Bacterial resistance to vancomycin: overproduction, purification, and characterization of VanC2 from Enterococcus casseliflavus as a D-Ala-D-Ser ligase.
Bacterial resistance to vancomycin: overproduction, purification, and characterization of VanC2 from Enterococcus casseliflavus as a D-Ala-D-Ser ligase.
复制标题
细菌对万古霉素的耐药性：来自卡氏肠球菌的 VanC2 作为 D-Ala-D-Ser 连接酶的过量生产、纯化和表征。
DOI：
10.1073/pnas.94.19.10040
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发表时间：
1997
影响因子：
11.1
通讯作者：
Walsh,CT
中科院分区：
文献类型：
--
作者：
Park,IS;Lin,CH;Walsh,CT
肠球菌对万古霉素临床耐药的VanC表型表现为鸡黄肠球菌和casseliflaventerococcus。通过检测细胞前体udp - n -乙酰氨基酸五肽中间终止于ind-Ala-d-Ser而不是d- ala -d- ala，预测VanC连接酶为ad-Ala-d-Ser而不是ad-Ala-d-Ala连接酶。过量生产。casseliflavusATCC 25788vanc2基因在大肠杆菌中，其纯化到均匀性，可以证明atp依赖的ala -d- ser连接酶活性。对c端位置的- ala进行400倍的选择，形成- ala -d- ser /d- ala二肽的kcat/Km2（Km2=Kmford-Ser或c端- ala）比值为270/0.69。VanC2也具有大量的ala -d- asn连接酶活性（kcat/Km2= 74 mM−1min−1）。
The VanC phenotype for clinical resistance of enterococci to vancomycin is exhibited byEnterococcus gallinarumandEnterococcus casseliflavus. Based on the detection of the cell precursor UDP-N-acetylmuramic acid pentapeptide intermediate terminating ind-Ala-d-Ser instead ofd-Ala-d-Ala, it has been predicted that the VanC ligase would be ad-Ala-d-Ser rather than ad-Ala-d-Ala ligase. Overproduction of theE. casseliflavusATCC 25788vanC2gene inEscherichia coliand its purification to homogeneity allowed demonstration of ATP-dependentd-Ala-d-Ser ligase activity. Thekcat/Km2(Km2=Kmford-Ser or C-terminald-Ala) ratio ford-Ala-d-Ser/d-Ala-d-Ala dipeptide formation is 270/0.69 for a 400-fold selection againstd-Ala in the C-terminal position. VanC2 also has substantiald-Ala-d-Asn ligase activity (kcat/Km2= 74 mM−1min−1).