Characterization of pbpA and pbp2 encoding penicillin-binding proteins located on the downstream of clavulanic acid gene cluster in Streptomyces clavuligerus.
Two genes, pbpA (orf18) and pbp2 (orf19) located on the downstream of clavulanic acid (CA) gene cluster of Streptomyces clavuligerus were cloned into pET-28a(+), and confirmed to encode a family of high molecular-weight penicillin-binding proteins (PBPs). Both genes were amplified from genomic DNA by PCR and expressed in E. coli BL21 (DE3). Hydropathy plots of the proteins revealed a single stretch of hydrophobic amino acids indicating them to be transmembrane proteins. Pbp2 had lower affinity to penicillin G compared to PbpA, and was essential to the cell growth in contrast to PbpA.
Ishida
Kenji
K
Institute of Biomolecule Reconstruction, Sun Moon University, 100, Kalsanri, Tangjeonmyun, 336-708, Asansi, Chungnam, Korea.
Hung
Trinh Viet
TV
Liou
Kwangkyoung
K
Lee
Hei Chan
HC
Shin
Chang-Hun
CH
Sohng
Jae Kyung
JK
D000595:Amino Acid Sequence; D019818:Clavulanic Acid / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D046915:Penicillin-Binding Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
D046915:Penicillin-Binding Proteins; D011994:Recombinant Proteins; D019818:Clavulanic Acid