Topic
Esterase
About: Esterase is a research topic. Over the lifetime, 7622 publications have been published within this topic receiving 168270 citations.
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TL;DR: The gene encoding an esterase from Pseudomonas fluorescens was subjected to random mutagenesis by error-prone PCR or by using the mutator strain Epicurian coli XL1-Red, resulting in the identification of several mutants showing up to almost two-fold enantioselectivity.
Abstract: The gene encoding an esterase from Pseudomonas fluorescens (PFE) was subjected to random mutagenesis by error-prone PCR or by using the mutator strain Epicurian coli XL1-Red. Enzyme libraries were then created in microtiter plates by expression of PFE-variants in E. coli. These were assayed for improved enantioselectivity in a Beckman robot system using optically pure (R)- or (S)-3-phenylbutyric acid resorufin esters, resulting in the identification of several mutants showing up to almost two-fold enantioselectivity (E(true) = 5.2 to 6.6) compared to wild-type PFE (E(true) = 3.5).
101 citations
TL;DR: These studies confirmed the quantitative changes in E4 protein previously reported and established that the increased esterase activity in P. humuli also arises from the production of more protein, or proteins, homologous to E4.
Abstract: An antiserum was prepared against carboxylesterase E4, the enzyme conferring resistance in Myzus persicae (Sulzer) to a wide range of insecticides, and the immunoglobulin G (IgG) fraction was purified from it by affinity chromotography. Interactions of the antiserum and IgG with aphid homogenates and the purified esterase proteins were studied by immune diffusion, immunoelectrophoresis and enzyme-linked immunosorbent assay (ELISA). In M. persicae, the interactions were specific for E4 and its closely-related mutant form, FE4, and except for Phorodon humuli (Schrank), there was no cross-reaction with homogenates of the nine other aphid species examined. These studies confirmed the quantitative changes in E4 protein previously reported and established that the increased esterase activity in P. humuli also arises from the production of more protein, or proteins, homologous to E4. Resistance of M. persicae could be characterized by immunoelectrophoresis even after preservation of the insects in 30% ethanol. Although ELISA could also be used to identify resistance, a simpler immunoplate assay was developed based on measuring the esterase activity of E4 retained when the enzyme bound to IgG. This assay discriminates well between the three resistant M. persicae variants common in the field in the UK, and its simplicity allows the study of large numbers of insects.
101 citations
TL;DR: It is demonstrated that tetraFA can be used to determine whether MeSA and its corresponding esterase(s) play a role in SAR signaling in other plant species, and the importance of SABP2 and MeSA for SAR development in tobacco is confirmed and similar roles are established.
101 citations
TL;DR: By analogy with the mechanism of trypsin action, trifluoromethylketones are probably potent inhibitors due to their resemblance to a tetrahedral transition state on the reaction coordinate to the acylated enzyme.
101 citations
TL;DR: The hypothesis that ester dependent lysosomal rupture is mediated by the specific interaction of the ester with a structural or functional lysOSomal protein is supported.
100 citations