Purification and characterization of a novel esterase promising for the production of useful compounds from Microbacterium sp 7-1W

被引:4
作者
Honda, K [1 ]
Kataoka, M [1 ]
Ono, H [1 ]
Sakamoto, K [1 ]
Kita, S [1 ]
Shimizu, S [1 ]
机构
[1] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Sakyo Ku, Kyoto 6068502, Japan
关键词
esterase; serine hydrolase; regioselective hydrolysis; cetraxate; pantothenate; Microbacterium;
D O I
10.1111/j.1574-6968.2002.tb11013.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A novel esterase catalyzing regioselective hydrolysis was purified from the membrane fraction of Microbacterium sp. 7-1W, and characterized. The enzyme was solubilized with Brij 58 and purified 13.8-fold to apparent homogeneity with 2.58% overall recovery. The relative molecular mass of the native enzyme as estimated by gel filtration was more than 600000 Da, and the subunit molecular mass was 62000 Da. The enzyme catalyzed cleavage of the terminal ester bonds of cetraxate esters and pantothenate esters. The K-m and V-max values for methyl cetraxate were 0.380 mM and 7.76 mumole min(-1) mg(-1) protein, respectively. The enzyme was inhibited by serine hydrolase inhibitors. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:221 / 227
页数:7
相关论文
共 19 条
[1]  
Bornscheuer U. T, 1999, HYDROLASES ORGANIC S
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]  
HIRAYAM T, 1988, Patent No. 6328062
[4]  
Junge W., 1984, METHODS ENZYMATIC AN, V4, P2
[5]   A NOVEL NADPH-DEPENDENT CARBONYL REDUCTASE OF CANDIDA-MACEDONIENSIS - PURIFICATION AND CHARACTERIZATION [J].
KATAOKA, M ;
DOI, Y ;
SIM, TS ;
SHIMIZU, S ;
YAMADA, H .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1992, 294 (02) :469-474
[6]   Optical resolution of racemic pantoic acid through microbial stereoselective lactonization in an organic solvent/water two-phase system [J].
Kataoka, M ;
Hirakata, M ;
Sakamoto, K ;
Yamada, H ;
Shimizu, S .
ENZYME AND MICROBIAL TECHNOLOGY, 1996, 19 (04) :307-310
[7]   OPTICAL RESOLUTION OF RACEMIC PANTOLACTONE WITH A NOVEL FUNGAL ENZYME, LACTONOHYDROLASE [J].
KATAOKA, M ;
SHIMIZU, K ;
SAKAMOTO, K ;
YAMADA, H ;
SHIMIZU, S .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1995, 43 (06) :974-977
[8]  
Kataoka M, 1995, APPL MICROBIOL BIOT, V44, P333
[9]   STEREOSPECIFIC REDUCTION OF ETHYL-2'-KETOPANTOTHENATE TO ETHYL D-(+)-PANTOTHENATE WITH MICROBIAL-CELLS AS A CATALYST [J].
KATAOKA, M ;
SHIMIZU, S ;
DOI, Y ;
YAMADA, H .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1990, 56 (11) :3595-3597
[10]   POLYPEPTIDES OF TAIL FIBRES OF BACTERIOPHAGE T4 [J].
KING, J ;
LAEMMLI, UK .
JOURNAL OF MOLECULAR BIOLOGY, 1971, 62 (03) :465-&