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- Title
以Plackett-Burman Design 與反應曲面法改良枯草芽孢桿菌Lo6 蛋白酶 活性.
- Authors
余姿穎; 陳文如; 魏佳俐; 陳國隆
- Abstract
Bacillus subtilis Lo6(Lo6)分泌之蛋白酶已被證實與白肉雞生長性狀有正相關。本研究 旨在探討液態培養下最大化Lo6 蛋白酶活性。實驗先以Plackett-Burman Design(PBD) 挑選出顯著影響酵素活性之因子, 再以Box-Behnken Design 進行反應曲面法最大化蛋 白酶活性。在中性蛋白酶, 顯著影響的變因為培養時間、葡萄糖、CaCl2、溫度、K2HPO4、 轉速、尿素、KH2PO4 和大豆粕(P < 0.05)。顯著影響鹼性蛋白酶的變因為培養時間、 葡萄糖、CaCl2、溫度、K2HPO4、尿素、轉速、NaCl(P < 0.05)。尿素影響係數為負, 金屬離子中只有K2HPO4 影響係數為正。在溫度30℃、培養時間72 小時、葡萄糖0.86% 及K2HPO4 1.0%時中性、鹼性蛋白酶活性為1385 U 及1242 U。進一步探討發現溫度在 26℃時活性較高。綜上所述, Lo6 在溫度26℃下培養72 小時, 大豆粕3%、葡萄糖0.86% 及 K2HPO4 1.0%時具有最高蛋白酶活性. Protease produce by Bacillus subtilis Lo6 (Lo6) has been proved having positive correlation with growth performance of broiler. This study optimize activity Lo6 protease in broth medium. First, using Plackett-Burman Design (PBD) selected the key ingredients, then Box-Behnken Design of response surface methodology (RSM) was applied to optimize the best activity. The most significant variables for neutral protease activity are identified as incubation time, glucose, CaCl2, incubation temperature, K2HPO4, shaker speed, urea, KH2PO4 and soybean meal (P < 0.05). In alkaline protease, the most significant variables are incubation time, glucose, CaCl2, incubation temperature, K2HPO4, urea, shaker speed, NaCl, soybean meal and KH2PO4 (P < 0.05). Urea has a negative effect, and only K2HPO4 has positive effect among metal ions. Using an optimized variables (incubation temperature 30°C, incubation time 72h, glucose 0.86% and K2HPO4 1.0%), neutral and alkaline protease activity increased to 1385 U and 1242 U. Protease activity is higher when incubation temperature is 26°C. To sum up, Lo6 has maximum protease activity at incubation temperature 26°C incubation time 72h, 3% soybean meal, 0.86% glucose and 1.0% K2HPO4.
- Subjects
PROTEOLYTIC enzymes; BACILLUS subtilis; RESPONSE surfaces (Statistics); NEUTRAL proteinases; SOYBEAN meal
- Publication
Journal of the Chinese Society of Animal Science, 2020, Vol 49, p131
- ISSN
0253-9187
- Publication type
Article