Imidase catalyzed hydrolysis of p-D,L-hydroxyphenyl hydantoin

• Main Authors: Pu, Li-Ping, 浦莉平
• Other Authors: Ho Chester S., Yang Yuh-Shyong
• Format: Others
• Language: zh-TW
• Published: 1997
• Online Access: http://ndltd.ncl.edu.tw/handle/49244204321111094108

碩士 === 國立交通大學 === 生物科技研究所 === 85 === Imidase is an industrially important enzyme leading to the productionof D-4-hydroxyphenylglycine(DHPG). DHPG in turn is the key intermediate for the manufacture of important antibiotics such as cefoperazone, cefpiramide, amoxicillin, aspoxicillin and cefadroxil. To be able to produce DHPG, one needs an enzyme that can selectively hydrolyze D-4- hydroxyphenyl hydantoin from its racemic mixure. This enzyme is imidase, also known as D-hydantoinase, dihydropyrimidinase, dihydropyrimidine hydrase, and dihydropyrimidine amidohydrolase. In the past, imidase have been purified from Agrobacterium sp., Pseudomonas striata, beef liversand rat livers. While Taiwan is an major pig exporting country, we trust successful industrial applications of the imidase from pig livers will provide a tremendous value to our agriculture sector. In this thesis, a methodology of purifying imidase from pig livers hasbeen developed. It has been shown that, indeed, the pig liver imidase can selectivelycatalyze p-D,L-hydrophenyl hydantoin which in turn leads to the production of D- hydrophenylglycine(DHPG). The optimum reaction condition for the pig liverimidase appears to be at pH 8.5 and 60 C. The obervation is well in line withthe results shown in the literature. This research has provided important technical insights towards successful bioreactor design and scale-up for theproduction of a wide spectrum of antibiotics.