Kinetics and mechanism of acyl transfer by penicillin acylases

Konecny, J.; Schneider, A.; Sieber, M.

doi:10.1002/bit.260250212

Cited by 34 publications

(10 citation statements)

References 38 publications

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“…It is based on the use of an activated acyl donor (as ester or amide) to form an acyl-enzyme intermediate that may be further deacylated by the antibiotic nucleus (Fig. 3) to yield the desired antibiotic [4,[49][50][51]. That is, KCS may be carried out only if this acylenzyme is formed, as in the case of all PGA described to date.…”

Section: Kinetically Controlled Synthesis (Kcs)mentioning

confidence: 99%

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic βLactam Antibiotics

Mateo¹,

Abián²,

Grazú³

et al. 2005

MCRO

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The production, under environmentally benign conditions, of efficient and more cost-effective anti-infective agents (available to the whole mankind) is one of most ambitious dreams of the industrial medicinal chemistry.Semi-synthetic β-lactam antibiotics are very effective anti-infective agents. They are very stable and can be used via oral delivery. They exhibit a very wide spectrum of anti-bacterial activity and minimal side effects after being massively used for a very long time. In this way, we can assume that semi-synthetic β-lactam antibiotics are going to be one of the key anti-infective agents in the next years.The condensation of natural or modified antibiotic nuclei with different acyl donor chains is one of the key steps for the industrial synthesis of these anti-infective agents. Up to now, these condensations are mainly carried out through classical chemical methods and this implies a number of economical, ecological and technological drawbacks (high energy requirements, many protection and deprotection steps, utilization of toxic methylene chloride as solvent, etc).Enzyme biocatalysts may be very useful to catalyse these selective condensations under very mild experimental and environmental conditions. In fact, the possibility of using enzymes to carry out such biotransformations, at laboratory scale, has been discussed and demonstrated a long time ago. However, industrial synthesis of β-lactamic antibiotics is still carried out via unfavorable chemical routes. In fact, enzymes have not been produced in the nature to act in industrial reactors; so they are usually very unstable, inhibited by substrates and products and they may not have ideal catalytic properties for industrial uses (high reaction rates, required selectivity, ability to reach quantitative synthetic yields, stable enough to run a number of reaction cycles, etc). These limitations of enzyme biocatalysis become even more significant mainly when the enzyme is going to be used with non-natural substrates, catalysing non-natural processes and working under non-conventional conditions. However, in the last ten years, a great number of papers have reported substantial improvement of these enzymatic synthetic approaches (new enzymes, better enzyme derivatives, improved reaction designs and so on) and it seems that a massive industrial implementation of enzymes in antibiotic synthesis is approaching. In this review, we would like to make a critical discussion of these very interesting advances in the application of enzyme biocatalysts for the industrial synthesis of semi-synthetic antibiotics.

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Section: Kinetically Controlled Synthesis (Kcs)mentioning

confidence: 99%

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic βLactam Antibiotics

Mateo¹,

Abián²,

Grazú³

et al. 2005

MCRO

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“…Although the substrate specificity (Margolin et al 1980) and the kinetic mechanism (Konecny et al 1983) of PA has been extensively investigated, the molecular basis of substrate binding and catalysis, as well as the tridimentional structure of the protein remain to be determined. Nevertheless, it has been established that the catalytic mechanism progresses through an acyl-enzyme intermediate (Konecny et al 1983).…”

Section: Introductionmentioning

confidence: 99%

“…Nevertheless, it has been established that the catalytic mechanism progresses through an acyl-enzyme intermediate (Konecny et al 1983). Using site-directed reagents the existence of an essential serine residue which could be directly involved in the binding of the acyl moiety of the substrate (Siewinski et al 1984) has also been demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Penicillin acylase mutants with altered site-directed activity from Kluyvera citrophila

Priéto¹,

Martín

Arche

et al. 1990

Appl Microbiol Biotechnol

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Oligonucleotide-directed mutagenesis has been used to obtain specific changes in the penicillin acylase gene from Kluyvera citrophila. Wild-type and mutant proteins were purified and the kinetic constants for different substrates were determined. Mutations in Met168 highly decreased the specificity constant of the enzyme for penicillin G, penicillin V and phenylacetyl-4-aminobenzoic acid and the catalytic constant kcat for phenylacetyl-4-aminobenzoic acid. Likewise, the phenylmethylsulphonyl-fluoride sensitivity was significantly decreased. It is concluded that the 168 residue is involved in binding by interaction with the acid moiety of the substrate. A putative penicillin-binding domain was located in penicillin acylase by sequence homology with other penicillin-recognizing enzymes. Lys374 and His481, the conserved amino acid residues that are essential for catalysis in these enzymes, can be changed in penicillin acylase with no changes to the kcat and phenylmethylsulphonyl fluoride reactivity, but change the Km. The likelihood of the existence of this proposed penicillin binding site is discussed. The reported results might be used to alter the substrate specificity of penicillin acylase in order to hydrolyse substrates of industrial significance other than penicillins.

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“…This means that the enzymic reaction mechanism of polymyxin acylase is different from those of usual aminoacylases. Similar enzymes, penicillin acylases, produced from E. COli,4) Bacillus megaterium, 5) or Xanthomonas citri 6 ) are also distinguishable from polymyxin acylase, because they are serine enzymes, inhibited by DFP or PMSF.…”

mentioning

confidence: 98%

Polymyxin Acylase: An Enzyme Causing IntramolecularN²⇄N⁶Acyl Transfer inN-Monooctanoyl-l-Lysine

Yasuda

Kimura

Miyama

et al. 1989

Agricultural and Biological Chemistry

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Kinetics and mechanism of acyl transfer by penicillin acylases

Cited by 34 publications

References 38 publications

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic βLactam Antibiotics

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic βLactam Antibiotics

Penicillin acylase mutants with altered site-directed activity from Kluyvera citrophila

Polymyxin Acylase: An Enzyme Causing IntramolecularN²⇄N⁶Acyl Transfer inN-Monooctanoyl-l-Lysine

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Kinetics and mechanism of acyl transfer by penicillin acylases

Cited by 34 publications

References 38 publications

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic &#946;Lactam Antibiotics

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic &#946;Lactam Antibiotics

Penicillin acylase mutants with altered site-directed activity from Kluyvera citrophila

Polymyxin Acylase: An Enzyme Causing IntramolecularN2⇄N6Acyl Transfer inN-Monooctanoyl-l-Lysine

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Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic βLactam Antibiotics

Recent Advances in the Industrial Enzymatic Synthesis of Semi-Synthetic βLactam Antibiotics

Polymyxin Acylase: An Enzyme Causing IntramolecularN²⇄N⁶Acyl Transfer inN-Monooctanoyl-l-Lysine