Direct Spectrophotometric Assay for Benzaldehyde Lyase Activity

Abstract

Benzaldehyde lyase from Pseudomonas fluorescens Biovar I. (BAL, EC 4.1.2.38) is a versatile catalyst for the organic synthesis of chiral α-hydroxy ketones. To allow fast assessment of enzyme activity, a direct spectrophotometric assay is desirable. Here, a new robust and easy-to-handle assay based on UV absorption is presented. The assay developed is based on the ligation of the α-hydroxy ketone (R)-2,2′-furoin from 2-furaldehyde. A robust assay with direct monitoring of the product is facilitated with a convenient concentration working range minimising experimental associated with low concentrations.

Modeling‐Assisted Design of Thermostable Benzaldehyde Lyases from Rhodococcus erythropolis for Continuous Production of α‐Hydroxy Ketones

“… [24] Furthermore, one potential BAL each was selected from Sphaerobacter thermophilus ( St BAL) and Thermobacillus composti ( Tc BAL). The enzymes were obtained by heterologous expression in Escherichia coli (Figure S1A, sequence data see Table S2) and the raw cell lysates were initially screened for aromatic ligase activity, using a literature‐reported photometric assay [25] based on the conversion of 2‐furaldehyde ( 1 a , Scheme 2 ) into 2,2’‐furoin ( 2 a ) (see also Figure S1B).…”

Cover Feature: Modeling‐Assisted Design of Thermostable Benzaldehyde Lyases from Rhodococcus erythropolis for Continuous Production of α‐Hydroxy Ketones (ChemBioChem 7/2022)

“…[24] Furthermore, one potential BAL each was selected from Sphaerobacter thermophilus (StBAL) and Thermobacillus composti (TcBAL). The enzymes were obtained by heterologous expression in Escherichia coli (Figure S1A, sequence data see Table S2) and the raw cell lysates were initially screened for aromatic ligase activity, using a literature-reported photometric assay [25] based on the conversion of 2-furaldehyde (1 a, Scheme 2) into 2,2'-furoin (2 a) (see also Figure S1B).…”