Purification and Properties of Diethyl p-Phthalate Esterase from Ochrobactrum anthoropi 6-2b

Autor:	Shin-ichiro Suye, Shichao Xu, Chie Komatsu, Ichiro Takahashi
Rok vydání:	2006
Předmět:	Hydrolysis Electrophoresis chemistry.chemical_compound Chromatography chemistry Size-exclusion chromatography Specific activity General Medicine Sodium dodecyl sulfate Polyacrylamide gel electrophoresis Esterase Phenylmethylsulfonyl Fluoride
Zdroj:	Sen'i Gakkaishi. 62:226-231
ISSN:	1884-2259 0037-9875
DOI:	10.2115/fiber.62.226
Popis:	An esterase which hydrolyzes diethyl p-phthalate (diethyl terephthalate, DET) was obtained from Ochrobactrum anthoropi 6-2b. It was purified 19.1 fold to electrophoretic homogeneity with a yield of 26%. The purified esterase had a specific activity of 32.2umol min-1 mg-1-protein based on the hydrolysis of DET at pH 7.0 and 30deg. The enzyme showed maximum activity at pH 8.0 and was stable at pH 6.0-8.0 with optimum stability at pH 6.0. Its optimal temperature in reaction was 50deg., and it was stable below 40deg. Esterase activity was 89% inhibited by adding 1.0 mM Ni2+ and 82% inhibited by 1.0 mM phenylmethylsulfonyl fluoride. The esterase obeyed Michaelis-Menten kinetics, and in the hydrolysis of DET, Vmax and Km were 37.3 umol min-1 mg-1-protein and 1.17 mM, respectively. It was able to hydrolyze a number of p-phthalates and o-phthalates. Its molecular weight was 40,000 by gel filtration chromatography and 20,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis.
Databáze:	OpenAIRE
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