The quantitation of rat serum esterases by densitometry of acrylamide gels stained for enzyme activity

Rosenberg, Martin; Roegner, Vivien; Becker, Frederick F.

doi:10.1016/0003-2697(75)90738-1

Cited by 38 publications

(16 citation statements)

References 16 publications

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“…(1992a), was plated in 0.7% (w/v) agarose overlays containing 10 mM IPTG to induce gene expression using E. coli XL1-Blue cells as the host strain. Esterase activity was detected by a method adapted from that described by Rosenberg et al (1975). The plaques were transferred to Whatman no.…”

Section: Methodsmentioning

confidence: 99%

“…However, a number of acetylxylan esterases have been reported as having activity against a-naphthyl acetate (a-NA) and other artificial substrates (McDermid et al, 1990;Sundberg & Poutanen, 1991). A convenient method for detecting such esterase activity using general esterase substrates such as a-NA was described by Rosenberg et al (1975). Although not all esterases that hydrolyse such substrates are acetylxylan esterases, it is likely that under conditions of high levels of synthesis of fibre-degrading enzymes, a significant proportion of esterases able to hydrolyse a-NA, or similar compounds, may be acetylxylan esterases.…”

Section: Introductionmentioning

confidence: 99%

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Three Neocallimastix patriciarum esterases associated with the degradation of complex polysaccharides are members of a new family of hydrolases

Dalrymple

Cybinski

et al. 1997

Microbiology

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Acetylesterase and cinnamoyl ester hydrolase activities were demonstrated in culture supernatant of the anaerobic ruminal fungus Neocallimastix patriciarum. A cDNA expression library from N. patriciarum was screened for esterases using p-naphthyl acetate and a model cinnamoyl ester compound. cDNA clones representing four different esterase genes (bnaA-D) were isolated. None of the enzymes had cinnamoyl ester hydrolase activity, but two of the enzymes (BnaA and BnaC) had acetylxylan esterase activity. bnaA, bnaB and bnaC encode proteins with several distinct domains. Carboxy-terminal repeats in BnaA and BnaC are homologous to protein-docking domains in other enzymes from Neocallimastix species and another anaerobic fungus, a Piromyces sp. The catalytic domains of BnaB and BnaC are members of a recently described family of Ser/His active site hydrolases [Upton, C. & Buckley, J. T. (1995). Trends Biochem Sci 20, 178-1791. BnaB exhibits 40% amino acid identity to a domain of unknown function in the CelE cellulase from Clostridium thermocellum and BnaC exhibits 52% amino acid identity to a domain of unknown function in the XynB xylanase from Ruminococcus flavefaciens. BnaA, whilst exhibiting less than 10 O/ O overall amino acid identity to BnaB or BnaC, or to any other known protein, appears to be a member of the same family of hydrolases, having the three universally conserved amino acid sequence motifs. Several other previously described esterases are also shown to be members of this family, including a rhamnogalacturonan acetylesterase from Aspergillus aculeatus. However, none of the other previously described enzymes with acetylxylan esterase activity are members of this family of hydrolases.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Three Neocallimastix patriciarum esterases associated with the degradation of complex polysaccharides are members of a new family of hydrolases

Dalrymple

Cybinski

et al. 1997

Microbiology

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“…Brominating activity was detected by the conversion of the substrate phenol red to bromophenol blue (Loo et al, 1964). Esterase activity was detected in situ using the procedure described by Rosenberg et al (1975), with phenyl acetate or 2-naphthyl acetate as substrates and Diazo Blue B as coupling agent.…”

Section: Introductionmentioning

confidence: 99%

A bacterial esterase is homologous with nonhaem haloperoxidases and displays brominating activity

Pelletier

Altenbuchner

1995

Microbiology

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Screening GenBank indicated that an esterase from Pseudomonas fluorescens had high sequence similarity with bacterial non-haem haloperoxides. However, this homology was limited to two distinct domains of the published esterase sequence. As errors in the published sequence were suspected, the esterase gene was sequenced again. The revised sequence displayed between 40 and 50% identical amino acids with the haloperoxidases, but distributed along the whole sequence. In addition to the structural homologies with haloperoxidases, the esterase also displayed functional homology. The recombinant esterase, purified from Escherichia coli cells, was capable of both ester hydrolysis and halogenation, as detected in situ by the formation of bromophenol blue or spectrophotometrically by the bromination of monochlorodimedon. The esterase is thus a bifunctional enzyme. The sequence analysis and the biochemical investigations show that the esterase belongs to the haloperoxidase family. It also possessed, however, a typical feature of serine-hydrolases, namely the consensus motif Gly-X-Ser-X-Gly around the active serine of the catalytic triad. By alignment of the esterase with different serine-hydrolase sequences, it was possible to identify the other two residues of the triad. The triad comprised the residues Ser95, Asp223 and His252. Interestingly, a structurally equivalent catalytic triad was also identified in the sequences of all bacterial non-haem haloperoxidases, in highly conserved domains. The presence of a catalytic triad in haloperoxidases is expected to be important in the mechanism of halogenation.

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“…In some cases, esterases exhibiting xylan deacetylating activity as a side reaction only were assigned as acetyl xylan esterases, such as the CE2 esterases (see below). General esterase activity assays, using α-naphtyl acetate [28] and 4-nitrophenyl acetate as substrates, have frequently been used to determine AcXE activity [19], but cannot distinguish AcEs from AcXEs. AcEs differ from AcXEs in that AcEs are generally not capable of deacetylating polymeric or oligomeric xylan.…”

Section: Acetyl Xylan Esterases (Acxes)mentioning

confidence: 99%

Phylogeny, classification and metagenomic bioprospecting of microbial acetyl xylan esterases

Adesioye

Makhalanyane

Biely

et al. 2016

Enzyme and Microbial Technology

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Highlights• AcXEs occur in several CAZy families, and show promiscuous substrate specificities.• A strong AcXE sequence to specificity link would be a valuable functional predictor.• There is a clear need for novel AcXE enzymes with detailed substrate specificity data.• Functional metagenomics would be the most effective means for identifying new AcXEs.• A lack of suitable substrates limits high throughput metagenomic biomining of AcXEs.

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The quantitation of rat serum esterases by densitometry of acrylamide gels stained for enzyme activity

Cited by 38 publications

References 16 publications

Three Neocallimastix patriciarum esterases associated with the degradation of complex polysaccharides are members of a new family of hydrolases

Three Neocallimastix patriciarum esterases associated with the degradation of complex polysaccharides are members of a new family of hydrolases

A bacterial esterase is homologous with nonhaem haloperoxidases and displays brominating activity

Phylogeny, classification and metagenomic bioprospecting of microbial acetyl xylan esterases

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