201233339 六、發明說明： 【發明所屬之技術領域】 血==關於一種可作為健康輔助食品等而簡單攝取之 錢性疾錢时品，更詳細而種包含特定之 Μ生物之醱酵物作為有效成分 【先前技術】 成刀的血检性疾病預防食品。 根據日本厚生勞動省發表之按傷病分類 包含高血塵性疾病、缺血性 晉縻費 戾病及細血官疾病之循環系統 之疾病的診療醫療費最高，占診療醫療費整體之212%: 並且’该等疾病均參與血管中之血栓之異常形成。 ★於血液中，有凝固系統與纖溶系統之2種之作用。若血 管壁損傷，則血小板凝聚’從而引起一次止血，其後作為 凝固系統因子之凝血酶於血液血纖維蛋白原中活動，藉此 :成灰纖維蛋白而完成止血。另一方面，自血管内皮二胞 刀泌之作為纖溶系統因子的組織纖維蛋白溶酶原活化因子 (tPA ’ t丨ssue piasmin〇gen⑽丨補叫將存在於血液中之作為 酶源的纖溶酶原轉換為纖維蛋白溶酶，該纖維蛋白溶酶使 士此於血官内形成之血纖維蛋白分解。已知凝固系統與纖 冷系統之平衡之崩潰成為腦梗塞或心肌梗塞等血栓性疾病 之原因，一般認為新的纖溶系統亢進物質之開發可較大有 助於該等疾病之預防、治療。 本發明者專人取得關於將藉由培養納豆菌類而產生之酶 或微里成分進行可逆性低分子化直至穩定之狀態之物質之 製造方法的專利權（專利文獻1)。 161203.doc 201233339 [先前技術文獻] [專利文獻] [專利文獻1]日本專利第3902015號公報 【發明内容】 [發明所欲解決之問題] 種藉由使tPA活性亢進並且引 而預防血栓性疾病的功能性 本發明之目的在於提供一 起自血管内皮細胞釋出tPA， 食品。 [解決問題之技術手段] 本發明者等人發現：於該課題之下，添加使納豆菌類產 生之酶或微量成分進行可逆性低分子化直至穩定狀態之低 分子肽成分於血管内皮培養細胞令，結果培養液中之斤八 活性亢進’又’使小鼠口服攝取’結果血液中之心活性 提高，從而完成本發明。 即，本發明係提供 Π] 一種血拴性疾耗防食品，其包含如τ成分作為有 效成分·將利用㈣酶水解㈣而成之糖化物作為培養基 用基材’於其中添加酵母萃取物而製備_用培養基 於遠培養基中接種作為納豆菌之枯草桿菌Ακ(寄存編號： FERM Ρ 18291)，使㈣酵及熟成之後，分 狀成分； 王城之凌 [2] 如上述丨& 4入 J之血栓性疾病預防食品，其令 pH值4.5〜6.5、28〜32。 晖係衣 z L之條件下進行2個月以上者； [3] 如上述[η或m J 4〖2]之灰栓性疾病預防食品，其中熟 I61203.doc 201233339 係於_4.0〜6·0、13〜17°C之條件下進行4個月以上者； ⑷如上述U]至附任-項之血栓性疾病預防食品， 其中血栓性疾病係選自由深部靜脈血栓症、門靜脈血检 症、腎靜脈血栓症、頸靜脈血栓症、布加氏症候群、腋_ 鎖骨下靜脈血栓症、腦靜脈竇血栓症及肺血检检塞症所組 成之群中之1種或超過旧的靜脈血检症，或選自由腦梗 塞、心肌«、腸間膜㈣血栓症、下肢急性動脈血检 症、肝動脈血栓症、腎動脈血检症、脾動脈血检症及閉塞 性動脈硬化症所組成之群中之i種或超…種的動脈血检 症；以及 [5]如巾任—項之血栓性疾病預防食品， 其相對於食品整體之重量，含有〇 〇5〜⑽重量％之有效成 分。 [發明之效果] 根據本發明，可提供—種於日常生活中可簡單地口服攝 取之血拴性疾病預防食品。 【實施方式】 於本發明之食品中作為有效成分而含有之成分係將利用 展粕酶水解包含玉米澱粉之澱粉而成之糖化物作為培養基 用基材，於其中添加作為氮源之酵母萃取物，而製備醱酵 用坨養基，於s亥培養基中，接種作為納豆菌之枯草桿菌 AK(寄存編號：FERM P-I8291)，使其酸酵及熟成之後， 分餾所生成之液狀成分。 此處，用作醱酵用培養基之基材之糖化物可使用利用澱 I6I203.doc 201233339 粉酶水解自玉米種子分離、並純化之玉米澱粉而成者，亦 可使用粗純化之玉米澱粉而代替經純化者。又， … 粉以外，可使用水解大豆粉或米糠或該等之混合物而成者 作為培養基用基材。 用作醱酵用培養基之氮源之酵母萃取物可使用將消化啤 酒酵母（Saccharomyces cerevisiae Meyen)之菌體並萃取 水/谷性成分乾燥而成者等通常於細菌培養時作為氮源而添 力σ者。 於本發明之食品之有效成分之製造所使用的醱酵用培養 基中’除上述糖化物及酵母萃取物以外，可視需要調=蛋 白質等有機物或無機鹽類等》作為有機物，可列舉大豆蛋 白質或其他植物性蛋白質，作為無機鹽類，可列舉氣化 約、氣化納、構酸鈉等。 又，較佳為亦對於作為培養基用基材之利用澱粉酶水解 澱粉而成之糖化物進而添加糖分，就該等糖分而言，例如 可列舉嚴糖（砂糖）、葡萄糖（gluc〇se)、始糖等。 接種於如上所述製備而成之醱酵用培養基之醱酵菌係作 為納豆菌之枯草桿菌（Bacillus subtilis)AK，且括草桿菌 AK係以「（寄存編號）FERM p_18291」寄存於曰本獨立行 政法人產業技術综合研究所。又，視所希望，除枯草桿菌 AK以外，亦可接種不抑制枯草桿菌Ακ之增殖之作為乳酸 才干菌之乳桿菌（Lactobacillus)或作為乳酸球菌之鏈球菌 (strept〇coccus)、酵母（Sacchar〇myces cerevisiae)或綠麴菌 (Aspergillus oryzae)之菌體 '菌萃取物或菌醱酵萃取物。 161203.doc 201233339 進而’於本發明之食品之有效成分之製造所使用的醱酵用 培養基中，根據要求，可添加不抑制枯草桿菌Ακ之增殖 之白菜、捲心菜、胡蘿蔔、藥用人蔘、洋芹、旱芹、洋蔥 萼之植物萃取物。 括草桿菌ΑΚ係以如下方式而獲得：對通常之作為納豆 菌之枯草桿菌附上紫外線、χ射線照身十、高低溫環境 d〇〇°c、〇。〇、與乳酸菌之競爭、容易製作芽胞之培養基 =·〇重量％之肉萃取物、1G.G重量％之蛋白腺、5 Q重量❶之 氣化鈉、15.0重量％之瓊脂、65.0重量％之蔬菜(捲心菜、 胡蘿«、旱；f、洋；f)壓榨汁]等各種條件，而對所發現之 耐藥菌進行反覆繼代培養並選拔。 以上述方式獲得之枯草桿菌Ακ菌株係且 菌學性質。 ㈣-有如下所述之 U)形態學之性質 1 細胞之形狀及大小 才干菌 1.0〜1 ·2 X3 ·0~50 μπι 2 有無細胞之多形性 無 3 有無運動性 有 （周毛性之鞭毛） 4有無芽胞 有橢圓菌體之大致中央 (b)培養性質 1肉汁環脂平板培養 161203.doc 201233339 圓形集群白色混濁 2 肉汁液體培養 上部或下部菌凝體 (C)生化學性質 1革蘭氏（gram)染色陽性 2 硝酸鹽之還原陽性 3 MR(methyl red，甲基紅）試驗陰性 4 VP(Voges-Proskauer，伯波二氏）試驗陽性 5吲哚之生成陰性201233339 VI. Description of the Invention: [Technical Fields of the Invention] Blood == A kind of money-based money that can be easily ingested as a health supplement, etc., and more specifically contains a specific sputum as a active ingredient. [Prior Art] A blood test disease prevention food for a knife. According to the Ministry of Health, Labor and Welfare issued by the Ministry of Health, Labor and Welfare, the medical treatment costs of diseases including high bloody diseases, ischemic sputum and sputum diseases are the highest, accounting for 212% of the total medical expenses: And 'these diseases are all involved in the abnormal formation of blood clots in the blood vessels. ★ In the blood, there are two kinds of functions of the coagulation system and the fibrinolytic system. If the blood vessel wall is damaged, platelet aggregation is caused to cause a hemostasis, and then thrombin, which is a factor of the coagulation system, acts in the blood fibrinogen, thereby completing hemostasis by forming gray fibrin. On the other hand, tissue plasminogen activator (tPA ' t丨ssue piasmin〇gen (10), which is a fibrinolytic system factor, is a fibrinolytic enzyme that is present in the blood as an enzyme source. The zymogen is converted to plasmin, which decomposes fibrin formed in the blood. It is known that the collapse of the balance between the coagulation system and the fibrillation system becomes a thrombotic disease such as cerebral infarction or myocardial infarction. For this reason, it is generally believed that the development of new fibrinolytic system-promoting substances can greatly contribute to the prevention and treatment of such diseases. The inventors have obtained a reversible effect on the enzyme or micro-ingrown components produced by culturing natto bacteria. Patent of the method for producing a substance which is in a state of being low-molecularized to a stable state (Patent Document 1). 161203.doc 201233339 [Prior Art Document] [Patent Document] [Patent Document 1] Japanese Patent No. 3902015 [Problems to be Solved by the Invention] The function of preventing thrombotic diseases by inducing tPA activity and promoting it is to provide a blood vessel together [Technical means for solving the problem] The inventors of the present invention have found that under this subject, a low molecular molecule which reversibly decomposes an enzyme or a trace component which is produced by natto bacteria to a stable state is added. The peptide component is cultured in a vascular endothelium, and as a result, the activity in the culture medium is increased, and the oral activity of the mouse is increased, and the cardiac activity in the blood is increased, thereby completing the present invention. That is, the present invention provides a blood sample. An environmentally-friendly food, which contains, for example, a τ component as an active ingredient, and a saccharide obtained by (4) enzymatic hydrolysis (IV) as a substrate for a medium, wherein a yeast extract is added thereto, and is prepared by inoculating the medium with a medium. Bacillus subtilis Α κ (registered number: FERM Ρ 18291), after (four) leaven and mature, the fractal component; Wang Chengzhi [2] such as the above 丨 & 4 into J thrombotic disease prevention food, which makes the pH Values of 4.5 to 6.5, 28 to 32. Those who have been exposed to the conditions of liters of clothing L for more than 2 months; [3] such as the above [η or m J 4 〖2] gray sputum disease prevention food, which is cooked I61203. Doc 201233339 is carried out under the conditions of _4.0~6·0, 13~17 °C for more than 4 months; (4) The above-mentioned U] to attached-term thrombotic disease prevention food, wherein the thrombotic disease is selected from the deep part Venous thrombosis, portal vein blood test, renal vein thrombosis, jugular vein thrombosis, Budd-Chiari syndrome, 腋_ subclavian vein thrombosis, cerebral venous sinus thrombosis, and pulmonary blood test 1 or more than the old venous blood test, or selected from cerebral infarction, myocardial «, intestinal mucosa (four) thrombosis, lower extremity acute arterial blood test, hepatic artery thrombosis, renal artery blood test, splenic artery blood test and occlusion An arterial blood test of i or super species in a group consisting of atherosclerosis; and [5] a thrombotic disease prevention food such as a towel, which contains 〇〇5~(10) relative to the weight of the whole food. % by weight of active ingredient. [Effects of the Invention] According to the present invention, it is possible to provide a bloody disease prevention food which can be easily taken orally in daily life. [Embodiment] The component contained in the food of the present invention as an active ingredient is a saccharide obtained by hydrolyzing a starch containing corn starch using a sputum enzyme as a substrate for a medium, and a yeast extract as a nitrogen source is added thereto. In the shai culture medium, the Bacillus subtilis AK (registered number: FERM P-I8291) was inoculated as a bacterium, and the liquid component formed by fractionation was carried out after acid fermentation and aging. Here, the saccharide used as the substrate for the fermentation medium can be obtained by using the corn starch separated and purified from the corn seed by the hydrolysis of the lake I6I203.doc 201233339, or by using the crude purified corn starch instead. Purified. Further, in addition to the powder, a hydrolyzed soybean powder or rice bran or a mixture of the above may be used as the substrate for the medium. The yeast extract used as a nitrogen source for the fermentation medium can be obtained by using a cell which digests the yeast of Saccharomyces cerevisiae Meyen and extracting water/gluten components, and is usually used as a nitrogen source during bacterial culture. σ. In the fermentation medium used for the production of the active ingredient of the food of the present invention, in addition to the above-mentioned saccharide and yeast extract, an organic substance such as a protein or an inorganic salt may be adjusted as needed, and examples thereof include soybean protein or Other plant proteins include, as an inorganic salt, gasification, gasification, sodium hydride, and the like. In addition, it is preferable to further add a sugar to the saccharide which is obtained by hydrolyzing starch with an amylase as a substrate for a medium, and examples of the sugar include, for example, sugar (granulated sugar) and glucose (glucose). Start sugar and so on. The yeast strain inoculated in the fermentation medium prepared as described above is used as Bacillus subtilis AK of Bacillus natto, and the Bacillus subtilis AK is deposited in Sakamoto by "(registered number) FERM p_18291" Administrative Corporation Industrial Technology Research Institute. Further, as long as it is desired, in addition to Bacillus subtilis AK, Lactobacillus as a lactic acid-producing bacterium or Strept〇coccus or a yeast (Sacchar〇) which does not inhibit the proliferation of Bacillus subtilis Ακ can be inoculated. Myces cerevisiae) or Aspergillus oryzae's bacterial extract or bacterial extract. 161203.doc 201233339 Further, in the fermentation medium used for the production of the active ingredient of the food of the present invention, cabbage, cabbage, carrot, medicinal aphid, and ocean which do not inhibit the proliferation of Bacillus subtilis Ακ can be added as required. A plant extract of parsley, celery, and onion. The bacterium of the genus Bacillus is obtained by attaching ultraviolet rays to the Bacillus subtilis which is usually natto bacteria, sputum rays, and high and low temperature environments d〇〇°c, 〇. 〇, competition with lactic acid bacteria, medium for easy production of spores = 〇% by weight of meat extract, 1G.G% by weight of protein gland, 5 Q by weight of sodium sulphate, 15.0% by weight of agar, 65.0% by weight Vegetables (cabbage, carrots, drought, f, ocean; f) pressed juice] and other conditions, and the drug-resistant bacteria found in the subculture and selection. The Bacillus subtilis Ακ strain obtained in the above manner has bacteriological properties. (4) - U) Morphological properties as described below 1 Cell shape and size Talents 1.0~1 · 2 X3 · 0~50 μπι 2 With or without cell morphism No 3 With or without motility (perimeter hair Flagella) 4 with or without spores in the approximate center of elliptic cells (b) culture properties 1 succulent ring fat plate culture 161203.doc 201233339 round cluster white turbidity 2 gravy liquid culture upper or lower bacterium (C) biochemical properties 1 leather Positive gram staining 2 Reductive positive for nitrate 3 MR (methyl red) test negative 4 VP (Voges-Proskauer, Bobo II) test positive 5 吲哚 negative
6硫化氫之生成陰性 7檸檬酸之利用陽性 8觸媒陽性 9 生長之範圍 pH 值 5.5 〜7.0 溫度25°C〜40°C 枯草桿菌AK菌株係根據要求’與包含乳酸菌之其他酿 酵菌一併於上述醱酵用培養基上，以約4·5〜6·52ρΗ值、 約28〜3 2 °C之條件下進行2個月以上之醱酵。推測於低於該 等區域之pH值區域或溫度區域中，即便進行2個月以上之 醱酵’本發明所使用之特定之納豆菌亦未高效率地使糖及 氮源同化，而未充分獲得所得之tPA釋出亢進物質所期待 之效果。另一方面’於高於上述區域之pH值區域或溫度區 域中，由於醱酵並不充分’從而特定之納豆菌未高效率地 使糖及氮源同化，而未充分獲得所得之tPA釋出亢進物質 I6l203.doc 201233339 所期待之效果。 繼醱酵後’菌類係於相同培養基上，以約4 〇〜6〇之ρΗ 值、約13〜17t之條件下進行4個月以上之熟成。推測於低 於該等區域之pH值區域或溫度區域中’即便進行4個月以 上之熟成，作為醱酵產物之具有各種活性之胺基酸、脂蛋 白、脂多糖（lipo P〇lysaccharide)、脂質等亦未充分低分子 量化，而未充分獲得所得之tPA釋出亢進物質所期待之效 果。另一方面，推測於高於上述區域之pH值區域或溫度區 域中，活性下降，而與上述同樣地未充分獲得所得之tpA 釋出亢進物質所期待之效果。 於分館經酿酵及熟成階段而生成之液狀成分時，可採用 過據或離^分離等眾所周知之分離方法，所錢之食品用 原液可直接，或進行濃縮或稀釋而用作本發明之血检性疾 病預防食品之有效成分。 =為本發月之血检性疾病預防食品之有效成分，例如可 調配由Enzamin研究所股份有限公司製造之Εηζ_η原液 (ENM)或作為其2G倍濃縮萃取物之Enzamin濃縮液（ENM_ HL) 〇 因此，作為以上述方式獲得之有效成分之構成成分，含 有用以使有機體内酶合六 成變今易之物質，即可逆性切割藉 由職酵而獲得之酿你Λ 人 從而作為活性胺基酸殘基之片段，此外 δ有如腺嘌呤、i;過。入 ^ m m ?、胞嘴Π定、胸腺p密β定、尿啦σ定之類 可於有機體内活用之有 ’用物質。一般認為此種有用物質係 下培養遽液：包含別朴s丨冰卜 J錄列德卡（Besredka)所提倡之抗病 '61203.doc 201233339 毒（Antivirus)的組織活性因子；費拉托夫（Filat〇v)所說明 之生命源刺激素’且藉由生物化學性反應將該等組合，以 安全且有效地起作用之方式進行處理。 包含此種有效成分之本發明之血栓性疾病預防食品藉由 刺激血液之纖溶作用之生命現象，而適當調節凝固系統與 纖溶系統之平衡，藉此預防血栓性疾病之發病。 可藉由本發明之血拴性疾病預防食品而預防之血栓性疾 病只要為起因於血栓而產生之疾病，則並無特別限定，例 如可列舉.選自由深部靜脈血栓症、門靜脈血栓症、腎靜 栓症頸靜脈血栓症、布加氏症候群、腋-鎖骨下靜 脈血栓症、腦靜脈竇血栓症及肺血栓栓塞症所組成之群中 之靜脈血栓症，或選自由腦梗塞、心肌梗塞、腸間膜動脈 血栓症 '下肢急性動脈血栓症、肝動脈血栓症、腎動脈血 栓症、脾動脈血栓症及閉塞性動脈硬化症所組成之群中之 動脈血栓症等。 本發明之血栓性疾病預防食品可將以上述方式製備而成 之有效成分與食品領域中慣用之賦形劑（例如，澱粉或糊 f、纖維素、乳糖、麥芽糖、還原乳糖、還原麥芽糖、山 梨糖醇、#露糖醇、赤藻糖醇、木糖醇等)或輔助劑(例 如’溶劑、分散介質、被覆劑、穩定劑、稀釋劑、保存 1防腐齊|、殺菌劑、抗真菌試劑、等滲透壓試劑、吸收 抑制試劑' 崩解劑、乳化劑、結合劑、潤滑劑、色素等) 進行混合，並藉由食品領域中慣用之製劑方法，而製成例 如片劑、膠囊、顆粒 '粉末、萃取液、溶液、浆液、懸浮 1612〇3.d〇( 201233339 液、乳濁液之形態。 本發明之血拴性疾病預防食品可含有以 ★ 於食品整體之·§·吾队 y、本身換算相對 體之重量，約0·05〜晴量 重量％、更佳為約】〜85重 佳為約°.1， %、芒社生从 進而較佳為約5〜8〇重吾 最佳為約I 〇〜5〇重量％兹 ϊ 有效成分。 “之生成之液狀成分作為 乍為本發明之其他態樣，亦關於一種用以製 血栓性疾病預防舍σ 從 &上逑 w “ 利用㈣酶水解包含玉米殺粉之 歲粉而成之糖化物作為培養基絲材，於其中添加作為^ 源之酵母萃取物”備_料養基，並於該培養基中接 種作為納旦菌之枯草桿菌AK(寄存編號：FERM P 助υ，使其酿酵及熟成之後，分傲所生成之液狀成分的 成分之用途；-種用以製造血栓性疾病預防食品之括草桿 菌ΑΚ之用途；特徵為攝取血栓性疾病預防食品之血检性 疾病之預防或治療方法。 以下，基於實施例對本發明進行更詳細之說明，但本發 明並不受該等實施例限定。 [實施例1] 於2_3 kg之黃玉米澱粉、〇.5 kg之大豆蛋白脒、〇 5 kg之 米糠汁、80 g之氯化鈣、150 g之食鹽中加入5〇 kg之純化 水’加熱並進行溶解。繼而使其冷卻，並加入5〇 g之殿粉 酶而使其充分糖化。糖化結束後，加入1.5 kg之砂糖、1 · 5 kg之葡萄糖（glucose)、450 g之酵母萃取物（曰本製藥 (股））、1.5 kg之米飴、80 g之填酸鈉、5 kg之蔬菜之壓榨 161203.doc 201233339 汁（捲心菜、胡蘿蔔、旱斧、，、篆甘 早斥羊芹）及純化水，而使總重量 為 150 kg 。 並且，添加氫氧化納而將PH值調整為7.2〜7.6之範圍 内’將其放人培養罐中，於12代下高壓滅菌20分鐘。使 ,、冷卻後帛種枯草桿菌Ακ株，並於溫度士，C之恆溫 室中，於pH值4.5〜6.5之條件下使其醱酵6()天，繼而於溫 度15 士 2 C之& /m至中，於阳值4』〜6 〇之條件下使其熟成 12〇天。於下對其上清液進行2Q分鐘滅菌，使其自然 放置冷卻而使培養㈣明化。以過濾紙將其㈣後，藉由 檸檬酸將pH值調整為3.5_37，進而於95<t下殺菌後，於 90°C以上之溫度下裝入5加侖之罐中。以上述之方式而獲 得125升之液狀食品用原液（enm)。 將100 g之所獲得之食品用原液（ENM)中之一般分析結果 示於以下之表1中。 又，關於上述食品用原液，使用T〇s〇h公司製造之管柱 (TSKgel G2500PWXL)，並利用使移動相為水、乙腈及三 氟乙酸之55 : 45 ·· 0.1混合液的液體高速層析圖（Sh〇dex公 司製造：GPC SYSTEM-21)而測定尺寸排除層析（SEC， size exclusion chromatography)，將此時之檢測器感度（紫 外分光光度計：mV)與分子量已知之標準品之溶出時間進 行比較’將分析之分子量分佈示於圖1，又，將於該圖中 之分子量餾分之面積占整體之比例（百分率）示於表1。 [表η 分子量範圍 峰面積百分率（％) 161203.doc 12 201233339 10,000以上 微量 3,000-10,000 2 1,000-3,000 9 500-1,000 9 500以下 80 合計 100 繼而，對所獲得之食品用原液（ENM)中含有之低分子物 質是否由於熱而產生改質或分解進行評價。 藉由使用TSK gel G2500PWXL管柱（Tosoh股份有限公司 製造）之尺寸排除層析法，對於121°C下加熱30分鐘之ENM 的分子量分佈進行測定。並且，以同樣之方式對未置於高 溫中之同批次的ENM(對照ENM)之分子量分佈進行測定。 將該等之結果示於表2。 [表2] 分子量範圍 10,000以上 3.000- 10,000 1.000- 3,000 500-1,000 500以下 合計6 Hydrogen sulfide generation negative 7 Citric acid utilization positive 8 Catalyst positive 9 Growth range pH 5.5 ~ 7.0 Temperature 25 ° C ~ 40 ° C Bacillus subtilis AK strain according to requirements 'with other bacteria containing lactic acid bacteria The fermentation was carried out for 2 months or more on the above-mentioned fermentation medium at a temperature of about 4·5 to 6·52 ρ and about 28 to 32 ° C. It is presumed that in the pH region or the temperature region below the region, the specific natto used in the present invention does not efficiently assimilate the sugar and nitrogen sources, even if it is carried out for two months or more. The effect obtained by the obtained tPA releasing the entangled substance is obtained. On the other hand, 'in the pH region or temperature region above the above region, the fermentation is not sufficient', so that the specific natto bacteria do not efficiently assimilate the sugar and nitrogen sources, and the obtained tPA is not fully obtained. The effect expected by the material I6l203.doc 201233339. After the fermentation, the fungus is cultured on the same medium, and is aged for 4 months or more at a pH of about 4 〇 to 6 、 and about 13 to 17 tons. It is presumed that in the pH region or temperature region below the region, even if it is matured for more than 4 months, the amino acid, lipoprotein, lipopolysaccharide (lipo P〇lysaccharide) having various activities as a fermentation product, Lipids and the like are also not sufficiently low in molecular weight, and the effect obtained by the obtained tPA releasing the entangled substance is not sufficiently obtained. On the other hand, it is presumed that the activity is lowered in the pH region or the temperature region higher than the above region, and the effect expected from the obtained tpA releasing drug is not sufficiently obtained as described above. When the liquid component formed by the fermentation and the ripening stage is used in the branch, a well-known separation method such as separation or separation can be used, and the used food liquid can be used as the present invention directly or concentrated or diluted. The active ingredient of food for prevention of blood test diseases. = The active ingredient of the blood-detecting disease prevention food for this month, for example, the Εηζ_η stock solution (ENM) manufactured by Enzamin Research Institute Co., Ltd. or the Enzamin concentrate (ENM_ HL) as its 2G-concentrated extract 〇 Therefore, as a constituent component of the active ingredient obtained in the above manner, a substance for analysing the enzyme in the organic body is changed, and the material obtained by the artificial leaven can be reversely cut to obtain the active amine group. A fragment of an acid residue, in addition to δ such as adenine, i; In the case of ^ m m ?, sputum sputum, thymus p-density β, urinary sputum, etc., can be used in the organism. It is generally considered that this kind of useful substance is a culture sputum: it contains the tissue activity factor of anti-pathology '61203.doc 201233339 toxic (Antivirus) advocated by Besredka; (Filat〇v) The described life-source stimulators' and combined by biochemical reactions are treated in a safe and effective manner. The thrombotic disease preventing food of the present invention containing such an active ingredient can prevent the onset of a thrombotic disease by appropriately adjusting the balance between the coagulation system and the fibrinolytic system by stimulating the life phenomenon of fibrinolysis of blood. The thrombotic disease which can be prevented by the bloody disease prevention food of the present invention is not particularly limited as long as it is caused by a thrombus, and examples thereof include a deep vein thrombosis, a portal vein thrombosis, and a renal static. Venous thrombosis in a group consisting of thrombosis of jugular vein thrombosis, Budd-Chiari syndrome, sacral-clavicular venous thrombosis, cerebral venous sinus thrombosis, and pulmonary thromboembolism, or from cerebral infarction, myocardial infarction, and intestine Interstitial arterial thrombosis: Arterial thrombosis in a group of lower extremity acute arterial thrombosis, hepatic arterial thrombosis, renal artery thrombosis, splenic artery thrombosis, and arteriosclerosis obliterans. The thrombotic disease prevention food of the present invention may be an active ingredient prepared in the above manner and an excipient conventionally used in the food field (for example, starch or paste f, cellulose, lactose, maltose, reduced lactose, reduced maltose, sorbus) Sugar alcohol, #露醇醇, erythritol, xylitol, etc.) or adjuvants (eg 'solvents, dispersion media, coatings, stabilizers, diluents, preservation 1 antiseptics|, bactericides, antifungal agents And an osmotic pressure agent, an absorption inhibiting agent 'disintegrant, an emulsifier, a binder, a lubricant, a pigment, etc.) are mixed and made into, for example, tablets, capsules, and granules by a preparation method conventionally used in the food field. 'Powder, extract, solution, slurry, suspension 1612〇3.d〇 (201233339 liquid, emulsion form. The bloody disease prevention food of the present invention may contain ★ for the whole food §· my team y The weight of the relative body is about 0.05~the weight of the weight, and more preferably about 〜85. The weight is about °.1, %, and the mangsheng is further preferably about 5~8 〇 Good for about I 〇~5〇% by weight ϊ Active ingredient. "The resulting liquid component is used as a sputum for other aspects of the invention, and also relates to a method for the prevention of thrombotic disease. From the use of (4) enzymatic hydrolysis, corn-killing powder The obtained saccharide is used as a culture medium wire, and a yeast extract as a source is added thereto, and the substrate is inoculated with Bacillus subtilis AK as a nano-bacterium (registered number: FERM P, After the fermentation and ripening, the use of the components of the liquid component produced by the arrogant; the use of the bacterium for the prevention of food for thrombotic diseases; characterized by the blood test for the prevention of thrombotic diseases The present invention will be described in more detail based on the examples, but the present invention is not limited by the examples. [Example 1] Yellow corn starch, 〇.5 kg at 2_3 kg Add 5 kg of purified water to soy peptone, 〇5 kg of rice bran juice, 80 g of calcium chloride, 150 g of salt, heat and dissolve. Then cool it and add 5 g of the temple powder. Enzyme After saccharification, add 1.5 kg of sugar, 1.7 kg of glucose, 450 g of yeast extract (Sakamoto Pharmaceutical Co., Ltd.), 1.5 kg of rice bran, 80 g of sodium sulphate 5 kg of vegetable crush 161203.doc 201233339 juice (cabbage, carrot, axe,,, 篆 早 早 ) ) )) and purified water, so that the total weight is 150 kg. Also, add sodium hydroxide and PH The value was adjusted to be in the range of 7.2 to 7.6. The cells were placed in a culture tank and autoclaved for 20 minutes in 12 passages. After cooling, the Bacillus subtilis Α κ strain was chilled, and in a temperature chamber of C, It is fermented for 6 () days at a pH of 4.5 to 6.5, and then aged at a temperature of 15 ± 2 C & /m to a medium value of 4" to 6 〇 to make it mature for 12 days. . The supernatant was sterilized in 2Q minutes, and allowed to naturally cool to culture (4). After the (4) was filtered with a filter paper, the pH was adjusted to 3.5_37 by citric acid, and further sterilized at 95 ° t, and then placed in a 5 gallon tank at a temperature of 90 ° C or higher. In the above manner, 125 liters of a liquid food original solution (enm) was obtained. The general analysis results in 100 g of the obtained food stock solution (ENM) are shown in Table 1 below. Further, as for the above-mentioned food stock solution, a column (TSKgel G2500PWXL) manufactured by T〇s〇h Co., Ltd. was used, and a liquid high-speed layer in which the mobile phase was a mixture of water, acetonitrile and trifluoroacetic acid of 55:45 ··0.1 was used. The size exclusion chromatography (SEC) was determined by analyzing the image (manufactured by Sh〇dex Co., Ltd.: GPC SYSTEM-21), and the detector sensitivity (ultraviolet spectrophotometer: mV) and the standard product having a known molecular weight were used. The dissolution time was compared. The molecular weight distribution of the analysis is shown in Fig. 1, and the ratio (percentage) of the area of the molecular weight fraction in the figure to the whole is shown in Table 1. [Table η Molecular weight range Peak area percentage (%) 161203.doc 12 201233339 10,000 or more traces 3,000-10,000 2 1,000-3,000 9 500-1,000 9 500 or less 80 total 100 Then, the obtained food liquid (ENM) is contained Whether the low molecular substance is modified or decomposed by heat is evaluated. The molecular weight distribution of the ENM heated at 121 ° C for 30 minutes was measured by size exclusion chromatography using a TSK gel G2500 PWXL column (manufactured by Tosoh Co., Ltd.). Also, the molecular weight distribution of the same batch of ENM (Control ENM) not placed in the high temperature was measured in the same manner. The results of these are shown in Table 2. [Table 2] Molecular weight range 10,000 or more 3.000- 10,000 1.000- 3,000 500-1,000 500 or less
峰面積百分率（％) 熱處理ENM 0 2 8 9 81 100Peak area percentage (%) Heat treatment ENM 0 2 8 9 81 100
峰面積百分率（％) 對照ENM 0 2 8 9 81 100 161203.doc -13- 201233339 分子量分佈測定之結果，ENM即便置於高溫條件下，亦 未發現於低分子組成之分子量分佈中發生變化，因此判明 ENM對熱具有耐性。 又，於所獲得之食品用原液（ENM)中含有之低分子物質 被附上強酸條件之情形時，對是否產生改質或分解進行評 價。 一面保持ENM(pH值3.7)為37°C，一面進行攪拌，加入 鹽酸使pH值為1.2。放置1 5分鐘後，利用氫氧化鈉還原為 原來之pH值3.7，藉由使用TSK gel G2500PWXL管柱 (Tosoh股份有限公司製造）之尺寸排除層析法測定該試樣之 分子量分佈。並且，以同樣之方式對未置於強酸中之同批 次的ENM(對照ENM)之分子量分佈進行測定。將該等之結 果示於表3。 [表3] 分子量範圍 10,000以上 3.000- 10,000 1.000- 3,000 500-1,000 500以下 合計 峰面積百分率（％) 峰面積百分率（％)Peak area percentage (%) Control ENM 0 2 8 9 81 100 161203.doc -13- 201233339 As a result of the molecular weight distribution measurement, ENM was not found to change in the molecular weight distribution of the low molecular composition even under high temperature conditions, It was found that ENM is resistant to heat. Further, in the case where the low molecular substance contained in the obtained stock solution for food (ENM) is attached with a strong acid condition, whether or not the modification or decomposition is evaluated is evaluated. While maintaining the ENM (pH 3.7) at 37 ° C, stirring was carried out, and hydrochloric acid was added to adjust the pH to 1.2. After standing for 15 minutes, it was reduced to the original pH of 3.7 by using sodium hydroxide, and the molecular weight distribution of the sample was measured by size exclusion chromatography using a TSK gel G2500PWXL column (manufactured by Tosoh Co., Ltd.). Further, the molecular weight distribution of the same batch of ENM (Control ENM) not placed in a strong acid was measured in the same manner. The results of these are shown in Table 3. [Table 3] Molecular weight range 10,000 or more 3.000- 10,000 1.000- 3,000 500-1,000 500 or less Total peak area percentage (%) Peak area percentage (%)
強酸處理ENM 對照ENM 0 微量 4 3 10 10 9 9 77 78 100 100 分子量分佈測定之結果，ENM即便置於強酸條件下，亦 161203.doc 14 201233339 幾乎未發現於低分子組成之分子量分佈中發生變化，因此 表不ENM亦對強酸具有耐性，於口服接種時亦對具 有耐性。 根據以上之結果’證明丽係對高溫或強酸性條件穩定 之原料，且判明即便於食品加工或口服攝取之情形時’亦 不受高溫或酸性條件之影響，而可進行各種加工處理或利 用。 繼而，將製備完成之ENM裝入濃縮裝置中，於4〇〇c以下 之恤度下，施以真空度2〇·6〇 cmHg之一次濃縮，並於9〇。匸 下進行減菌30分鐘，再次裝入濃縮裝置中，於4〇。〇以下之 /服度下，施以真空度2〇_6〇 cmHg之二次濃縮，獲得濃縮比 調整為20倍之ENM-HL。 使用合成基質S-225 1之ENM-HL之影響纖維蛋白溶酶活 性之效果的研究 繼而’本發明者等人為了直接研究ENM-HL(EnZamin研 究所有限公司製造）之對組織纖維蛋白溶酶原活化因子 (tPA)之纖維蛋白溶酶活性的效果，而使用具有纖維蛋白溶 酶之特異性切割部位的合成基質s_225 1進行研究》利用生 理鹽水階段性稀釋（1〇〇〜〇.丨3容量％)ENM-HL，從而用作樣 品。 於10 μΐ之該樣品中加入90 μΐ之tPA(10 IU/ml)、20 μΐ之 Glu-plasminogen(200 pg/ml)及 100 μΐ之 S-2251(l mM)而進 行反應’母隔2.5分鐘測定450 nm之吸光度2小時，算出其 增加度（ΔΑ450 nm)，從而評價tPA活性。 161203.doc 15 201233339 其結果，發現利用ENM-HL之tPA活性之顯著的亢進（圖 2)。 [實施例2] 使用血纖維蛋白平板之ENM-HL之影響tpA活性之效果的 研究 繼而，本發明者等人使用血纖維蛋白平板法，進行 ENM-HL之對tPA活性之效果的研究》利用生理鹽水階段性 稀釋（100-0.35容量％)ENM-HL，從而用作樣品。將於1〇 μ1 之樣品中加入10 μΐ之tPA(25 IU/ml)者添加於血纖維蛋白培 養jhl中，使其反應24小時，從而對tPA活性進行評價。 其結果’即便於血纖維蛋白平板法中，亦確認利用 ENM-HL之tPA活性亢進效果（圖3)。 [實施例3] ENM-HL·之影響血管内皮細胞中之tpA之活性及產生的效 果之研究 繼而，本發明者等人進行ENM-HL之影響血管内皮細胞 中之tPA活性及tPA產生能力之效果的研究。使用源自小鼠 腦血管内皮之細胞bEnd.3細胞作為血管内皮細胞。於24孔 板上，以2χ 105 ceii/well播種bEnd.3細胞，而實現融合，並 於2天後，以0.001〜〇.1容量％之濃度於無血清培養基中添 加ENM-HL ’進行6小時、12小時、24小時培養。利用 fibrin zymography對培養液中之tPA活性進行評價。 又’自培養24小時後之bEnd.3細胞中萃取RNA(ribonucleic acid ’ 核糖核酸）而合成 cDNA(complementary Deoxyribonucleic •16- 161203.doc 201233339 acid，互補去氧核糖核酸），利用即時PCR(polymerase chain reaction，聚合酶鍵反應）法研究tPA mRNA之表現 量。測定作為内因性對照之GAPDH mRNA之表現量，並 進行修正。 發現藉由添加ENM-HL，而使培養上清中之tPA活性於培 養6小時後、12小時後、24小時後亢進（圖4 A)。又，未發 現由於ENM-HL而使tPA mRNA之表現量增加，因此暗示對 血管内皮細胞之tPA產生未造成影響（圖4B)。 [實施例4] 使用有機體分子間相互作用解析裝置（IASYS)之ENM-HL 之對tPA之結合性的研究 本發明者等人考慮於ENM-HL中含有直接作用於tPA之物 質，而使用有機體分子間相互作用解析裝置（IASYS)，進 行ENM-HL之對tPA之結合性的研究。於比色管中使tPA固 相化，加入ENM-HL(1〜20容量％)，從而對其結合性進行評 價。 其結果，發現ENM-HL之濃度依賴性之對tPA的較強結合 性（圖5)。根據該情況而暗示於ENM-HL中含有較強結合於 tPA之物質。 [實施例5] 小鼠之血液中之ENM-HL之對tPA活性之效果的研究 繼而，本發明者等人為了研究有機體内中之ENM-HL之 效果，而使用C57BL6/J小鼠進行研究。將0.3 ml之ENM-HL(0.008〜25容量％) 口服投予至雄性C57BL6/J小鼠（12週 16l203.doc 17 201233339 ，重25±3 g、各組3隻），於2小時後進行採血，採取血 漿並單離優球蛋白成分。利用Hbrin zymography法對該成 分之tPA活性進行評價。 其結果，於ENM-HL 1容量％、（^丨容量。/。之濃度時發現 tPA活性之亢進（圖6)。 小鼠之血液中之ENM_HL之對tpA活性之持續效果的研究 繼而，為了研究有機體内之tPA活性亢進效果之持續效 果’而將發現tPA活性亢進之1%濃度之ENM-HL 口服投予 至C57BL6/J小鼠’於1、2、3、4小時後進行採血，單離優 球蛋白成分，從而利用fibrin zym〇graphy法對活性進行 評價。 其結果’發現tPA活性之亢進效果於2小時後為最高值， 且保持4小時以上（圖7)。 根據如以上之實驗結果，暗示ENM-HL使血液中之tPA活 性宄進’並且相對長時間保持其效果。 [實施例6] 小鼠之ik液中之培養基添加成分之對tpA活性的效果之 研究 (1)乳酸菌醱酵萃取物 繼而本發明者等人進行有機體内之其他菌醱酵萃取物之 效果的研究。將利用生理鹽水稀釋之乳酸菌醱酵萃取物 (原液’ 1〜50容量％)3〇〇 μΐ 口服投予至雄性C57BL6/J小鼠 (12週齡），自於2小時後採取之血漿中單離優球蛋白成分。 利用 fibrin zymography 法評價該成分之 tPA 及 uPA(urokinase- I6l203.doc 201233339 type plasminogen，尿激酶型纖溶酶原激活物）之活性。 其結果’於乳酸菌醱酵萃取物中，未發現增強tpA及uPA 之活性的效果（圖8)。 (2)納豆菌醱酵萃取物 繼而，本發明者等人進行有機體内之其他培養基添加成 分之效果的研究。將利用生理鹽水稀釋之鈉豆菌醱酵萃取 物（2〇倍濃縮液，〇.2〜25容量％)300 μ1 口服投予至雄性 C5"7BL6/J小鼠（丨2週齡）’自於2小時後採取之血漿中單離 優球蛋白成分。利用fibrin zymograph)^^評價該成分之tpA 及uPA之活性。 其結果，於納豆菌醱酵萃取物中未發現增強tpA及uPA之 活性的效果（圖9)。 [產業上之可利用性] 叙⑽為tPA亦用作腦梗塞等血栓性疾病之治療藥，且 活ί·生几進（活性增強）對該等血栓性疾病之治療及預防有 效:本次明確源自納豆菌之作為低分子肽之成分於有機體 内增強tPA之活性。該成分已製成保健營養食品，因此期 待藉由以食品攝取，而預防血栓性疾病。 本發明之血检性疾病預防食品係屬於營養功能食品、營 養辅助食品、健康輔助食品、特定保健用食品、特別是健 食扣之領域者。本發明之血栓性疾病預防食品係將 削用於食品等並且安全性得以確認之微生物培養物作 為有效成分者，且可簡單地攝取。本發明 防食品可藉由簡單且曰常性地攝取，而有效預防血= 161203.doc 19 201233339 病之發病，並減少與血栓性疾病相關之巨大的醫療費。 【圖式簡單說明】 圖1係表示本發明之有效成分之分子量分佈之圖表。 圖2係表示使用具有纖維蛋白溶酶特異性切割部位之合 成基質S-225 1測定的利用本發明之有效成分之tpA活性的 亢進效果之圖表。 圖3係表示使用血纖維蛋白平板測定之利用本發明之有 效成分之tPA活性的亢進效果之照片及圖表（B)。 圖4係表示使用源自小鼠腦血管内皮之細胞沾以」測定 之培養液中之tPA活性的照片（A)及細胞内tPA之基因表現 量之圓表（B)。 圖5係表示使用有機體分子間相互作用解析裝置（iaSyS) 測定之本發明之有效成分對tPA的鍵結性之圖表。 圖6係表示使小鼠攝取本發明之有效成分，於其2小時後 進行採血之血液中之tPA活性的照片與圖表。 圖7係表示於使小鼠攝取本發明之有效成分的情形時之 小鼠血液中之tPA活性的亢進效果之照片及圖表。 圖8係表示使小鼠攝取乳酸菌醱酵萃取物，於其2小時後 進行採血之企液中之tPA及uPA活性的亢進效果之照片。 圖9係表示使小鼠攝取納豆菌醱酵萃取物，於其2小時後 進订採企之众液中之tPA及uPA活性的亢進效果之照片。 16i203.doc -20-Strong acid treatment ENM Control ENM 0 Trace 4 3 10 10 9 9 77 78 100 100 As a result of molecular weight distribution measurement, ENM is not found in the molecular weight distribution of low molecular composition even when placed under strong acid conditions, 161203.doc 14 201233339 Therefore, it is not resistant to strong acid, and it is also resistant to oral inoculation. According to the above results, the raw materials which are stable to high temperature or strong acidic conditions are confirmed, and it is found that even in the case of food processing or oral ingestion, it is not affected by high temperature or acidic conditions, and various processing treatments or uses can be performed. Then, the prepared ENM was placed in a concentrating device, and concentrated at a vacuum of 2 〇·6 〇 cmHg at a temperature of 4 〇〇c or less, and then concentrated at 9 Torr. The bacteria were sterilized for 30 minutes and refilled into a concentrating device at 4 Torr. Under the following conditions, a second degree of vacuum of 2〇_6〇 cmHg was applied to obtain a concentration ratio of 20 times that of ENM-HL. The effect of the influence of the ENM-HL of the synthetic substrate S-225 1 on the activity of the plasmin activity was followed by the inventors of the present invention to directly study the tissue plasmin of ENM-HL (manufactured by EnZamin Research Co., Ltd.). The effect of plasmin activity of the original activating factor (tPA) was studied using a synthetic substrate s_225 1 with a specific cleavage site for plasmin. Phased dilution with physiological saline (1〇〇~〇.丨3) Capacity %) ENM-HL for use as a sample. The reaction was carried out by adding 90 μL of tPA (10 IU/ml), 20 μL of Glu-plasminogen (200 pg/ml) and 100 μM of S-2251 (1 mM) to 10 μΐ of the sample. The absorbance at 450 nm was measured for 2 hours, and the degree of increase (ΔΑ450 nm) was calculated to evaluate the tPA activity. 161203.doc 15 201233339 As a result, significant increase in tPA activity using ENM-HL was found (Fig. 2). [Example 2] Study of the effect of ENM-HL on the use of fibrin plate on the effect of tpA activity, the inventors of the present invention used the fibrin plate method to carry out the study of the effect of ENM-HL on tPA activity. The saline was serially diluted (100-0.35 vol%) ENM-HL to be used as a sample. The tPA activity was evaluated by adding 10 μM of tPA (25 IU/ml) to a sample of 1 μ μl and adding it to fibrin culture jhl for 24 hours. As a result, even in the fibrin plate method, the effect of the tPA activity of ENM-HL was confirmed (Fig. 3). [Example 3] Effect of ENM-HL· on the activity and production of tpA in vascular endothelial cells The inventors of the present invention performed the influence of ENM-HL on tPA activity and tPA production ability in vascular endothelial cells. Research on effects. Bend.3 cells derived from mouse vascular endothelium were used as vascular endothelial cells. The bEnd.3 cells were seeded at 2χ 105 ceii/well on a 24-well plate to achieve fusion, and after 2 days, ENM-HL ' was added to the serum-free medium at a concentration of 0.001 to 0.1% by volume. Cultured in hours, 12 hours, and 24 hours. The tPA activity in the culture broth was evaluated by fibrin zymography. In addition, RNA was extracted from bEnd.3 cells cultured 24 hours later (ribonucleic acid 'ribonucleic acid) to synthesize cDNA (complementary Deoxyribonucleic • 16-161203.doc 201233339 acid, complementary deoxyribonucleic acid), using real-time PCR (polymerase chain) Reaction, polymerase bond reaction) method to study the expression of tPA mRNA. The amount of expression of GAPDH mRNA as an endogenous control was measured and corrected. It was found that the tPA activity in the culture supernatant was increased by 6 hours, 12 hours, and 24 hours after the addition of ENM-HL (Fig. 4A). Further, it was not found that the expression amount of tPA mRNA was increased by ENM-HL, and thus it was suggested that there was no effect on the tPA production of vascular endothelial cells (Fig. 4B). [Example 4] Study on the binding of ENM-HL to tPA using the organic intermolecular interaction analysis device (IASYS) The inventors of the present invention considered that the substance which acts directly on tPA in ENM-HL is used, and the organism is used. The intermolecular interaction analysis device (IASYS) was used to study the binding of ENM-HL to tPA. The tPA was solidified in a colorimetric tube, and ENM-HL (1 to 20% by volume) was added to evaluate the binding property. As a result, the concentration-dependent binding of ENM-HL to tPA was found to be strong (Fig. 5). According to this situation, it is suggested that ENM-HL contains a substance which strongly binds to tPA. [Example 5] Study on the effect of ENM-HL in the blood of mice on the activity of tPA. Then, the inventors of the present invention conducted research using C57BL6/J mice in order to study the effect of ENM-HL in the organism. . 0.3 ml of ENM-HL (0.008~25 vol%) was orally administered to male C57BL6/J mice (12 weeks 16l203.doc 17 201233339, weight 25±3 g, 3 groups), and was performed 2 hours later. Blood was collected, and plasma was taken and separated from the euglobulin component. The tPA activity of the fraction was evaluated by Hbrin zymography. As a result, it was found that the activity of tPA activity was increased at the concentration of ENM-HL 1 and % (Fig. 6). The effect of ENM_HL in the blood of mice on the sustained effect of tpA activity was followed by To study the sustained effect of the intensive effect of tPA activity in the organism', and to find the 1% concentration of ENM-HL of the tPA activity to be administered orally to C57BL6/J mice for blood collection after 1, 2, 3, and 4 hours. The activity of the euglobulin component was evaluated by the fibrin zym〇graphy method. The results showed that the hyperactivity of tPA activity was the highest after 2 hours and remained for more than 4 hours (Fig. 7). As a result, it was suggested that ENM-HL promoted the tPA activity in the blood and maintained its effect for a relatively long period of time. [Example 6] Study on the effect of the medium addition component in the ik solution of mice on tpA activity (1) Lactic acid bacteria The study of the effect of the other extracts of the bacterium by the inventors of the present invention, etc. The lactic acid bacteria extract extract (stock solution '1 to 50% by volume) 3〇〇μΐ orally diluted with physiological saline Invested to C57BL6/J mice (12 weeks old), plasma mononuclear globulin components taken after 2 hours. The tPA and uPA of this component were evaluated by fibrin zymography (urokinase-I6l203.doc 201233339 type plasminogen, urine) The activity of the kinase-type plasminogen activator). The results showed that the activity of enhancing the activity of tpA and uPA was not observed in the lactic acid bacteria extract (Fig. 8). (2) The natto fermentation extract was followed by The inventors of the present invention conducted an effect of adding a component to another medium in the organic body. Oral administration of sodium natto fermentation extract (2 times concentrated solution, 2 to 25 % by volume) diluted with physiological saline 300 μl was orally administered. To male C5 "7BL6/J mice (丨 2 weeks old)' plasma-isolated euglobulin component taken from 2 hours later. The activity of tpA and uPA of this component was evaluated by fibrin zymograph. As a result, no effect of enhancing the activity of tpA and uPA was found in the natto extract extract (Fig. 9). [Industrial Applicability] (10) is that tPA is also used as a therapeutic drug for thrombotic diseases such as cerebral infarction, and it is effective for the treatment and prevention of such thrombotic diseases: this time It is clear that a component derived from natto as a low molecular peptide enhances the activity of tPA in an organism. This ingredient has been formulated into a nutraceutical food, and therefore it is expected to prevent thrombotic diseases by ingesting food. The blood test disease prevention food of the present invention belongs to the fields of nutritional functional foods, nutritional supplement foods, health supplement foods, specific health foods, and particularly food snacks. The thrombotic disease-preventing food of the present invention is a microbial culture in which foods and the like are confirmed and the safety is confirmed as an active ingredient, and can be easily taken. The food control of the present invention can effectively prevent the onset of blood by simple and regular intake, and reduce the huge medical expenses associated with thrombotic diseases. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a graph showing the molecular weight distribution of the active ingredient of the present invention. Fig. 2 is a graph showing the effect of the hyperactivity of tpA activity using the active ingredient of the present invention measured using a synthetic substrate S-225 1 having a cleavage site specific for plasmin. Fig. 3 is a photograph and a graph (B) showing the effect of the use of the tPA activity of the active ingredient of the present invention measured using a fibrin plate. Fig. 4 is a photograph (A) showing the tPA activity in the culture solution measured by using the cells derived from the mouse cerebral vascular endothelium, and a round table (B) showing the gene expression amount of the intracellular tPA. Fig. 5 is a graph showing the binding property of the active ingredient of the present invention to tPA measured using an organic intermolecular interaction analysis apparatus (iaSyS). Fig. 6 is a photograph and graph showing the tPA activity in the blood of the blood collection after the mice were ingested with the active ingredient of the present invention 2 hours later. Fig. 7 is a photograph and graph showing the effect of the hyperactivity of tPA activity in the blood of mice when the mouse is ingested with the active ingredient of the present invention. Fig. 8 is a photograph showing the effect of the ingestion of tPA and uPA activity in a blood-collecting liquid obtained by ingesting a lactic acid bacteria extract in a mouse. Fig. 9 is a photograph showing the effect of the ingestion of tPA and uPA activity in the liquid of the natto-fermented extract of the mouse after the mouse was ingested for 2 hours. 16i203.doc -20-