Changing glycine 21 for glutamic acid in the β-subunit of penicillin G acylase from Kluyvera citrophila prevents protein maturation

1992 ◽  
Vol 36 (5) ◽  
pp. 659-662 ◽  
Author(s):  
Ignacio Prieto ◽  
María del Carmen Rodríguez ◽  
Gabriel Márquez ◽  
Agustín Pérez-Aranda ◽  
José Luis Barbero
Keyword(s):  
Glutamic Acid ◽  
Penicillin G Acylase ◽  
Penicillin G ◽  
Β Subunit ◽  
Protein Maturation

scholarly journals Processing and functional display of the 86 kDa heterodimeric penicillin G acylase on the surface of phage fd

1999 ◽  
Vol 342 (2) ◽  
pp. 415-422 ◽  
Author(s):  
Raymond M. D. VERHAERT ◽  
Jan VAN DUIN ◽  
Wim J. QUAX
Keyword(s):  
Coat Protein ◽  
Signal Sequence ◽  
Protein A ◽  
Alcaligenes Faecalis ◽  
Penicillin Acylase ◽  
Penicillin G Acylase ◽  
Penicillin G ◽  
Β Subunit ◽  
Extracellular Medium ◽  
Α Subunit

The large heterodimeric penicillin G acylase from Alcaligenes faecalis was displayed on the surface of phage fd. We fused the coding sequence (α subunit-internal peptide-β subunit) to the gene of a phage coat protein. A modified g3p signal sequence was used to direct the polypeptide to the periplasm. Here we show that a heterodimeric enzyme can be expressed as a fusion protein that matures to an active biocatalyst connected to the coat protein of phage fd, resulting in a phage to which the β-subunit is covalently linked and the α-subunit is non-covalently attached. The enzyme can be displayed either fused to the minor coat protein g3p or fused to the major coat protein g8p. In both cases the penicillin G acylase on the phage has the same Michaelis constant as its freely soluble counterpart, indicating a proper folding and catalytic activity of the displayed enzyme. The display of the heterodimer on phage not only allows its further use in protein engineering but also offers the possibility of applying this technology for the excretion of the enzyme into the extracellular medium, facilitating purification of the protein. With the example of penicillin acylase the upper limit for a protein to become functionally displayed by phage fd has been further explored. Polyvalent display was not observed despite the use of genetic constructs designed for this aim. These results are discussed in relation to the pore size being formed by the g4p multimer.

scholarly journals Improvement of Catalytic Properties of Escherichia coli Penicillin G Acylase Immobilized on Glyoxyl Agarose by Addition of a Six-Amino-Acid Tag

2005 ◽  
Vol 71 (12) ◽  
pp. 8937-8940 ◽  
Author(s):  
Francesca Scaramozzino ◽  
Ilona Estruch ◽  
Paola Rossolillo ◽  
Marco Terreni ◽  
Alessandra M. Albertini
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Catalytic Properties ◽  
Penicillin G Acylase ◽  
Penicillin G ◽  
Protein Maturation ◽  
Β Chain

ABSTRACT A tag of three lysines alternating with three glycines was added to the C-terminal end of the β chain of penicillin G acylase (PGA). This modification improved the immobilization efficiency of PGA on glyoxyl agarose and the catalytic properties of the PGA derivative, although it impaired the posttranslational steps of overexpressed protein maturation.

Resolution of α/β-amino acids by enantioselective penicillin G acylase from Achromobacter sp .

2015 ◽  
Vol 122 ◽  
pp. 240-247 ◽  
Author(s):  
Michal Grulich ◽  
Jan Brezovský ◽  
Václav ŠtĿpánek ◽  
Andrea Palyzová ◽  
Eva Kyslíková ◽  
...  
Keyword(s):  
Amino Acids ◽  
Penicillin G Acylase ◽  
Penicillin G ◽  
Achromobacter Sp

Porous thiiranyl polymers: Newer supports for immobilization of penicillin G acylase

1997 ◽  
Vol 33 (9) ◽  
pp. 1481-1485 ◽  
Author(s):  
S. Skaria ◽  
E.Sreenivasa Rao ◽  
S. Ponrathnam ◽  
K.K. Kumar ◽  
J.G. Shewale
Keyword(s):  
Penicillin G Acylase ◽  
Penicillin G

Inoculum Studies in Production of Penicillin G Acylase by Bacillus megaterium ATCC 14945

2002 ◽  
pp. 679-686 ◽  
Author(s):  
Laura M. Pinotti ◽  
Rosineide G. Silva ◽  
Roberto C. Giordano ◽  
Raquel L. C. Giordano
Keyword(s):  
Bacillus Megaterium ◽  
Penicillin G Acylase ◽  
Penicillin G
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