Design of a 3-steps D-DIBOA purification method without organic solvent wastes.
•Novel affinity chromatography for the purification of benzohydroxamic acids.
•Prototype for the automation of D-DIBOA purification from whole-cell biocatalysis.
•Both biological and synthetic D-DIBOA have the same herbicide activity.
AbstractHerbicides play a vital role in agriculture, contributing to increased crop productivity by minimizing weed growth, but their low degradability presents a threat to the environment and human health. Allelochemicals, such as DIBOA (2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4 H)-one), are secondary metabolites released by certain plants that affect the survival or growth of other organisms. Although these metabolites have an attractive potential for use as herbicides, their low natural production is a critical hurdle. Previously, the synthesis of the biologically active analog D-DIBOA (4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one) was achieved, using an engineered E. coli strain as a whole-cell biocatalyst, capable of transforming a precursor compound into D‐DIBOA and exporting it into the culture medium, although it cannot be directly applied to crops. Here a chromatographic method to purify D-DIBOA from this cell culture medium without producing organic solvent wastes is described. The purification of D-DIBOA from a filtered culture medium to the pure compound could also be automated. Biological tests with the purified compound on weed models showed that it has virtually the same activity than the chemically synthesized D-DIBOA.
AbbreviationsDIBOA2,4-dihydroxy-(2H)-1,4-benzoxazin-3-(4H)one
D-DIBOA4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one, or 2-deoxy-DIBOA
wcb-D-DIBOAD-DIBOA obtained by whole-cell biocatalysis
cs-D-DIBOAD-DIBOA obtained by chemical synthesis
PCethyl-2-(2´-nitrophenoxy)acetate (D-DIBOA precursor)
IMACimmobilized-metal affinity chromatography
IEXion-exchange chromatography
Mono Qanion exchange column
Mono Scation exchange column
RPLCReversed Phase Liquid Chromatography
KeywordsD-DIBOA
Green chemistry
Whole-cell biocatalyst
Downstream processes
Chromatography
Ecological herbicides
© 2022 The Authors. Published by Elsevier B.V.
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