Bacillus amyloliquefaciens V656係株分離自台灣中部土壤之蛋白酶生產菌。本研究以B. amyloliquefaciens V656發酵全脂奶粉生產蛋白酶。B. amyloliquefaciens V656生產蛋白酶之較適培養條件為0.5% milk powder，0.05% MgSO4．7H2O，0.1% K2HPO4 ,於pH 7，100mL，30℃,所得蛋白酶活性為55.15 U/mL。B. amyloliquefaciens V656發酵全脂奶粉所得上清液經硫酸銨沉澱、DEAE-Sepharos陰離子交換層析法及Sephacryl-S100膠體過濾層析，可純化出一種蛋白酶。此蛋白酶之最適反應pH、最適反應溫度、pH安定性及熱安定性分別為7、50℃、6-9及50℃。所得蛋白酶經SDS-PAGE電泳分析測得分子量為29 KDa，EDTA及馬麥特鐵對其具抑制效果。 A protease producing microorganism, Bacillus amyloliquefaciens V656, capable of utilizing whole milk powder as nutrition source was screened out from the soils in central Taiwan. In this research, whole milk powder was fermented by B. amyloliquefaciens V656 to produce protease. An optimal protease activity55.15 U/mL was obtained with a medium containing 0.5% milk powder，0.05% MgSO4．7H2O，0.1% K2HPO4 at pH 7,100 mL, 30℃.A protease was purified from the culture supernatant with ammonium sulfate precipitation followed by DEAE-Sepharose column chromatogrephy and Sephacryl-S100 gel chromatography. The purified protease was stable within pH 6-9. The optimal temperature and pH for the protease were 50℃ and 7 respectively. The molecular weight of the protease was 29 KDa assayed by SDS-PAGE. EDTA and marmite irons showed inhibitory effect on this protease.
