Abstract:-The main goal of this research was the synthesis of enantiopure R(-)-3-aminoisobutyric acid from dihydrothymine with good yield, high stereospecificity and relative simplicity. Seventy two percent yield of the product was obtained in three steps.Step one consisted of dihydrothymine racemization.Step two was a dihydropyrimidinase reaction involving the Pseudomonas aeruginosa 10145 bacterial strain as the biocatalyst.Step three was performed with a diazotization reaction. The bacteria's enzymes determined the s… Show more
“…We first set up a one-pot cascade reaction with benzyl 3oxobutanoate, using similar conditions as those used for 3-OTfBE (see Figure 6), as it was one of the preferred β-keto esters for both hydrolase and transaminase reactions from those seven tested, and the possible end β-amino acid product of the reaction is an important intermediate for the production of biomedicals. [36] The products were quantified using HPLC. We found that the β-keto ester benzyl 3oxobutanoate (concentration, 14 mM) was converted to a high extent (> 95 % after 20 min and 99 % after 60 min; Figure S15).…”
Section: Methodsmentioning
confidence: 99%
“…[22,23] Simulation of the reaction by a local PELE exploration showed efficient 3-ATfBE and 3-OTfBE catalytic (hydrolytic) binding poses: 23 catalytic events for 3-ATfBE and 42 for 3-OTfBE when the serine-substrate distance threshold was set to 4 Å (Figure S12). Glyceryl tripropionate, an example of an ester commonly hydrolyzed by most esterases, [36] had 121 catalytic events with an associated experimental k cat of 388.1 min À 1 (Table S5). Thus, we expected that 3-OTfBE would be preferentially hydrolyzed.…”
Engineering dual-function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR 2 E 2 , with efficient native transaminase (k cat : 69.49 � 1.77 min À 1 ) and artificial esterase (k cat : 3908-0.41 min À 1 ) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR 2 E 2 (pH opt : 8.0-9.5; T opt : 60-65 °C) efficiently converts methyl 3-oxo-4-(2,4,5-trifluorophenyl)butanoate into 3-(R)-amino-4-(2,4,5-trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the β-keto ester into the β-keto acid at the hydrolytic site and subsequently into the β-amino acid (e.e. > 99 %) at the transaminase site. The catalytic power of the TR 2 E 2 PluriZyme was proven with a set of β-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.
“…We first set up a one-pot cascade reaction with benzyl 3oxobutanoate, using similar conditions as those used for 3-OTfBE (see Figure 6), as it was one of the preferred β-keto esters for both hydrolase and transaminase reactions from those seven tested, and the possible end β-amino acid product of the reaction is an important intermediate for the production of biomedicals. [36] The products were quantified using HPLC. We found that the β-keto ester benzyl 3oxobutanoate (concentration, 14 mM) was converted to a high extent (> 95 % after 20 min and 99 % after 60 min; Figure S15).…”
Section: Methodsmentioning
confidence: 99%
“…[22,23] Simulation of the reaction by a local PELE exploration showed efficient 3-ATfBE and 3-OTfBE catalytic (hydrolytic) binding poses: 23 catalytic events for 3-ATfBE and 42 for 3-OTfBE when the serine-substrate distance threshold was set to 4 Å (Figure S12). Glyceryl tripropionate, an example of an ester commonly hydrolyzed by most esterases, [36] had 121 catalytic events with an associated experimental k cat of 388.1 min À 1 (Table S5). Thus, we expected that 3-OTfBE would be preferentially hydrolyzed.…”
Engineering dual-function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR 2 E 2 , with efficient native transaminase (k cat : 69.49 � 1.77 min À 1 ) and artificial esterase (k cat : 3908-0.41 min À 1 ) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR 2 E 2 (pH opt : 8.0-9.5; T opt : 60-65 °C) efficiently converts methyl 3-oxo-4-(2,4,5-trifluorophenyl)butanoate into 3-(R)-amino-4-(2,4,5-trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the β-keto ester into the β-keto acid at the hydrolytic site and subsequently into the β-amino acid (e.e. > 99 %) at the transaminase site. The catalytic power of the TR 2 E 2 PluriZyme was proven with a set of β-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.
Engineering dual-function single polypeptide catalysts with two abiotic or biotic catalytic entities (or combinations of both) supporting cascade reactions is becoming an important area of enzyme engineering and catalysis. Herein we present the development of a PluriZyme, TR 2 E 2 , with efficient native transaminase (k cat : 69.49 � 1.77 min À 1 ) and artificial esterase (k cat : 3908-0.41 min À 1 ) activities integrated into a single scaffold, and evaluate its utility in a cascade reaction. TR 2 E 2 (pH opt : 8.0-9.5; T opt : 60-65 °C) efficiently converts methyl 3-oxo-4-(2,4,5-trifluorophenyl)butanoate into 3-(R)-amino-4-(2,4,5-trifluorophenyl)butanoic acid, a crucial intermediate for the synthesis of antidiabetic drugs. The reaction proceeds through the conversion of the β-keto ester into the β-keto acid at the hydrolytic site and subsequently into the β-amino acid (e.e. > 99 %) at the transaminase site. The catalytic power of the TR 2 E 2 PluriZyme was proven with a set of β-keto esters, demonstrating the potential of such designs to address bioinspired cascade reactions.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.