SUBSTITUTION OF ASPARTIC ACID-217 OF CITROBACTER-FREUNDII CEPHALOSPORINASE AND PROPERTIES OF THE MUTANT ENZYMES

被引:16
作者
TSUKAMOTO, K [1 ]
KIKURA, R [1 ]
OHNO, R [1 ]
SAWAI, T [1 ]
机构
[1] CHIBA UNIV,FAC PHARMACEUT SCI,DIV MICROBIAL CHEM,1-33 YAYOI CHO,CHIBA 260,JAPAN
关键词
Active site; Cephalosporinase; Citrobacter freundii; Oxyimino-cephalosporin; Site-directed mutagenesis; β-Lactamase;
D O I
10.1016/0014-5793(90)80250-M
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
On the assumption that Asp-217 of a Citrobacter freundii cephalosporinase forms a salt-bridge with the conserved Lys-67, Asp-217 was changed to glutamic acid, threonine or lysine. The mutant enzymes retained about the same level of activity as that of the wild-type enzyme, and the participation of Asp-217 in the salt-bridge was ruled out. However, the mutations resulted in an increase in hydrolytic activity toward oxyimino-cephalosporins such as cefuroxime, cefmenoxime and ceftazidime, suggesting a possible mechanism of the bacterial resistance to the novel β-lactams by a single mutation in cephalosporinases. © 1990.
引用
收藏
页码:211 / 214
页数:4
相关论文
共 27 条
  • [11] Miller J.H., 1972, EXPT MOL GENETICS, P1
  • [12] MICRO-IODOMETRIC ASSAY FOR PENICILLINASE
    NOVICK, RP
    [J]. BIOCHEMICAL JOURNAL, 1962, 83 (02) : 236 - &
  • [13] CEFUROXIME - NEW CEPHALOSPORIN ANTIBIOTIC
    OCALLAGHAN, CH
    SYKES, RB
    RYAN, DM
    FOORD, RD
    MUGGLETON, PW
    [J]. JOURNAL OF ANTIBIOTICS, 1976, 29 (01) : 29 - 37
  • [14] REFINED CRYSTAL-STRUCTURE OF BETA-LACTAMASE FROM CITROBACTER-FREUNDII INDICATES A MECHANISM FOR BETA-LACTAM HYDROLYSIS
    OEFNER, C
    DARCY, A
    DALY, JJ
    GUBERNATOR, K
    CHARNAS, RL
    HEINZE, I
    HUBSCHWERLEN, C
    WINKLER, FK
    [J]. NATURE, 1990, 343 (6255) : 284 - 288
  • [15] COMPARATIVE-STUDY OF 7 CEPHALOSPORINS - SUSCEPTIBILITY TO BETA-LACTAMASES AND ABILITY TO PENETRATE SURFACE-LAYERS OF ESCHERICHIA-COLI
    RICHMOND, MH
    WOTTON, S
    [J]. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 1976, 10 (02) : 219 - 222
  • [16] DNA SEQUENCING WITH CHAIN-TERMINATING INHIBITORS
    SANGER, F
    NICKLEN, S
    COULSON, AR
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) : 5463 - 5467
  • [17] SAWAI T, 1988, REV INFECT DIS, V10, P721
  • [18] A SET OF BACTERIAL STRAINS FOR EVALUATION OF BETA-LACTAMASE-STABILITY OF BETA-LACTAM ANTIBIOTICS
    SAWAI, T
    YOSHIDA, T
    TSUKAMOTO, K
    YAMAGISHI, S
    [J]. JOURNAL OF ANTIBIOTICS, 1981, 34 (10) : 1318 - 1326
  • [19] SAWAI T, 1982, J BACTERIOL, V152, P567
  • [20] SIROT J, 1988, REV INFECT DIS, V10, P850