Presentation Title

An Amino Acid Switch Increases the Promiscuity of a Carbon-Carbon Bond Forming Biocatalyst

Faculty Mentor

Sherif I Elshahawi

Start Date

23-11-2019 8:45 AM

End Date

23-11-2019 9:30 AM

Location

206

Session

poster 2

Type of Presentation

Poster

Subject Area

interdisciplinary

Abstract

Enzymes catalyze chemical reactions under water-based physiological conditions. They offer a green alternative to synthetic reactions, which require toxic petroleum-derived solvents and extreme conditions of temperature and/or pH. Furthermore, engineering of enzymes usually results in either improving the native function of the enzyme or even allow them to perform new-to-nature chemical reactions. Modification of compounds is a significant approach to the development of new chemical entities and biological activities. Prenyltransferases (PT) are late-stage tailoring enzymes that exist in living systems and are encoded in natural product biosynthetic gene clusters. PTs catalyze the attachment of a five-carbon unit called prenyl, to aromatic acceptors using pyrophosphate donors. This prenyl modification leads to changes in structural and biological activities in small molecules. Our lab has studied the active site of the crystal structure of a tryptophan specific PT that is capable of forming carbon-carbon bonds. A structural-guided enzyme engineering of the wild-type enzyme has led to the improvement in its activity. Using HPLC, nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectrometry (HR-MS), we show that the engineered enzyme has increased its substrate flexibility to accommodate tyrosine, an activity that is lacking in the wild type. The long-term goal of this study is to expand the engineered promiscuity of PT enzymes for drug diversification.

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Nov 23rd, 8:45 AM Nov 23rd, 9:30 AM

An Amino Acid Switch Increases the Promiscuity of a Carbon-Carbon Bond Forming Biocatalyst

206

Enzymes catalyze chemical reactions under water-based physiological conditions. They offer a green alternative to synthetic reactions, which require toxic petroleum-derived solvents and extreme conditions of temperature and/or pH. Furthermore, engineering of enzymes usually results in either improving the native function of the enzyme or even allow them to perform new-to-nature chemical reactions. Modification of compounds is a significant approach to the development of new chemical entities and biological activities. Prenyltransferases (PT) are late-stage tailoring enzymes that exist in living systems and are encoded in natural product biosynthetic gene clusters. PTs catalyze the attachment of a five-carbon unit called prenyl, to aromatic acceptors using pyrophosphate donors. This prenyl modification leads to changes in structural and biological activities in small molecules. Our lab has studied the active site of the crystal structure of a tryptophan specific PT that is capable of forming carbon-carbon bonds. A structural-guided enzyme engineering of the wild-type enzyme has led to the improvement in its activity. Using HPLC, nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectrometry (HR-MS), we show that the engineered enzyme has increased its substrate flexibility to accommodate tyrosine, an activity that is lacking in the wild type. The long-term goal of this study is to expand the engineered promiscuity of PT enzymes for drug diversification.