Vibrio mimicus Arylesterase Has Thioesterase and Chymotrypsin-like Activity

Chang, Rey-Chang; Chen, Jack Chien; Shaw, Jei‐Fu

doi:10.1006/bbrc.1995.2156

Cited by 13 publications

(4 citation statements)

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“…Hydrolysis of L-NBPNPE indicates that EstA has chymotrypsin-like activity. These results are similar to those observed for TesI from E. coli and EtpA from V. mimicus, which also exhibited chymotrypsin-like activity against L-NBPNPE (Chang et al 1995;Lee et al 1997).…”

Section: Discussionsupporting

confidence: 87%

Characterization of an arylesterase from Lactobacillus helveticus CNRZ32

2000

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An esterase gene (estA) was isolated from a previously constructed genomic library of Lactobacillus helveticus CNRZ32. The estA gene consisted of a 558 bp open reading frame encoding a putative peptide of 21·3 kDa. Protein sequence homology searches using BLAST revealed that EstA had low amino acid sequence identity with the serine‐dependent arylesterases TesI (24%) and EtpA (26%) from Escherichia coli and Vibrio mimicus, respectively. A recombinant EstA fusion protein containing a C‐terminal six‐histidine tag was constructed and purified to electrophoretic homogeneity. Characterization of EstA revealed that it was a serine‐dependent enzyme having a monomeric Mr of 22·6–25·1 kDa. Optimum temperature, NaCl concentration and pH for EstA activity were determined to be 35–40 °C, 3·5% NaCl and 7·5–8·0, respectively. EstA had significant activity under conditions simulating those of ripening cheese (10 °C, 4% NaCl, pH 5·1). EstA hydrolysed a variety of ester compounds and preferred those with substituted phenyl alcohol and short‐chain fatty acid groups. Site‐directed mutagenesis suggested that the S10 and H164 residues were essential for EstA activity.

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Section: Discussionsupporting

confidence: 87%

Characterization of an arylesterase from Lactobacillus helveticus CNRZ32

2000

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“…Furthermore, due to the flexibility of its catalytic domain [9], the SGNH superfamily has multifunctional properties and broad specificities for substrates including carbohydrate esterase [10][11][12], lipase [13], protease [14], thioesterase [15,16], arylesterase [15], lysophospholipase [17], and acyltransferase [13,14]. The SGNH hydrolases from plants are involved in the regulation of development and morphogenesis [18,19], defense [20], as well as tolerance to environmental stresses [21].…”

Section: Introductionmentioning

confidence: 99%

Structure guided protein engineering increases enzymatic activities of the SGNH family esterases

Huo

et al. 2020

Preprint

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Background Esterases and lipases hydrolyze short-chain esters and long-chain triglycerides, respectively, and therefore play essential roles in the synthesis and decomposition of ester bonds in the pharmaceutical and food industries. Many SGNH family esterases share high similarity in sequences. However, they have distinct enzymatic activities toward the same substrates. Due to a lack of structural information, the detailed catalytic mechanisms of these esterases remain barely investigated. Results In this study, we identified two SGNH family esterases, CrmE10 and AlinE4, from marine bacteria with significantly different preferences for pH, temperature, metal ion, and organic solvent tolerance despite high sequence similarity. The crystal structures of these two esterases, including wild type and mutants, were determined to high resolutions ranging from 1.18 Å to 2.24 Å. Both CrmE10 and AlinE4 were composed of five β-strands and nine α-helices, which formed one compact N-terminal α/β globular domain and one extended C-terminal domain. The aspartic residues (D178 in CrmE10/ D162 in AlinE4) stabilized the conformations of the catalytic triad (Ser-Asp-His) in both esterases, and the metal ion Cd2+ might reduce enzymatic activity by blocking proton transfer or substrate binding. CrmE10 and AlinE4 showed distinctly different electrostatic surface potentials, despite the similar atomic architectures and a similar swap catalytic mechanism. When five negative charged residues (Asp or Glu) were mutated to residue Lys, CrmE10 obtained elevated alkaline-adaptability and significantly increased the enzymatic activity from 0% to 20% at pH 10.5. Also, CrmE10 mutants exhibited dramatic change for enzymatic properties when compared with the wide-type enzyme. Conclusions These findings offer a perspective for understanding the catalytic mechanism of different esterases and might facilitate the industrial biocatalytic applications.

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“…The SGNH superfamily has conserved homologs ranging from prokaryotes to eukaryotes. Furthermore, due to the exibility of its catalytic domain [9], the SGNH superfamily has multifunctional properties and broad speci cities for substrates including carbohydrate esterase [10][11][12], lipase [13], protease [14], thioesterase [15,16], arylesterase [15], lysophospholipase [17], and acyltransferase [13,14]. The SGNH hydrolases from plants are involved in the regulation of development and morphogenesis [18,19], defense [20], as well as tolerance to environmental stresses [21].…”

Section: Introductionmentioning

confidence: 99%

Structure guided protein engineering increases enzymatic activities of the SGNH family esterases

Huo

et al. 2020

Preprint

View full text Add to dashboard Cite

Background Esterases and lipases hydrolyze short-chain esters and long-chain triglycerides, respectively, and therefore play essential roles in the synthesis and decomposition of ester bonds in the pharmaceutical and food industries. Many SGNH family esterases share high similarity in sequences. However, they have distinct enzymatic activities toward the same substrates. Due to a lack of structural information, the detailed catalytic mechanisms of these esterases remain barely investigated. Results In this study, we identified two SGNH family esterases, CrmE10 and AlinE4, from marine bacteria with significantly different preferences for pH, temperature, metal ion, and organic solvent tolerance despite high sequence similarity. The crystal structures of these two esterases, including wild type and mutants, were determined to high resolutions ranging from 1.18 Å to 2.24 Å. Both CrmE10 and AlinE4 were composed of five β-strands and nine α-helices, which formed one compact N-terminal α/β globular domain and one extended C-terminal domain. The aspartic residues (D178 in CrmE10/ D162 in AlinE4) destabilized the conformations of the catalytic triad (Ser-Asp-His) in both esterases, and the metal ion Cd2+ might reduce enzymatic activity by blocking proton transfer or substrate binding. CrmE10 and AlinE4 showed distinctly different electrostatic surface potentials, despite the similar atomic architectures and a similar swap catalytic mechanism. When five negative charged residues (Asp or Glu) were mutated to residue Lys, CrmE10 obtained elevated alkaline-adaptability and significantly increased the enzymatic activity from 0% to 20% at pH 10.5. Also, CrmE10 mutants exhibited dramatic changes for enzymatic properties when compared with the wide-type enzyme. Conclusions These findings offer a perspective for understanding the catalytic mechanism of different esterases and might facilitate the industrial biocatalytic applications.

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Vibrio mimicus Arylesterase Has Thioesterase and Chymotrypsin-like Activity

Cited by 13 publications

References 0 publications

Characterization of an arylesterase from Lactobacillus helveticus CNRZ32

Characterization of an arylesterase from Lactobacillus helveticus CNRZ32

Structure guided protein engineering increases enzymatic activities of the SGNH family esterases

Structure guided protein engineering increases enzymatic activities of the SGNH family esterases

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