A Product Analog Bound Form of 3-Oxoadipate-enol-Lactonase (PcaD) Reveals a Multifunctional Role for the Divergent Cap Domain

Bains, Jasleen Kaur; Kaufman, Laura; Farnell, Benjamin; Boulanger, Martin J.

doi:10.1016/j.jmb.2011.01.007

Cited by 37 publications

(53 citation statements)

References 52 publications

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“…OELH catalyzes the important conversion of cyclic 3-oxoadipate enollactone to linear 3-oxoadipate, which is finally transformed to citric acid cycle intermediates succinyl-CoA and acetyl-CoA. Structural analysis of the OELH from the proteobacterium Burkholderia xenovorans found in dimeric form suggested the catalytic mechanism (Bains, Kaufman, Farnell, & Boulanger, 2011).…”

Section: Enzymes Of the Ortho-ring Cleavage Pathwaysmentioning

confidence: 99%

Catabolism of Phenol and Its Derivatives in Bacteria

Nešvera

Rucká

Pátek

2015

Advances in Applied Microbiology

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Section: Enzymes Of the Ortho-ring Cleavage Pathwaysmentioning

confidence: 99%

Catabolism of Phenol and Its Derivatives in Bacteria

Nešvera

Rucká

Pátek

2015

Advances in Applied Microbiology

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“…While it is unlikely that the exact function of distant homologs is the same, the affinity towards large hydrophobic epoxides and conservation of catalytic mechanism (as evidenced by active site superposition - Figure 7) are likely. Presence of several conserved arginines within the cap domain raises possibility of their involvement in substrate binding or orientation (coupled with conformational change), analogous to the mechanisms observed previously in dienelactone hydrolase [20] and 3-oxoadipate enol lactonase [16]. Elucidation of the full substrate orientation/catalysis scenario (including involvement of the glutamate and aspartate residues and their spatial conformations during the process) is planned through application of molecular dynamics experiments for modelling of the ligand binding process.…”

Section: Discussionmentioning

confidence: 73%

“…While homology-based models are likely insufficient for elucidation of full sequence of events during substrate binding and catalysis (both the variable cap domain e.g. [16,17] and surrounding loops [18] are involved in controlling and fine-tuning substrate access), we were nevertheless able to ascertain the key functional residues involved.…”

Section: Resultsmentioning

confidence: 99%

Zearalenone lactonohydrolase activity in Hypocreales and its evolutionary relationships within the epoxide hydrolase subset of a/b-hydrolases

et al. 2014

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BackgroundZearalenone is a mycotoxin produced by several species of Fusarium genus, most notably Fusarium graminearum and Fusarium culmorum. This resorcylic acid lactone is one of the most important toxins causing serious animal and human diseases. For over two decades it has been known that the mycoparasitic fungus Clonostachys rosea (synonym: Gliocladium roseum, teleomorph: Bionectria ochroleuca) can detoxify zearalenone, however no such attributes have been described within the Trichoderma genus.ResultsWe screened for the presence of zearalenone lactonohydrolase homologs in isolates of Clonostachys and Trichoderma genera. We report first finding of expressed zearalenone lactonohydrolase in Trichoderma aggressivum. For three isolates (T. aggressivum, C. rosea and Clonostachys catenulatum isolates), we were able to reconstruct full coding sequence and verify the biotransformation ability potential. Additionally, we assessed progression of the detoxification process (in terms of transcript accumulation and mycotoxin decomposition in vitro).In silico, search for origins of zearalenone lactonohydrolase activity in model fungal and bacterial genomes has shown that zearalenone lactonohydrolase homologs form a monophyletic fungal clade among the a/b hydrolase superfamily representatives. We corroborated the finding of functional enzyme homologs by investigating the functional sites (active site pocket with postulated, noncanonical Ser-Glu-His catalytic triad) conserved in both multiple sequence alignment and in homology-based structural models.ConclusionsOur research shows the first finding of a functional zearalenone lactonohydrolase in mycoparasitic Trichoderma aggressivum (an activity earlier characterised in the Clonostachys rosea strains). The supporting evidence for presence and activity of functional enzyme homologs is based on the chemical analyses, gene expression patterns, homology models showing conservation of key structural features and marked reduction of zearalenone content in cultured samples (containing both medium and mycelium). Our findings also show divergent strategies of zearalenone biotransformation ability (rapid induced expression and detoxification vs. gradual detoxification) present in several members of Hypocreales order (Trichoderma and Clonostachys genera). The potential for lactonhydrolase activity directed towards zearalenone and/or similar compounds is likely ancient, with homologs present in several divergent filamentous fungi among both Sordariomycetes (Bionectria sp., Trichoderma sp., Apiospora montagnei) and Leotiomycetes (Marssonina brunnea f. sp. ‘multigermtubi’).

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“…Addition, a BLAST search of HydS14 (BAM08272) revealed 54% identity to 3-oxoadipate enol-lactone hydrolases from S. erythraea NRRL 2338 (YP_001103223). Interestingly, 3-oxoadipate enol-lactone hydrolases (EC 3.1.1.24), which are key enzymes in the β-ketoadipate pathway for the dissimilation of aromatic compounds, are included in this same group cluster [17]. α/β-Hydrolase-fold enzymes include various enzyme groups with diverse catalytic functions [18], including diene lactone hydrolases and lipases.…”

Section: Resultsmentioning

confidence: 99%

Cloning, Expression and Characterization of a Thermostable Esterase HydS14 from Actinomadura sp. Strain S14 in Pichia pastoris

Sriyapai

Kawai

Siripoke

et al. 2015

IJMS

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A thermostable esterase gene (hydS14) was cloned from an Actinomadura sp. S14 gene library. The gene is 777 bp in length and encodes a polypeptide of 258 amino acid residues with no signal peptide, no N-glycosylation site and a predicted molecular mass of 26,604 Da. The encoded protein contains the pentapeptide motif (GYSLG) and catalytic triad (Ser88-Asp208-His235) of the esterase/lipase superfamily. The HydS14 sequence shows 46%–64% identity to 23 sequences from actinomycetes (23 α/β-hydrolases), has three conserved regions, and contains the novel motif (GY(F)SLG), which distinguishes it from other clusters in the α/β-hydrolase structural superfamily. A plasmid containing the coding region (pPICZαA-hydS14) was used to express HydS14 in Pichia pastoris under the control of the AOXI promoter. The recombinant HydS14 collected from the supernatant had a molecular mass of ~30 kDa, which agrees with its predicted molecular mass without N-glycosylation. HydS14 had an optimum temperature of approximately 70 °C and an optimum pH of 8.0. HydS14 was stable at 50 and 60 °C for 120 min, with residual activities of above 80% and above 90%, respectively, as well as 50% activity at pH 6.0–8.0 and pH 9.0, respectively. The enzyme showed higher activity with p-nitrophenyl-C2 and C4. The Km and Vmax values for p-nitrophenyl-C4 were 0.21 ± 0.02 mM and 37.07 ± 1.04 μmol/min/mg, respectively. The enzyme was active toward short-chain p-nitrophenyl ester (C2–C6), displaying optimal activity with p-nitrophenyl-C4 (Kcat/Km = 11.74 mM−1·S−1). In summary, HydS14 is a thermostable esterase from Actinomadura sp. S14 that has been cloned and expressed for the first time in Pichia pastoris.

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A Product Analog Bound Form of 3-Oxoadipate-enol-Lactonase (PcaD) Reveals a Multifunctional Role for the Divergent Cap Domain

Cited by 37 publications

References 52 publications

Catabolism of Phenol and Its Derivatives in Bacteria

Catabolism of Phenol and Its Derivatives in Bacteria

Zearalenone lactonohydrolase activity in Hypocreales and its evolutionary relationships within the epoxide hydrolase subset of a/b-hydrolases

Cloning, Expression and Characterization of a Thermostable Esterase HydS14 from Actinomadura sp. Strain S14 in Pichia pastoris

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