Creating a more robust 5-hydroxymethylfurfural oxidase by combining computational predictions with a novel effective library design

Additional file 4: Figure S1. Michaelis–Menten graph of WT and 7xHMFO. Kinetic assay performed with HRP peroxidase in 50 mM phosphate buffer pH 8.0 at 25 °C using HMF as substrate

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Additional file 7: Table S5. Percentages of products formed during the oxidation of 5 mM HMF by 2 μM of enzyme in phosphate buffer 50 mM pH 8.0 at 40 °C in Eppendorf ThermoMixer C while shaking at 1000 rpm. Average values of two experiments (standard deviations were < 7%, with an average standard deviation of 1.0%). Samples with only phosphate buffer, substrate, and WT enzyme where used as control

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Additional file 9: Gene Sequences. Synthetic gene sequences for WT and 8xHMFO for the Golden Gate gene shuffling

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Additional file 8: Figure S3. Michaelis–Menten graph of 8BxHMFO. Kinetic assay performed with HRP peroxidase in 50 mM phosphate buffer pH 8.0 at 25 °C using FFA as substrate