以納豆菌生產維生素K2 (MK-7) 之研究

Production of Vitamin K2 (MK-7) from Bacillus subtilis natto

10.6342/NTU201701623

曾建勳

MK-7 ； 維生素K2 ； 饋料培養 ； 納豆菌 ； 發酵槽 ； MK-7 ； Vitamin K2 ； Fed-batch ； Bacillus subtilis natto ； Bioreactor

臺灣大學生化科技學系學位論文

2017年

碩士

李昆達

繁體中文

維生素K化合物，尤其是維生素K2 (MK-7) ，已被證實可以改善心血管疾病、骨質疏鬆等疾病。多篇研究指出，MK-7的功能性、半衰期、吸收率及體內利用率皆較其它維生素K優異，但在自然界中MK-7含量低，僅在少量發酵食品中可以獲得，例如納豆，是目前世界上MK-7含量最多的食物，而MK-7主要存在於納豆菌 (Bacillus subtilis natto) 的細胞膜上。從本實驗室所收集的20株納豆菌中篩選出具有生產MK-7潛力的納豆菌NTU-19，再利用前人研究之生產MK-7最佳化培養基配方進行改良，經搖瓶培養改良培養基配方，再放大至5公升生物反應器中進行培養。經過培養基最佳化，搖瓶培養120小時後，可生產MK-7濃度36.68 mg/L。在生物反應器放大培養中，本研究採用冷藏方式，發現可以使MK-7濃度提早達到高點，使培養時間縮短將近一半，是未被報導過的培養方式。目前，生物反應器批次培養120小時，MK-7濃度為37.12 mg/L，而饋料培養120小時則可以將產量提高1.5倍，濃度53.89 mg/L。

The Vitamin K series, particularly Vitamin K2 (MK-7), has been confirmed that can reduce both cardiovascular disease and prevent osteoporosis. Dozens of scientific reports have shown that MK-7 has the best bioavailability, half-life, absorption rate, and function in the human body. However, MK-7 can only be found in some fermented foods, such as Natto, which is the traditional fermented soy bean food in Japan. Previous reports show that Natto has the most abundant MK-7 in the world. The MK-7 in the Natto is synthesized by Bacillus subtilis natto. Bacillus subtilis strain NTU-19 which was selected from 20 strains of Bacillus subtilis natto collected in our laboratory. NTU-19 shows the most MK-7 production potential. Then, we adjust the medium for MK-7 production based on previous report; make it more suitable to NTU-19 for MK-7 production. In Hinton flask culture, NTU-19 produced MK-7 concentration to 36.68 mg/L after 120 hours at 37oC, 125 rpm. In 5-L bioreactor batch culture, the NTU-19 produced MK-7 concentration to 37.12 mg/L. Finally, in 5-L bioreactor fed-batch culture, NTU-19 produced MK-7 concentration to 53.89 mg/L after 120 hours, the total MK-7 concentration increase 1.5- fold during fermentation. Moreover, in this study, we found the special cold effect that can reduce almost half time of culture.

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