TW201033359A - Alga strain rich in DHA and application of the same - Google Patents

Abstract

The present invention relates to an alga strain rich in DHA and application of the same. The alga of the present invention is easy in production, rich in DHA and DHA is easily purified. The products manufactured from this alga containing DHA with high quality which can be used as human healthy foods and pharmaceutics, additives of aquatic and livestock product, and fish meal.

Description

201033359 * VI. Description of the invention: 'Technical field to which the invention pertains>> The present invention relates to a strain of algae containing high-quality DHA, and further to the present invention relates to a strain of algae which is simple in fatty acid composition and easy to purify DHa. [Prior Art] docosahexaenoic acid (DHA), a polyunsaturated fatty acid containing six double bonds, is an essential fatty acid and is a major component of the cell membrane in brain and god L tissues. DHA is known to have functions that promote brain and retina development, as well as prevent central nervous system, menstrual disease, and cardiovascular disease. At present, the health foods on the market in Xuxi are advertised as containing DHA, which emphasizes the prevention or improvement of depression, Alzheimer's disease and cardiovascular diseases. In addition, it can also be used as a health supplement for pregnant women and young children. Department development. For all advanced countries today, almost all of them are faced with the problem of aging and declining. The excellent DHA products can meet these problems and create unlimited business opportunities. At present, there are many products of polyunsaturated fatty acids (including DHA) on the market. The fish oil is often taken from large fishes, so it is also known as deep sea fish oil. Since humans and most animals cannot produce DHA by metabolic methods, they must be ingested from the outside. However, the use of fish oil to provide DHA may have the following disadvantages, including unstable DHA content, complex fish oil composition, and high cost of refining DHA. May contain heavy metals and polychlorinated biphenyls and other toxic substances, which may cause fetal malformation, or affect the development of infants and young children; in addition, fish oil is a foraging property that is not accepted by vegetarians and has a distinct astringency. In addition, a sufficient amount of DHA should be added to aquatic feed and livestock feed to promote the development and growth of the culture while enhancing its resistance to disease. From 3 201033359, the current scale of aquaculture and livestock farming has gradually increased, and the DHA of fish source that it relied on in the past has already been in short supply. The algae source DHA provides a reliable source. At present, there are also some algae species used to produce DHA, but some difficulties have also been encountered. For example, DHA has low productivity, high mass production cost, and the fatty acid composition of algae species is complicated, and it is not easy to further extract high-purity DHA. & Grypf/iecodim'um eo/mii &

The algal 楂±^ΌΉΑ of Schizochytrium sp., wherein the DHA content of the Schizochytrium Jt product is about 18-22% of the dry algae weight, which has the advantage of being cheap, and the disadvantage is that the DHA content of the product is not easy to obtain; It is about 15-20%, which has the advantage of easily refining high-purity DHA. However, the DHA content is low and it is not easy to culture. Therefore, the mass production cost is high, and at the same time, because of the lack of alcohol, the algae powder of m'ww is recognized. It has a strong pungent smell, so it must be processed by some food processing before it can be sold, causing its cost to soar. ^ Therefore, the development of a high-capacity, easy-to-purify source of DHA will help develop DHA products and provide higher quality D DHA products at a lower price for people to purchase. SUMMARY OF THE INVENTION In view of the lack of prior art, the object of the present invention is to develop a high-yield, easy-to-purify DHA source to reduce the cost and price of DHA products, and to facilitate the promotion of commodities. In view of the above, the present invention provides a medicinal strain for producing DHA, particularly a strain of algae, which has been deposited in the Food Industry Development Research Institute, which is BCRC980009. The invention also includes a use of the algae strain 201033359 for the production of DHA, in particular, the _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Industrial Development Research Institute 'The algae plant registration number is BCRC98〇〇〇9. The present invention also finds that the algae strain is rich in dha. It can be used for processing human health foods and health medicines, aquatic or animal product additives, and juvenile fish feed. Therefore, the present invention also includes an algal plant product, which is A DHA semi-finished product or purified product obtained by drying or extracting the algae plant described in claim 1 of the patent application. The algae oil obtained by using the algae strain of the invention has the following advantages over the fish oil: high DHA content, stable quality, simple ingredients, easy to further refine into high purity OHA, no heavy metals and polychlorinated biphenyls, etc. Toxic ingredients, vegetarians are edible, and there is no fishy smell. In addition, compared with other algae, the use of the algae strain of the present invention for mass production of similar products has the following advantages: no pungent taste, high DHA content, and the algae strain is native to Taiwan, produced in Taiwan. There are no problems with exotic species. φ [Embodiment] During the development of the present invention, microbial screening was carried out for the coastal areas of Taiwan. ^ There are 16 strains of Thraustochytrium (Family).

The single-celled organism of Thraustochytriaceae has the potential to be a source of DHA. These single-celled organisms are cultured under standard growth conditions, ie 400 mL of medium is contained in a soaring serum bottle containing three components, peptone, The yeast extract and glucose are 1 g, 2 g and 4 g per liter, respectively. The culture base has a salinity of 30 ppt. When inoculated, the inoculation amount is 5 g per liter. When the algae cells are collected two weeks after inoculation, the dry weight of each algae cell can reach 30~180 mg/L, and the lipid content per gram of dry algae cells is 15〇~6〇〇 post, 201033359 and DHA content accounts for 1〇%~58% of total fatty acid content. Among them, the DHA content of the algae strain, such as the 咕 咕 特别 特别 特别 特别 特别 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋 疋The algae strain has a lipid content of up to 73% under appropriate conditions, of which 40% is DHA. Therefore, the obtained algae cells can be directly dried to prepare a dry powder product containing 29% DHA or the oil can be extracted to prepare an oily product containing 4% DHA, or further refined by 〇1£4. Come out and make DHA products. Since the fatty acid composition of this strain is very simple (Fig. 2), except for DHa, it contains almost only palmitic acid (molecular structure of 16 carbon saturated acid, C16:0) which is very different from the physical properties of DHA, so it can be used simply. The process is separated from the palmitic acid to obtain high purity DHA. ',', the present inventors have found that the genus strain is rich in DHA, and therefore the present invention also encompasses the use of the genus Algae to produce a DHA-rich article. The aforementioned strain of the genus is a 乂μγ(10) valerian strain. Currently registered in the Food Industry Development Institute's registration number BCRC980009. < Oral The algae products currently on the market are usually obtained by a drying or extraction process. Therefore, the present invention also encompasses an algal plant product obtained by drying or extracting the above-mentioned algae strain. The products produced are based on manufacture and use, including but not limited to dry powder products, oily products or DHA-rich products. It will be readily understood that the processing of the algal strain of the present invention to produce a strain of algae can be carried out by various processing techniques known in the art, such as cold drying or various extraction steps and apparatus, as exemplified by U.S. 6,750,048, 5,492,938 Patent No. 'European Union 17〇7〇61 Patent^Technology, etc.' However, the present invention is characterized by the discovery of a novel strain of algae, the use of the strain of the algae, and the product of the strain of the algae, and the means for producing the product of the algae may take any Known 201033359 technology is not the key to the invention. The following embodiments are intended to further understand the advantages of the present invention and to limit the scope of the claims of the present invention. Example: Culture of the algae strain of the present invention. The toxicity analysis type of the model is carried out in seawater of 30 ppt salinity, and contains 1, 2, 4, and 8 g of digested protein, yeast extract, glucose, and agar. When the solid medium of L is cultured in above, the appearance of the algal strain BL10 of the present invention is white or yellowish (Fig. A). In the early stages of colony growth, slid-type amoeboid cells (Fig. B, arrows) were observed. The steamed cells are mixed with 1 liter of serum bottle and cultured with 100 mL of medium. The initial medium is formulated as digested protein, yeast extract and glucose, and the content is divided into 3 g, 6 g and 14 g per liter. Salinity is 1 〇ppt. The inoculated algae strain was previously deposited in the Food Industry Development Research Institute, and the algae strain numbered BCRC980009 was inoculated at a rate of 0.005 g per liter. Pumping and oscillating (150 rpm) during the culture. Glucose was digested 3.5 days after inoculation, and the amount of dip was 50 g per liter. Thus, the culture density on the 6th day after inoculation reached 59 〇g per liter, and the total fatty acid and Dha content reached 73 J; 〇 and 29 分别, respectively. The fatty acid analysis profile obtained by gas chromatography mass spectrometry with Supelco SP-2380 (81§1^-Shi also 11'1^,1^0,1;8) pipe column is shown in the second figure. , wherein 1_6 is a fourteen carbon saturated acid 201033359 (C14:0), a fifteen carbon saturated acid (ci5: 〇), a sixteen carbon saturated acid (C16:0), an eighteen carbon saturated acid (C18:0), Docosapentaenoic acid (C22: 5n-6), and DHA (C22: 6n-3). Comparison of DHA content As shown in Table 1, the biomass weight and DHA content of the BL10 strain cultivated by the method of sputum production are compared with the algae containing DHA in other literatures. Table 1 Comparison of DHA content of algae strains and other algae of the present invention (g/L > DHA(g/L) CR* Reference domain Aurantiochytritm BL10 59 16.8 8.4% Schizochytrium G13/2S 63 6.1 3.9% of the present invention Ganuza et αί, 2008 Schizochytrium N-2 13 2.7 3.9% Kamlangdee and Fan, 2003 Schizocf^irium S31 6 0.3 1.3% Wnetal.,2005 Schizochytrium SR21 35 4.2 3.9% Yokochi et αί, 1998 Thraustochytrium ATCC34304 5 0.5 1.9% Bajpd et al 1991, Thraustochytrium ONC-T18 26 4-5 6.4% Buija et al.7 2006 Crypthecodinium ATCC30772 109 19.0 2.2% Swaaf ei a/., 2002 * CR: conversion rate of medium-DHA (conversionrate)

It is known from the present table that the algae species of the present invention not only have a higher DHA capacity, but also have a better conversion rate (CR), that is, when the same amount of medium is consumed, the amount of DHA that can be obtained is higher, The cost required for mass production of the same weight DHA is lower. Acute toxicity test The method of the acute toxicity test is to dry the algal powder obtained by freeze-drying the algae cells obtained by the above culture, and mix it with water and then pour it into the stomach of the SD rat. The control group was filled with the same volume of water. Six male and female rats in the experimental group and the control group had an average body weight of 250 g and 180 g, respectively. Feeding dose 8 201033359 is 8 grams per kilogram per rat. Two weeks after the feeding, abnormalities such as weight loss and diarrhea occurred. , or whether there is death, the obvious results show that the feeding of dry algal flour does not cause any regular = dose 'that is, 8 grams per kilogram per rat for two rats; - A good dha mass production of high-capacity and easy-to-purify algae strains to obtain helmets; a type of fine ship that does not cause foreign problems; products: Shiben low, Kang, medicine and thin Industry needs. In addition, the replacement of fish oil with steam oil by steaming plants will help the human race to maintain the balance of marine ecology. All other features disclosed in this specification may be replaced by the same, equal or similar purpose characteristics, so Features other than those described in the specification are merely examples of the features. The present invention has been described above with reference to the preferred embodiments thereof, and is not intended to limit the invention, and various modifications and changes may be made without departing from the spirit and scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a view showing the appearance of bli〇 of the vine plant of the present invention in white, color or micro-steam. The first panel B shows the sliding amoeba-type cells (arrows) in the early stage of cell population growth of BL10. 201033359 The second figure is the fatty acid analysis of BL10, where 1-6 are 14 carbon saturated acid (C14_.0), 15 carbon saturated acid (C15:0), 16 carbon saturated acid (C16:0), 18 carbon saturated. Acid (C18:0), 22 carbon 5-enoic acid (C22:5n-6), and DHA (C22:6n-3). 201033359 References

Bajpai P. K.5 Bajpai Ρ. and Ward Ο. P.5 199. Optimization of production of docosahexaenoic acid (DHA) by Thraustochytrium aureum ATCC 34304. J. Am. Oil. Chem. Soc., 68: 509-514.

Buija AM, Radianingtyas H., Windust A. and Barrow CJ, 2006. Isolation and characterization of polyunsaturated fatty acid producing Thraustochytrium species: screening of strains and optimization of omega-3 production. App. Microbiol. Biotechnol., 72: 1161-1169 .

And the Az.

Kamlangdee N. and Fan K. W., 2003. Polyunsaturated fatty acid production by Schizochytrium sp. isolated from mangrove. Songklanakarin J. Sci. TechnoL, 25: 643-650.

SwaafM· E. D·, Sijtsma L. and Pronk J. T., 2002· High-cell-density fed-batch cultivation of the docosahexaenoic acid producing marine alga Crypthecodinium cohniL Biotech. Bioeng·, 81: 666-672.

Wu S, T·, Yu S, T· and Lin L·广 P., 2005 · Effect of culture conditions on docosahexaenoic acid production by Schizochytrium sp. S31. Process Biochem., 40: 3103-3108.

Yokochi T., Honda D., Higashihara T. and Nakahara T., 1998. Optimization of docosahexaenoic acid production by Schizochytrium limacinum SR21. AppL Microbiol. Biotechnol., 49: 72-76.

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Claims (1)

201033359 VII. Patent application scope: 1. A strain of algae for producing DHA-rich products. 2. 2. The strain of the plant described in claim 1 is Aurantiochytrium mangrovei f. 3. The strain of algae as described in item 2 of the patent application is deposited by the Food Industry Development Research Institute under the accession number BCRC980009 or its derived curtain. 4. A use of a Jwrflwhoctyi Www strain of algae for the production of a DHA-rich product. 5. The use according to claim 4, wherein the aforementioned Au Cong ti〇chytrium genus is Auranti 〇 M Mumma 叩 ei steamed. 6. The use according to item 5 of the patent application, wherein the aforementioned "Acacia" is a strain of algae or a derivative thereof of the number BCRC980009 registered by the Food Industry Development Research Institute. 7. An algae plant product which will be prepared by drying or extracting the patented patent. The back of the first paragraph of the dry circumference 8. If the application is in the scope of the patent range 帛i, the algae product, the oily product or the DHA-rich product. - The system is dry and unproductive 9. The semi-purified DHA product of the algae plant as described in claim 1 of the patent application. Ππ,,, is purified or 12