Żądło-Dobrowolska Anna, Hammerer Lucas, Pavkov-Keller Tea, Gruber Karl, Kroutil Wolfgang
Institute of Chemistry, University of Graz, NAWI Graz, BioTechMed Graz, Heinrichstrasse 28, 8010 Graz, Austria.
ACIB GmbH, Petersgasse 14, 8010 Graz, Austria.
ACS Catal. 2020 Jan 17;10(2):1094-1101. doi: 10.1021/acscatal.9b04617. Epub 2019 Dec 27.
The biocatalytic Friedel-Crafts acylation has been identified recently for the acetylation of resorcinol using activated acetic acid esters for the synthesis of acetophenone derivatives catalyzed by an acyltransferase. Because the wild-type enzyme is limited to acetic and propionic derivatives as the substrate, variants were designed to extend the substrate scope of this enzyme. By rational protein engineering, the key residue in the active site was identified which can be replaced to allow binding of bulkier acyl moieties. The single-point variant F148V enabled the transformation of previously inaccessible medium chain length alkyl and alkoxyalkyl carboxylic esters as donor substrates with up to 99% conversion and up to >99% isolated yield.
最近已确定生物催化的傅克酰基化反应可用于间苯二酚的乙酰化反应，该反应使用活性乙酸酯，由酰基转移酶催化合成苯乙酮衍生物。由于野生型酶仅限于乙酸和丙酸衍生物作为底物，因此设计了变体以扩展该酶的底物范围。通过合理的蛋白质工程，确定了活性位点中的关键残基，该残基可以被取代以允许结合更大的酰基部分。单点变体F148V能够将以前无法使用的中链长度烷基和烷氧基烷基羧酸酯作为供体底物进行转化，转化率高达99%，分离产率高达>99%。
