원문정보
포스터 발표 : 미생물공학
초록
영어
In our previous study, recombinant E.coli harboring flavincontainingmonooxygenase(FMO) gene from Methylophaga aminisulfidivorans MPT produced indigo (853mg/L) and indirubin (≤5mg/L) in tryptophan medium (2g tryptophan, 10g NaCl, 5g yeast extract per liter). The goal of this study is to characterize the FMO and investigate the optimum culture conditions to enhance the amount of indirubin produced. The optimal pH andt emperature for FMO enzyme activity was found to be 8.5 and 45℃, respectively. The enzyme showed high affinity with trimethylamine (TMA). Vmax and Km values for trimethylamin were 19.3 nmol/min/mg protein and 23 μM, respectively. The optimum culture conditions determined by response surface method were as follows: 1g/L tryptophan, 5g/L yeast extract, 3mM cysteine, pH8.0 and temperature at 35℃. Under this condition, 15 times higher amount of indirubin (75.1mg/L) was produced.