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Amino acids V12 (putative oxyanion hole), F112 (active site architecture), N108 (flavonoid binding) and C13 (GSH stabilization) were fully randomized by site saturation mutagenesis (NNK codons), and the created Pt GSTF8 variants were tested for conversion of flavan-3,3,4-triol ( 4 ) to cyanidin ( 8 ), and for their GSH-transferase activity towards CDNB ( 5 ) as a control of catalytic competence

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Anthocyanin production in yeast To highlight the practical relevance of our pathway elucidation, we set out to show that the catalytic activity of arGSTs is also required for anthocyanin biosynthesis in vivo in a eukaryotic cell and thus is key for production of anthocyanins ( 1 ) in microbial cell factories