Synthesis and spectral characterization of 2,2-diphenylethyl glucosinolate and HPLC-based reaction progress curve data for the enzymatic hydrolysis of glucosinolates by Sinapis alba myrosinase

Abstract

The data presented in this article are related to the research article, “HPLC-based enzyme kinetics assay for glucosinolate hydrolysis facilitate analysis of systems with both multiple reaction products and thermal enzyme denaturation” (C.K. Klingaman, M.J. Wagner, J.R. Brown, J.B. Klecker, E.H. Pauley, C.J. Noldner, J.R. Mays,) [1]. This data article describes (1) the synthesis and spectral characterization data of a non-natural glucosinolate analogue, 2,2-diphenylethyl glucosinolate, (2) HPLC standardization data for glucosinolate, isothiocyanate, nitrile, and amine analytes, (3) reaction progress curve data for enzymatic hydrolysis reactions with variable substrate concentration, enzyme concentration, buffer pH, and temperature, and (4) normalized initial velocities of hydrolysis/formation for analytes. These data provide a comprehensive description of the enzyme-catalyzed hydrolysis of 2,2-diphenylethyl glucosinolate (5) and glucotropaeolin (6) under widely varied conditions.

HPLC-based kinetics assay facilitates analysis of systems with multiple reaction products and thermal enzyme denaturation

“…Representative reaction progress curves at 227 nm are depicted in Fig. 4; progress curves for all other wavelengths are available [34]. Similar to previous accounts [19], detectable [Gluc] t and [ITC] t did not change over time in the absence of enzyme.…”

“…Initial rates ( V 0 ) of glucosinolate hydrolysis (Δ[Gluc] Δ t −1 ) and ITC formation (Δ[ITC] Δ t −1 ) were obtained from their slopes at t = 0 min (see Tables 2 and 3 in Ref. [34]). Representative reaction progress curves monitored at 227 nm for the conversion of 5 to 8 are provided (Fig.…”

“…Representative reaction progress curves monitored at 227 nm for the conversion of 5 to 8 are provided (Fig. 2); progress curves for all other concentration/wavelength combinations are available [34]. For this substrate, [ITC] t data (Fig.…”

“…3; analogous plots for other wavelengths are available [34]. Each Michaelis–Menten curve was fit with a high correlation ( r 2 > 0.9903), and converged K m and V max values were consistent across independently monitored wavelengths, as evidenced by results from pooled wavelength data ( r 2 = 0.9911, Table 1) [19].…”

“…The V 0 scaled linearly with [Myr] ( r 2 > 0.9770) when monitoring both [Gluc] t and [ITC] t (Fig. 5); specific activity plots for all other wavelengths are available [34]. Specific activities of 5 versus the standard ([ 7 ] 0 = 250 µM, 0.94 min −1 ) ranged from 19–20%.…”