Structure, Processing and Catalytic Action of Penicillin Acylase

Virden, Richard

doi:10.1080/02648725.1990.10647869

Cited by 21 publications

(14 citation statements)

References 101 publications

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“…Then the enzyme becomes active by an autoproteolytic process that releases an internal peptide and divides the protein into two dissimilar subunits (␣ and ␤) (137). Pac activity is consistent with an acyl enzyme mechanism involving the N-terminal serine of the ␤ chain as the active site (327). The three-dimensional structure of the Pac protein is known (80), and the enzyme has been classified within a new Ntn-hydrolase family (308).…”

Section: Penicillin G Acylasementioning

confidence: 99%

“…Moreover, it hydrolyzes esters and amides of other aromatic and aliphatic acids such as thyenylacetic acid and hexanoic acid (327). Therefore, it appears to play a role as a scavenger enzyme for many different natural esters and amides which after their hydrolysis can be transported and mineralized by central catabolic pathways (182).…”

Section: Penicillin G Acylasementioning

confidence: 99%

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Biodegradation of Aromatic Compounds by Escherichia coli

Dı́az

Ferrández

Prieto

et al. 2001

Microbiol Mol Biol Rev

324

235

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Although Escherichia coli has long been recognized as the best-understood living organism, little was known about its abilities to use aromatic compounds as sole carbon and energy sources. This review gives an extensive overview of the current knowledge of the catabolism of aromatic compounds by E. coli. After giving a general overview of the aromatic compounds that E. coli strains encounter and mineralize in the different habitats that they colonize, we provide an up-to-date status report on the genes and proteins involved in the catabolism of such compounds, namely, several aromatic acids (phenylacetic acid, 3- and 4-hydroxyphenylacetic acid, phenylpropionic acid, 3-hydroxyphenylpropionic acid, and 3-hydroxycinnamic acid) and amines (phenylethylamine, tyramine, and dopamine). Other enzymatic activities acting on aromatic compounds in E. coli are also reviewed and evaluated. The review also reflects the present impact of genomic research and how the analysis of the whole E. coli genome reveals novel aromatic catabolic functions. Moreover, evolutionary considerations derived from sequence comparisons between the aromatic catabolic clusters of E. coli and homologous clusters from an increasing number of bacteria are also discussed. The recent progress in the understanding of the fundamentals that govern the degradation of aromatic compounds in E. coli makes this bacterium a very useful model system to decipher biochemical, genetic, evolutionary, and ecological aspects of the catabolism of such compounds. In the last part of the review, we discuss strategies and concepts to metabolically engineer E. coli to suit specific needs for biodegradation and biotransformation of aromatics and we provide several examples based on selected studies. Finally, conclusions derived from this review may serve as a lead for future research and applications

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Section: Penicillin G Acylasementioning

confidence: 99%

Section: Penicillin G Acylasementioning

confidence: 99%

Biodegradation of Aromatic Compounds by Escherichia coli

Dı́az

Ferrández

Prieto

et al. 2001

Microbiol Mol Biol Rev

324

235

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“…Although di¡erent PGAs have been described in many microorganisms, the enzyme from Escherichia coli W ATCC 11105 has been the most extensively studied so far [1]. The expression of the pac gene is subjected to several regulatory controls, including temperature, oxygen, catabolite repression, and induction by phenylacetic acid (PAA) [1,2]. In addition, a complex maturation process is required to render the active form of PGA [2].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, a complex maturation process is required to render the active form of PGA [2]. The enzyme is produced as an inactive cytoplasmic precursor of 93 kDa which be-comes catalytically active after its translocation to the periplasmic space followed by an autoproteolytic disruption into two subunits (K and L) that removes an internal spacer polypeptide [2]. Since the pac gene has been cloned and sequenced [3,4], very few studies have been done on the characterization and regulation of the pac promoter [5^7].…”

Section: Introductionmentioning

confidence: 99%

“…The expression of the pac gene is subjected to several regulatory controls, including temperature, oxygen, catabolite repression, and induction by phenylacetic acid (PAA) [1,2]. In addition, a complex maturation process is required to render the active form of PGA [2]. The enzyme is produced as an inactive cytoplasmic precursor of 93 kDa which be-comes catalytically active after its translocation to the periplasmic space followed by an autoproteolytic disruption into two subunits (K and L) that removes an internal spacer polypeptide [2].…”

Section: Introductionmentioning

confidence: 99%

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New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105

Roa

1999

FEMS Microbiology Letters

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The regulation of the pac gene encoding the penicillin G acylase of Escherichia coli W ATCC 11015 has been investigated by a molecular approach using lacZ as a reporter gene. This analysis revealed that a region of 170 bp located upstream of the pac structural gene contains the regulatory sequences that control its expression. The cAMP receptor protein is involved not only in the catabolite repression of penicillin G acylase production caused by glucose but also in the induction of pac gene expression by phenylacetic acid. Primer extension analyses have demonstrated that the transcription of the pac gene can be initiated from at least three different promoters. Although all these promoters are functional, their relative activity depends on the transcribed gene, the P1 and P3 promoters being more active in the presence of the pac gene, whereas the P2 promoter was stronger when the upstream region of the pac gene was fused to the lacZ reporter. A deletion of the region surrounding the 310 box of the P3 promoter produced a constitutive expression of the fused gene indicating that this sequence is required for phenylacetic acid induction and suggesting that the expression of the pac gene is regulated by a repression mechanism. This work reveals that the regulation of the pac gene is more complex than previously envisioned and provides new clues to investigate further this interesting regulatory system. ß

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Synthesis and secretion of recombinant penicillin G acylase in bacterial L‐forms

Gumpert¹,

Cron

Plapp

et al. 1996

J. Basic Microbiol.

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L‐form strains of Proteus mirabilis and Escherichia coli lacking the cell wall represent an alternative prokaryotic cell system for the production of recombinant proteins (KLESSEN et al. 1988, LAPLACE et al. 1988 a, 1989b). We could demonstrate that they are also able to synthesize the enzyme penicillin G acylase (PAC)1). PAC was processed and secreted into the medium by recombinant L‐form strains. The synthesis of PAC was growth‐associated and stably regulated. Expression, secretion, and processing were not temperature‐dependent and occurred at 26 °C, 32 °C and even 37 °C. The expression vector pHC1 carried the pac gene under the control of the lac UV promoter and a kanamycin resistance gene. It could be maintained in L‐form cells, showing low structural as well as segregational instability. The secretion of the biologically active enzyme into the medium indicated that the postranslational processing of the PAC molecule, including the excision of a 54 amino acid spacer peptide between the α and β subunit, is not carried out in the periplasmic space, but occurs at the cytoplasmic membrane or autocatalytically.

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Structure, Processing and Catalytic Action of Penicillin Acylase

Cited by 21 publications

References 101 publications

Biodegradation of Aromatic Compounds by Escherichia coli

Biodegradation of Aromatic Compounds by Escherichia coli

New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105

Synthesis and secretion of recombinant penicillin G acylase in bacterial L‐forms

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