Pigment- containing materials to be added to feeds

Details Pigment- containing materials to be added to feeds Pigment- containing materials to be added to feeds

2001-05-23

2004-03-16

Saucier, Sandra (Department: 1651)

Drug, bio-affecting and body treating compositions

Preparations characterized by special physical form

Food or edible as carrier for pharmaceutical

C435S067000, C435S252100

Reexamination Certificate

active

06706278

ABSTRACT:

TECHNICAL FIELD
The present invention relates to a pigment-containing substance for feed additives consisting of a microorganism culture precipitate which contains carotenoid compounds.
BACKGROUND ART
“Carotenoid compounds” is a generic name for a group of pigment which have a long-chain polyene structure, the majority of which have a C40 tetraterpenoid, and present yellow, orange, red or purple color. More specifically it refers to the compounds such as &bgr;-carotene, astaxanthin, canthaxanthin, zeaxanthin, echinenone, adonirubin and adonixanthin. These compounds can be used as natural pigments useful for feed additives, food additives, pharmaceuticals etc. For example, astaxanthin is valuable from an industrial point of view as a feed additive such as a color improver for bred fishes such as salmon, trout, red sea bream etc., and also as a safe natural food additive. Adonixanthin is as promising as astaxanthin, as a feed additive, a food additive and pharmaceutical as the astaxanthin is, if its industrial production process can be established. Canthaxanthin has been used as a feed additive, food additive, cosmetic, etc., and zeaxanthin has been used as a feed additive and food additive. Further, echinenone and adonirubin also expected to be used as feed additives, food additives etc. As production processes for these carotenoid compounds, chemical synthesis, production by microorganisms, extraction from natural products etc. are known, and regarding astaxanthin and canthaxanthin, their chemical synthetic products are already on sale. Comparing with a chemical synthesis, the production process of carotenoid compounds by microorganisms is advantageous due to the high level of safety, since neither metal catalysts nor solvents are used therein. Generally, carotenoid compounds are unstable in the presence of oxygen and light. However, since carotenoid compounds produced by microorganisms are accumulated inside the cell of a microorganism, their stability is ensured by the cell membrane, cell wall and other antioxidants. When carotenoid compounds are used as feed additives or the like, a high level of storage stability is an advantage, and if the microorganism itself can be used, this would be a great advantage. In contrast, if intercellular carotenoid compounds are used after being extracted and purified, it is necessary to used chlorine-containing solvents or the like which may be unsafe. Accordingly, the use of a microorganism culture precipitate consisting of a microorganism which contain carotenoid compounds as a feed additive has a great advantage. In the case where the microorganism producing carotenoid compounds has a hard cell wall, the absorption efficiency is extremely low if a natural cell is provided. So the cell should be disrupted mechanically or decomposed with chemical agents or enzymes, and this leads to a cost disadvantage. In addition, there is a problem that the stability of carotenoid compounds decreases after the cell is disrupted. Furthermore, when the content of carotenoid compounds existing in a microorganism culture precipitate is low, a large mount of precipitate is needed in the production process of the feed, and this causes other problems such as poor operability, high transport cost and nutritional inbalance of the feed. However, there is not known a pigment-containing substance consisting of a microorganism whose lack of a hard cell wall allows easy use of the accumulated carotenoid compounds; and, further consisting of a microorganism culture precipitate which contains at least 3 mass % carotenoid compounds.
Among carotenoid compounds, astaxanthin is a red pigment contained in Pisces such as salmon, trout, red sea bream etc., and Crustacea such as shrimps, crabs, etc., and it is useful because of its beautiful color, and as stated above, it has been broadly used as a feed additive and a natural food additive. Regarding the red yeast
Phaffia rhodozyma
, which is known to produce astaxanthin, there is a problem that, being a yeast, it has a hard cell wall. As processes for the production of astaxanthin by bacteria, the followings are known, but regarding each of them, the content per weight of dry cell is low: a bacterium
Brevibacterium
No.103 strain belonging to the genus
Brevibacterium
produces merely 0.003% of astaxanthin per weight of dry cell (
Journal of General and Applied Microbiology
, 15, 127, 1969). Also, another bacterium
Paracoccus marcusii
DSM11574 strain belonging to the genus
Paracoccus
produces only 0.022% of astaxanthin per weight of dry cell (WO 99/6586).
The object of the present invention is to provide a pigment-containing substance for feed additives consisting of a microorganism culture precipitate which contains a high concentration of carotenoid compounds useful as a natural pigment.
DISCLOSURE OF THE INVENTION
In order to achieve the above object, the present invention provides the following means.
1. A pigment-containing substance for feed additives comprising a microorganism culture precipitate which contains at least 3 mass % carotenoid compounds.
2. The pigment-containing substance for feed additives, characterized in that at least 40 mass % of the carotenoid compounds is astaxanthin.
3. The pigment-containing substance for feed additives, characterized in that a DNA nucleotide sequence corresponding to 16S ribosomal RNA of the microorganism in the microorganism culture precipitate has at least 98% homology with the nucleotide sequence shown in SEQ ID NO: 1.
4. The pigment-containing substance for feed additives, characterized in that the microorganism in the microorganism culture precipitate is E-396 strain or a mutant, thereof.
This specification includes part or all of the contents disclosed in the specification of Japanese Patent Application No. 11-279337, which is a priority document of the present application.
The present invention is described more specifically as follows.
The pigment-containing substance for feed additives of the present invention comprises a microorganism culture precipitate. First, the microorganism culture precipitate is disclosed. The microorganism culture precipitate of the present invention can be obtained by a process wherein mainly the cell of microorganism producing carotenoid compounds is cultured to produce these compounds, the resulting culture is treated by filtration, centrifugation etc., followed by the removal of a certain amount of moisture. Any method for culturing a strain which produces carotenoid compounds can be used on the condition that carotenoid compounds are generated. For example, the following method can be adapted. As a medium, one which contains the followings is used: carbon sources, nitrogen sources, inorganic salt, which are necessary for the growth of bacteria producing carotenoid compounds, and particularly required substances (e.g. vitamins, amino acids, nucleic acid bases etc.) as needed. The carbon sources to be used include carbohydrate such as glucose, sucrose, fructose, trehalose, mannose, mannitol and maltose; organic acids such as acetic acid, fumaric acid, citric acid, propionic acid, malic acid and malonic acid; alcohols such as ethanol, propanol, butanol, pentanol, hexanol and isobutanol; and the like. The ratio of added carbon sources depends on the type of carbon sources, but generally 1 to 100 g per L of medium, preferably 2 to 50 g is applied. The nitrogen sources to be used include one or more selected from potassium nitrate, ammonium nitrate, ammonium chloride, ammonium sulfate, diammonium hydrogen phosphate, ammonia, urea and the like. The ratio of added nitrogen sources depends on the type of nitrogen sources, but generally 0.1 to 10 g per L of medium, preferably 1 to 3 g is applied. The inorganic salt to be used includes one or more selected from potassium dibydrogen phosphate, dipotassium hydrogen phosphate, disodium phosphate, magnesium sulfate, magnesium chloride, ferrous sulfate, ferrous chloride, manganous sulfate, manganous chloride, zinc sulfate, zinc chloride, cupric sulfate, calcium chloride, calcium carbonate, sodium c

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