[Zurück]

T. Wiesinger, T. Bayer, S. Milker, M.D. Mihovilovic, F. Rudroff:
"Cell Factory Design and Optimization for the Stereoselective Synthesis of Polyhydroxylated Compounds";
ChemBioChem, 18 (2017), S. 1 - 9.

A synthetic cascade for the transformation of primary alcohols into polyhydroxylated compounds in Escherichia coli, through the in situ preparation of cytotoxic aldehyde intermediates and subsequent aldolase-mediated C C bond formation, has been investigated. An enzymatic toolbox consisting of alcohol dehydrogenase AlkJ from Pseudomonas putida and the di-hydroxyacetone-/hydroxyacetone-accepting aldolase variant Fsa1-A129S was applied. Pathway optimization was performed at the genetic and process levels. Three different arrangements of the alkJ and fsa1-A129S genes in operon, monocistronic, and pseudo-operon configuration were tested. The last of these proved to be most beneficial with regard to bacterial growth and protein expression levels. The optimized whole-cell catalyst, combined with a refined solid-phase extraction downstream purification protocol, provides diastereomerically pure carbohydrate derivatives that can be isolated in up to 91 % yield over two reaction steps.

alcohols · biocatalysis · domino reactions · enzymes · solid-phase extraction

http://dx.doi.org/10.1002/cbic.201700464

http://publik.tuwien.ac.at/files/publik_266583.pdf