Additional file 5. HPLC chromatograms of enantiomeric separations of reaction products. a Application of AtPrR2; b application of FiPLR. [3a] = (+)-pinoresinol 3a, [3b] = (−)-pinoresinol 3b, [4a] = (+)-lariciresinol 4a, [4b] = (−)-lariciresinol 4b, [5a] = (−)-secoisolariciresinol 5a

Esther Ricklefs

Marco Girhard

Vlada B. Urlacher

Microbial Cell Factories

Springer - Publisher Connector

Three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (−)-pinoresinol via kinetic resolution

Additional file 2. Formation of lariciresinol 4 or secoisolariciresinol 5 from pinoresinol 3 in the reactions with cleared cell lysates of different E. coli strains expressing the reductase AtPrR2 (a) or the reductase FiPLR (b). Reaction conditions: 50 mM KPi-buffer, pH 7.5, 0.2 mM pinoresinol 3, 50 µl cleared cell lysate containing AtPrR2 or FiPLR, 0.2 mM NADPH. Optionally, 3 U ml−1 GDH and 20 mM D-glucose were added for cofactor regeneration (cr). The reactions were incubated at 25 °C for 16 h. Besides formation of lariciresinol 4 or secoisolariciresinol 5, no other reaction products were observed

Esther Ricklefs (3400889)

Marco Girhard (3400892)

Vlada Urlacher (3400886)

The Francis Crick Institute

MOESM2 of Three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (−)-pinoresinol via kinetic resolution

Additional file 3. Cell growth of E. coli C41(DE3) cells co-transformed with pACYCtac_psvao and pET16b_cgl1 in the presence of different concentrations of eugenol 1. Cell cultures were grown at 37 °C, 180 rpm to an OD600 of 0.6 and harvested (indicated by an arrow). Cell pellet was resuspended in TB medium, eugenol 1 was added and the cultures were incubated at 37 °C, 180 rpm. Cell growth was monitored at 600 nm. Black square: 0 mM eugenol 1, blue circle: 1 mM, green triangle: 2.5 mM, purple square: 5 mM, orange circle: 10 mM

MOESM3 of Three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (−)-pinoresinol via kinetic resolution

Additional file 1. DNA-sequences of the synthetic genes (syatprr2 and syfiplr), plasmids and strains used within this study

MOESM1 of Three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (â)-pinoresinol via kinetic resolution

MOESM5 of Three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (−)-pinoresinol via kinetic resolution

https://figshare.com/articles/MOESM3_of_Three-steps_in_one-pot_whole-cell_biocatalytic_synthesis_of_enantiopure_-_and_-pinoresinol_via_kinetic_resolution/4318013

Three-steps in one-pot: whole-cell biocatalytic synthesis of enantiopure (+)- and (−)-pinoresinol via kinetic resolution

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The Francis Crick Institute

The Francis Crick Institute

Springer - Publisher Connector

The Francis Crick Institute