Crystallization and preliminary X-ray diffraction analysis of glutaryl-7-aminocephalosporanic acid acylase from Pseudomonas sp GK16

被引:11
作者
Kwon, TH [1 ]
Rhee, S [1 ]
Lee, YS [1 ]
Park, SS [1 ]
Kim, KH [1 ]
机构
[1] Korea Univ, Grad Sch Biotechnol, Seoul 136701, South Korea
关键词
glutaryl-7-aminocephalosporanic acid acylase; protein crystallization; Pseudomonas sp GK16; X-ray diffraction;
D O I
10.1006/jsbi.2000.4256
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Glutaryl-7-aminocephalosporanic acid acylase from Pseudomonas sp. GK16 produces glutaryl-7-aminocephalosporanic acid, a key intermediate for the synthesis of cephem antibiotics. Sequence alignment suggests that the enzyme may belong to the N-terminal nucleophile hydrolase superfamily including penicillin G acylase. The enzyme is an (alpha beta)(2) heterotetramer of two nonidentical subunits. These subunits are derived from a nascent precursor polypeptide that is cleaved proteolytically through a two-step autocatalytic process upon folding. The enzyme has been crystallized using the vapor diffusion method. A bipyramidal crystal form was obtained from a solution containing polyethylene glycol (MW 3350) and calcium chloride. Complete diffraction data sets have been collected up to 2.8 Angstrom resolution. The crystal is tetragonal with the space group P4(1)2(1)2 or P4(3)2(1)2 and the unit cell parameters are a = 5 = 73.5 Angstrom, c 380.3 Angstrom. Considerations of the possible values of V-m account for the presence of a tetramer in the asymmetric unit. (C) 2000 Academic Press.
引用
收藏
页码:79 / 81
页数:3
相关论文
共 15 条
[1]   A PROTEIN CATALYTIC FRAMEWORK WITH AN N-TERMINAL NUCLEOPHILE IS CAPABLE OF SELF-ACTIVATION [J].
BRANNIGAN, JA ;
DODSON, G ;
DUGGLEBY, HJ ;
MOODY, PCE ;
SMITH, JL ;
TOMCHICK, DR ;
MURZIN, AG .
NATURE, 1995, 378 (6555) :416-419
[2]   2-STEP IMMOBILIZED ENZYME CONVERSION OF CEPHALOSPORIN-C TO 7-AMINOCEPHALOSPORANIC ACID [J].
CONLON, HD ;
BAQAI, J ;
BAKER, K ;
SHEN, YQ ;
WONG, BL ;
NOILES, R ;
RAUSCH, CW .
BIOTECHNOLOGY AND BIOENGINEERING, 1995, 46 (06) :510-513
[3]   MODIFICATIONS OF ANTIBIOTICS .2. PREPARATION OF 7-AMINOCEPHALOSPORANIC ACID [J].
FECHTIG, B ;
PETER, H ;
BICKEL, H ;
VISCHER, E .
HELVETICA CHIMICA ACTA, 1968, 51 (05) :1108-&
[4]  
Huber FM., 1972, CEPHALOSPORINS PENIC, P27, DOI 10.1016/B978-0-12-261450-7.50007-1
[5]   7-AMINOCEPHALOSPORANIC ACID PRODUCTION .2. THE ISOLATION AND PROPERTIES OF PSEUDOMONAS-MUTANTS WITH AN ENHANCED PRODUCTIVITY OF 7BETA-(4-CARBOXYBUTANAMIDO)-CEPHALOSPORANIC ACID ACYLASE [J].
ICHIKAWA, S ;
MURAI, Y ;
YAMAMOTO, S ;
SHIBUYA, Y ;
FUJII, T ;
KOMATSU, K ;
KODAIRA, R .
AGRICULTURAL AND BIOLOGICAL CHEMISTRY, 1981, 45 (10) :2225-2229
[6]   STRUCTURE AND EXPRESSION OF CDNA FOR D-AMINO-ACID OXIDASE ACTIVE AGAINST CEPHALOSPORIN-C FROM FUSARIUM-SOLANI [J].
ISOGAI, T ;
ONO, H ;
ISHITANI, Y ;
KOJO, H ;
UEDA, Y ;
KOHSAKA, M .
JOURNAL OF BIOCHEMISTRY, 1990, 108 (06) :1063-1069
[7]   SPARSE-MATRIX SAMPLING - A SCREENING METHOD FOR CRYSTALLIZATION OF PROTEINS [J].
JANCARIK, J ;
KIM, SH .
JOURNAL OF APPLIED CRYSTALLOGRAPHY, 1991, 24 :409-411
[8]   Media processing: A new design target [J].
Lee, RB ;
Smith, MD .
IEEE MICRO, 1996, 16 (04) :6-9
[9]   Two-step autocatalytic processing of the glutaryl 7-aminocephalosporanic acid acylase from Pseudomonas sp. strain GK16 [J].
Lee, YS ;
Park, SS .
JOURNAL OF BACTERIOLOGY, 1998, 180 (17) :4576-4582
[10]   MOLECULAR-CLONING AND STRUCTURE OF THE GENE FOR 7-BETA-(4-CARBOXYBUTANAMIDO)CEPHALOSPORANIC ACID ACYLASE FROM A PSEUDOMONAS STRAIN [J].
MATSUDA, A ;
KOMATSU, KI .
JOURNAL OF BACTERIOLOGY, 1985, 163 (03) :1222-1228