of reducing vitamin K2 in the food product have been attempted. Using organic solvents such as hexane to extract fat-soluble vitamin K2 is one of these methods.
However, the problems with this method are: fat-soluble nutrients are also extracted and removed in addition to vitamin K2: the need to remove organic solvents such as hexane leads to problems in terms of a production technique such as an increase in the cost of production; consumer opposition to the use of organic solvents; and the possibility that organic solvents remain in the food.
Accordingly, a
Bacillus natto
culture extract in which the vitamin K2 content has been reduced is in demand, as well as a method in which vitamin K2 can be simply and easily extracted without using organic solvents.
SUMMARY OF THE INVENTION
The present invention was put into practice for the purpose of solving the aforementioned problems, and provides a
Bacillus natto
culture extract containing nattokinase and an amount of vitamin K2 below a specified level, and a method for producing the same. According to the present invention, the problems with the food products containing nattokinase of the prior art are solved. Therefore, an optimal diet that nutrients are excellent, the activity of nattokinase is enhanced, and there are no longer any concerns about the excessive intake of vitamin K2, is provided. Furthermore, because little or none of the blood coagulation factor, vitamin K2, is present, an optimal diet for patients with diseases of the blood coagulation system is also provided.
The present invention relates to a
Bacillus natto
culture extract containing nattokinase and 1 &mgr;g or less of vitamin K2/g dry weight (hereinafter referred to as the
Bacillus natto
culture extract of the present invention).
In a preferred embodiment, the
Bacillus natto
culture extract of the present invention is obtained by a method which comprises culturing natto in a liquid culture medium, and treating the culture medium obtained therefrom with chitosan.
In a preferred embodiment, the
Bacillus natto
culture extract is in the form of a concentrated extract, a paste, a powder, a granule, a capsule, a drinkable preparation (beverage) or a tablet.
In addition, the present invention relates to a method for producing a
Bacillus natto
culture extract containing nattokinase and 1 &mgr;g or less of vitamin K2/g dry weight, comprising culturing natto in a liquid culture medium to obtain a cultured medium, and treating the cultured medium with chitosan.
Further, the present invention relates to a method for collecting vitamin K2, comprising treating a sample containing vitamin K2 with chitosan, and extracting the vitamin K2 adsorbed by the chitosan with an organic solvent.
According to the present invention, a
Bacillus natto
culture extract containing nattokinase and a concentration of 1 &mgr;g or less of vitamin K2/g dry weight can be obtained by a simple and easy method using chitosan. Because it contains an exceedingly small amount of vitamin K2, this
Bacillus natto
culture extract can prevent thrombogenesis and the excessive intake of vitamin K2 in healthy individuals, and is an excellent food product that allows people who limit their intake of vitamin K2 to safely ingest nattokinase.
These and other advantages of the present invention will become apparent to those skilled in the art upon reading and understanding the following detailed description.
DETAILED DESCRIPTION OF THE INVENTION
The
Bacillus natto
culture extract of the present invention contains nattokinase and 1 &mgr;g or less of vitamin K2/g dry weight. Preferably, it contains 0.5 &mgr;g or less of vitamin K2/g dry weight, and more preferably contains 0.1 &mgr;g or less of vitamin K2/g dry weight.
The form of the
Bacillus natto
culture extract includes a culture medium itself obtained by culturing the
Bacillus natto
, completely removing the bacteria from the liquid culture medium, and almost completely or completely removing the vitamin K2 from the culture medium. It is possible for the
Bacillus natto
culture extract to be in the form of a liquid, a powder, or a solid. For example, a liquid can include a concentrated extract of the culture medium and a paste. A powder can include a dried powder, a granule obtained by further drying a concentrated extract or a paste, and the like. For example, a solid can be a tablet obtained by compressing. The tablet can be coated depending on a variety of uses, including a sugar-coated tablet. Alternatively, the concentrated extract, paste, powder or granule can be contained in a capsule. Further, it can be in the form of a drinkable preparation, such as a beverage.
The
Bacillus natto
culture extract of the present invention contains nattokinase. The nattokinase content is not limited, however, it is preferable to be 20 FU/g or more, more preferably 1000 FU/g or more, and even more preferably 2500 FU/g or more. These are variable according to concentration of the
Bacillus natto
culture extrac
Amin & Turocy LLP
Japan Bio Science Laboratory Co., Ltd.
Lilling Herbert J.
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