What makes a good new therapeutic l-asparaginase?

Beckett, Angela H.; Gervais, David

doi:10.1007/s11274-019-2731-9

Cited by 84 publications

(57 citation statements)

References 111 publications

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“…Most of l -asparaginases also catalyze the hydrolysis of l -Gln to l -Glu, and those with glutaminase activity comparable or higher than asparaginase activity are often referred to as glutaminases-asparaginases (EC 3.5.1.38). Two structurally related l -asparaginases are present in the model bacterium Escherichia coli , with their names abbreviated EcAI (for cytoplasmic enzyme) and EcAII (for periplasmic enzyme).Asparaginases from two bacterial sources (EcAII from E. coli and ErA from Erwinia chrysanthemi ( Dickeya dadantii )) came to a focus in 1960s–1970s, after being identified as successful agents in treatment of various leukemias and lymphomas, prominently the juvenile non-Hodgkin, lymphoblastic leukemia (ALL)—the most prevalent pediatric cancer (reviewed in 2 – 4 ). Effectiveness of l -asparaginases is based on a principle of amino acid restriction.…”

Section: Introductionmentioning

confidence: 99%

“…Due to their efficacy and to economic factors 5 , l -asparaginases remain the first-line drugs in the treatment of ALL, despite recent developments of immunotherapies 8 , 9 . Several modified preparations of these enzymes with reduced immunogenicity and increased stability, have been developed since addition of l -asparaginases to the therapeutic repertoire 2 . Nevertheless, despite all advances and a high rate of therapeutic success in ALL, several deficiencies of asparaginase therapy have been reported.…”

Section: Introductionmentioning

confidence: 99%

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Generalized enzymatic mechanism of catalysis by tetrameric l-asparaginases from mesophilic bacteria

Strzelczyk

Zhang

Dyba

et al. 2020

Sci Rep

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The mechanism of catalysis by the l-glutaminase-asparaginase from Pseudomonas 7A (PGA) was investigated using structural, mass spectrometry, and kinetic data. We had previously proposed mechanism of hydrolysis of l-Asn by the type II l-asparaginase from E. coli (EcAII), but that work was limited to just one enzyme. Based on results presented in this report, we postulate that all homotetrameric l-asparaginases from mesophilic bacteria utilize a common ping-pong mechanism of catalysis consisting of two subsequent nucleophilic substitutions. Several new structures of non-covalent complexes of PGA with different substrates, as well as structures of covalent acyl-enzyme intermediates of PGA with canonical substrates (l-Asp and l-Glu) and an opportunistic ligand, a citrate anion, were determined. The results of kinetic experiments monitored by high-resolution LC/MS, when combined with new structural data, clearly show that the reaction catalyzed by l-glutaminase-asparaginases proceeds through formation of a covalent intermediate, as observed previously for EcAII. Additionally, by showing that the same mechanism applies to l-Asn and l-Gln, we postulate that it is common for all these structurally related enzymes.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Generalized enzymatic mechanism of catalysis by tetrameric l-asparaginases from mesophilic bacteria

Strzelczyk

Zhang

Dyba

et al. 2020

Sci Rep

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show abstract

“…Additionally, the representatives of the 'Candidatus Rokubacteria' are ubiquitous in a diverse range of terrestrial ecosystems and subsurface habitats with no reported marine representative [59]. CAspII and CAspIII also revealed catalytic e ciencies of approximately 10-fold higher than CAspI suggesting that they could metabolize asparagine more e ciently [61]. Effect of pH and NaCl concentration on enzyme activity.…”

Section: Resultsmentioning

confidence: 99%

Metagenomic discovery and functional validation of L-asparaginases with anti-leukemic effect from the Caspian Sea

Sobat

Asad

Kabiri

et al. 2020

Preprint

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Background L-asparaginase has been used for the treatment of acute lymphoblastic leukemia (ALL) for more than 30 years. However, efforts continued to find new enzymes with more desirable properties due to the immunogenicity, short half-life, rapid clearance and L-glutaminase side activity of the existing commercial enzymes. Screening for novel L-asparaginases in prokaryotes as a promising resource has been mainly hampered by the cultivation/expression bottleneck. Results By screening 27000 publicly available prokaryotic genomes, we recovered ca. 6300 type I and ca. 5200 type II putative L-asparaginase in 36 and 42 bacterial phyla respectively highlighting the vast potential of prokaryotes for L-asparaginase activity. Caspian water with similar salt composition to the human serum was targeted for in-silico screening of L-asparaginase. We screened ca. three million predicted Open Reading Frames of the assembled Caspian Sea metagenomes. In-silico screening resulted in 87 putative L-asparaginase genes from the Caspian Sea datasets. The L-asparagine hydrolysis was experimentally confirmed by cloning three selected genes (1092, 1218 and 1011 bp) in E. coli. Catalytic parameters of the purified enzymes including Km, Vmax and catalytic efficiency were determined to be among the most desirable reported values of microbial L-asparaginases. Two of the recombinant enzymes represented remarkable anti-proliferative activity (IC50 less than 1 IU/ml) against leukemia cell line Jurkat while no cytotoxic effect on human erythrocytes or human umbilical vein endothelial cells was detected. Conclusions Similar salinity and ionic concentration of the Caspian water samples to the human serum highlights the secretory L-asparaginases recovered from these metagenomes as potential treatment agents.

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“…Additionally, the representatives of the ''Candidatus Rokubacteria'' are ubiquitous in a diverse range of terrestrial ecosystems and subsurface habitats with no reported marine representatives (Becraft et al, 2017). CAspII and CAspIII also revealed catalytic efficiencies of approximately 10-fold higher than CAspI suggesting that they could metabolize asparagine more efficiently (Beckett and Gervais, 2019).…”

Section: Kinetic Parameters Of Recombinant L-asparaginases Were Among the Most Desirable Reported Values Of Microbial L-asparaginasesmentioning

confidence: 99%

Metagenomic discovery and functional validation of L-asparaginases with anti-leukemic effect from the Caspian Sea

et al. 2021

View full text Add to dashboard Cite

By screening 27,000 publicly available prokaryotic genomes, we recovered ca. 6300 type I and ca. 5200 type II putative L-asparaginase highlighting the vast potential of prokaryotes. Caspian water with similar salt composition to the human serum was targeted for in silico L-asparaginase screening. We screened ca. three million predicted genes of its assembled metagenomes that resulted in annotation of 87 putative L-asparaginase genes. The L-asparagine hydrolysis was experimentally confirmed by synthesizing and cloning three selected genes in E. coli. Catalytic parameters of the purified enzymes were determined to be among the most desirable reported values. Two recombinant enzymes represented remarkable anti-proliferative activity (IC50 <1IU/ml) against leukemia cell line Jurkat while no cytotoxic effect on human erythrocytes or human umbilical vein endothelial cells was detected. Similar salinity and ionic concentration of the Caspian water to the human serum highlights the potential of secretory L-asparaginases recovered from these metagenomes as potential treatment agents.

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What makes a good new therapeutic l-asparaginase?

Cited by 84 publications

References 111 publications

Generalized enzymatic mechanism of catalysis by tetrameric l-asparaginases from mesophilic bacteria

Generalized enzymatic mechanism of catalysis by tetrameric l-asparaginases from mesophilic bacteria

Metagenomic discovery and functional validation of L-asparaginases with anti-leukemic effect from the Caspian Sea

Metagenomic discovery and functional validation of L-asparaginases with anti-leukemic effect from the Caspian Sea

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