Bioactive function of alkaline protease inhibitor in soy bean miso.

大豆味噌中碱性蛋白酶抑制剂的生物活性作用。

• 批准号: 61580056
• 负责人: ITO Kiyoe
• 金额: $ 1.34万
• 依托单位: Faculty of Education, Tokyo Gakugei University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61580056/
• 关键词: bioactive function; antimicrobial test; alkaline protease inhibitor; inhibition of cleavage of fertilized egg; soy bean miso; rice koji; イトマキヒトデ; アルカリ性; プロテアーゼ; インヒビター

We tried to isolate alkaline protease inhibitor(API) and studied its physicochemical properties and bioactive function. The API activity was detected in two kinds of soy bean misos(Edo-miso and Saikyo-miso) and rice koji which is the raw material for processing soy bean miso. The inhibiting activity to AP was strong in the order of rice koji, Saikyo-miso and Edo-miso, when tested by using crude preparations of API. The API was further purified from rice koji by Sephadex G-50 gel filtration and DEAE-Toyopearl 650S ion exchenge column chromatography. The molecular weight of the purified API was about 20,000 and the pI was 5.6. The API activity was scarcely affected by heated at 70゜C for 30 min. Even heated at 100゜C for 20 min, 76% of the API activity was detected, so the API was high in heatstability. It showed the optimum pH at around 9 and was stable between pH9 and 11.Antimicrobial test was carried out by auto-biography using 8 species of microorganisms, but API did not suppress the growth of these species. But, API inhibits the cleavage of 32 to 64 stage fertilized egg of starfish(Asterina pectinifera) and the fact suggested the inhibitory function of API in DNA synthesis. From the result, API in miso may have some bioactive functions to human body such as suppressing the growth of eukaryotic organisms.

我们尝试分离碱性蛋白酶抑制物(API)，并对其理化性质和生物活性进行了研究。检测了两种大豆酱油(江户酱油和盛京酱油)和加工大豆酱油的原料米曲中的API活性。用API粗制剂测定，其对AP的抑制活性依次为米曲、Saikyo-miso、Edo-Miso。通过Sephadex G-50凝胶过滤和DEAE-Toyota opearl 650S离子交换柱层析，从米曲中进一步纯化出API。纯化的API相对分子质量约为20,000，等电点为5.6。70゜C加热30min对酶活力影响不大。即使在100゜C下加热20min，也能检测到76%的活性，因此该API具有很高的热稳定性。最适pH在9左右，最适pH在9~11之间比较稳定。对8种微生物进行了抑菌试验，但API对这些微生物的生长没有抑制作用。但是，API对海星(Asterina Putinifera)32-期受精卵的卵裂有抑制作用，提示API在DNA合成中具有抑制作用。由此可见，酱油中的API可能对人体具有一定的生物活性，如抑制真核生物的生长。