Screening of a Fibrinolytic Enzyme from Microorganisms and its Utilization for Food

研究了担子菌（蘑菇）血液相关生理活性物质的筛选，以期开发新型健康功能性食品。担子菌菌丝在2%麦芽培养基上摇培养2 ~ 3周，温度为251℃。用纤维蛋白平板法测定培养液的纤溶活性，用凝血仪测定凝血酶时间（TT）测定培养液的抗凝血酶活性。结果表明，农、野生侧耳菇和野生侧耳菇均具有较高的纤溶酶活性，其纤溶酶经部分纯化后均为丝氨酸家族蛋白酶。另一方面，在野生的硫磺蒿中发现了较高的抗凝血酶活性，这在人工养殖中是困难的。通过细胞融合，获得了与寄主具有相同乳酸脱氢酶同工酶模式的融合体。尝试利用具有溶纤酶或抗凝血酶活性的担子菌（蘑菇）开发发酵食品。以大米、大麦、果汁、大豆和牛奶为原料的食品的发明，仅靠担子菌就产生了像清酒一样的、像啤酒一样的和像葡萄酒一样的饮料。除酒精饮料外，奶酪、味噌、纳豆、面包等具有纤溶酶或抗凝血酶活性和含有抗癌β-D-gulcan的发酵食品也开始出现。

Screening of blood related physiological active substances was investigated in the purpose of the development of a new healthy and functional food by basidiomycetes(mushroom). The mycelium of basidiomycetes was cultivated for 2-3 weeks at 251 on 2% malt medium under shaking. The culture fluid was assayed for the fibrinolytic activity by fibrin plate method, and for the anti-thrombin acitivity by thrombin time(TT) using blood coagulometer.As the result, a high fibrinolytic activity was found in the agricultural and wild species of Pleurotus cornucopiae, and in the wild species Trichploma sp. The fibrinolytic enzymes from these species were partially purified and characterized as a serine family proteinase both. On the other hand, a high anti-thrombin activity was found in the wild species of Laetiporus sulphureus, which is difficult in artificial cultivation. By cell fusion with Hypsizigus marmoreus, a fusant having same isozyme pattern of lactic acid dehydrogenase with host was obtained.The development of fermented foods using basidiomycetes(mushroom) which have a fibrinolytic activity or anti-thrombin activity was attempted. The invention of foods from rice, barley, fruit juice, soybean and milk as a food material was resulted in sake-like, beer-like and wine-like drinks by basidiomycetes only. In addition to alcohol drinks, the fermented foods of cheese, miso, natto and bread/having fibrinolytic activity or anti-thrombin activity, and or containing anti-cancer β-D-gulcan were originated.