不同培养条件对纳豆激酶之生产研究.docVIP

不同培養條件對納豆激酶之生產研究 Nattokinase production by Bacillus subtilis in different culture conditions 研究生：張智琮 Chang, Chih-Tsung 指導教授：謝寶全 Lien, Yi-Yang 【摘要】 本研究由市售納豆產品中篩選出一高活性生產菌株 Bacillus subtilis B171，並分別探討液態及固態醱酵之最佳培養條件，以提高菌株生產活性產量。以 % 麩皮及 % 黃豆為固態醱酵基質，在10 % 接種量，起始水分為65 %，30℃培養溫度下，另添加5 % 0℃進行儲存試驗，發現以5及25℃之儲藏條件下，其納豆激酶活性下降緩慢，經儲存兩個月後，其酵素活性仍可維持80%~90%。 關鍵字：納豆激酶、液態發酵、固態發酵 【Abstract】 A high nattokinase activity producing strain, Bacillus subtilis B17-1, was isolated from commercial natto and used to compare the activity of nattokinase by different fermentation methods in this study. The results showed that 50 mL medium in 250 mL Hinton’s, which contained 2% fructooligosaccharides, 3% defatted soybean, 0.15% CaCO3 and the initial medium pH at 7.0 ± 0.2, shaked with 150 rpm and 1% inoculums size producing the highest nattokinase activity 220 FU/mL at 37℃ cultured for 42 hours. Using 5 Liter fermentor to research the fermentation conditions showed, fermentor contained 2 L medium, 1vvm aeration, stirred speeds with 350 rpm and 1% inoculums, in fermentation process to fed fructooligosaccharides and defatted soybean, producing the highest nattokinase activity 600 FU/mL at 37℃ cultured for 45 hours. The fermentative substrates were wheat bran and soybean in mass ratio of 1:1 in solid state fermentation. Supplement with 5% fructooligosaccharides, 5% defatted soybean and 0.1% CaCO3 enhanced the enzyme productivity considerably. Maximum enzyme activity was 1030 FU/g after fermentation 40 hours at 37℃, initial moisture 65% and 10% inoculums size. Several drying methods were evaluated for the liquid fermentation product, including ultrafiltration, freeze drying and spraying drying. 17200 FU/g of nattokinase was obtained from freeze drying for liquid culture treating with centrifuge. In storage test, the nattokinase powder maintained the residual activity 80-90% that put at 5℃ and 25℃ for 2 month. Keywords：nattokinase, liquid fermentation, solid state fermentation.