Book ChapterDOI
Cell-Free Expression of the Heterodimeric Protein Penicillin G Amidase in a Functionally Active Form
H. Bochtler-Hock,Frank Wedekind +1 more
- pp 181-187
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TLDR
Penicillin G amidase from E. coli is a hydrolytic enzyme belonging to the structural superfamily termed N-terminal nucleophile (Ntm) amidohydrolase, which consists of two different subunits-alpha subunit and beta subunit-which are held together by non covalent forces.Abstract:
Penicillin G amidase (PGA, EC 3.5.1.11) from E. coli is a hydrolytic enzyme belonging to the structural superfamily termed N-terminal nucleophile (Ntm) amidohydrolase. It consists of two different subunits-alpha subunit (109 aa, 24 kDa) and beta subunit (557 aa, 62 kDa)-which are held together by non covalent forces. It does not contain any cystein or disulfide bridges. The N-terminal serine residue (Ser290) of the beta subunit constitutes the base in the catalytic cycle and acts as a nucleophile, whereas the alpha subunit mediates the substrate specificity. PGA displays a compact cone-shaped form with a single Cat2+ binding site. Cat2+-binding is presumed to be an important trigger of processing [1].read more
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Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
Penicillin acylase from E. coli: unique gene-protein relation
TL;DR: The nucleotide sequence of the gene (pac) coding for penicillin G acylase from E. coli ATCC 11105 was determined and correlated with the primary structure of the two constituent subunits of this enzyme.
Journal ArticleDOI
Structure of a slow processing precursor penicillin acylase from Escherichia coli reveals the linker peptide blocking the active-site cleft.
Lorraine Hewitt,Volker Kasche,K. Lummer,Richard J. Lewis,Garib N. Murshudov,Chandra S. Verma,Guy Dodson,Keith S. Wilson +7 more
TL;DR: The crystal structure of a slowly processing precursor mutant (Thr263Gly) of penicillin G acylase from Escherichia coli is determined, which reveals that the spacer peptide blocks the entrance to the active-site cleft consistent with an autocatalytic mechanism of maturation.
Journal ArticleDOI
Unusual signal peptide directs penicillin amidase from Escherichia coli to the Tat translocation machinery.
TL;DR: Mutational studies have shown that the hydrophobic core region acts in synergism with the positive charged N-terminal part of the signal peptide as a Tat recognition signal and contributes to the efficient Tat targeting of the pre-pro-penicillin amidase.
Journal ArticleDOI
Effects of site-directed mutations on processing and activities of penicillin G acylase from Escherichia coli ATCC 11105.
TL;DR: Recombination analysis of several mutants demonstrated that the small subunit can be processed only when the large subunit is processed first, and showed that the prerequisite for penicillin G acylase activity is the efficient processing of the largeSubunit and that the maturation of the smallSubunit does not affect the enzymatic activity.