원문정보
초록
영어
Aryl acylamidase (EC 3.5.1.13, AAA) is enzyme catalyzing the hydrolysis of anilides (N-acyl primary aromatic amines) to form an aniline compound and a carboxylic acid anion. AAA has known to be useful for the determination of acylamide drugs (e.g. acetaminophen) as a diagnostic reagent in clinical chemistry. AAA can be used to form an amide bond using a reverse reaction of the enzymes, which is attractive for an enzymatic synthesis of aryl acylamide compounds under a mild condition without byproducts. Here, we present a method to synthesize acetaminophen (paracetamol, TyrenolTM , AA) from aminophenol and acetate by a bacterial aryl acylamidase. For the production of AA, an aryl acylamidase gene from a soil bacterium was cloned and overexpressed in E. coli. In order to optimize the reaction condition for the production of acetaminophen by E. coli expressing AAA, we examined induction
condition, buffer composition, temperature, pH and other factors. The optimum condition was in a
mixture of 100 mM of p-aminophenol and 600 mM of sodium acetate as substrates in 100 mM of sodium phosphate buffer (pH 6.0) at 35 ℃. When 12g wet cell mass/l of E. coli cell induced at 25C in the complex auto-inducing media was used under this condition, 60.5 mM of acetaminophen was produced