Chung, JH Back, JH Lim, JH Park, YI Han, YS 2024-01-21T10:33:23Z 2024-01-21T10:33:23Z 2021-09-01 2002-06-04 0141-0229 https://pubs.kist.re.kr/handle/201004/139441 A hyperthermophilic hydantoinase from Methanococcus jannaschii with an optimum growth at 85degreesC was cloned and expressed in E. coli. The recombinant,hydantoinase was petrified by affinity and anion-exchange chromatography and determined to be homotetrameric protein by gel filtration chromatography. The best substrate for the hydantoinase was D,L-5-hydroxyhydantoin, which has the specific activity of 183.4 U/mg. The optimum pH and temperature for the hydantoinase activity was 8.0 and 80degreesC, respectively. The half-life of the hydantoinase was measured to be 100 min at 90degreesC in the buffer containing 500 mM KCl. Manganese ions were the most effective for the hydantoinase activity. Stereospecificity was determined to be L-specific for the 5-hydroxymethylhydantoin and 5-methylhydantoin by chiral TLC. The activity yields as well as the operational stabilities of the thermostable M. jannaschii hydantoinase could be significantly improved by immobilization method. (C) 2002 Elsevier Science Inc. All rights reserved. English ELSEVIER SCIENCE INC COMPLETE GENOME SEQUENCE L-AMINO-ACIDS MICROBIAL TRANSFORMATION ESCHERICHIA-COLI 5-SUBSTITUTED HYDANTOINS ARTHROBACTER-AURESCENS PURIFICATION EXPRESSION CLONING AMIDOHYDROLASE Thermostable hydantoinase from a hyperthermophilic archaeon, Methanococcus jannaschii Article 10.1016/S0141-0229(02)00047-9 1 ENZYME AND MICROBIAL TECHNOLOGY, v.30, no.7, pp.867 - 874 ENZYME AND MICROBIAL TECHNOLOGY 30 7 867 874 scie scopus 000175961300006 2-s2.0-0037018891 Biotechnology & Applied Microbiology Biotechnology & Applied Microbiology Article COMPLETE GENOME SEQUENCE L-AMINO-ACIDS MICROBIAL TRANSFORMATION ESCHERICHIA-COLI 5-SUBSTITUTED HYDANTOINS ARTHROBACTER-AURESCENS PURIFICATION EXPRESSION CLONING AMIDOHYDROLASE hydantoinase hyperthermophile Methanococcus jannaschii stereospecificity thermostability