Dataset related to the immobilization of the enzyme formate dehydrogenase (FDH) on different IMAC supports, one-step purification / co-immobilization of FDH with the enzyme glycerol dehydrogenase (GlyDH), study of GlyDH inhibition by DHA, adsorption study on the co-immobilized biocatalyst and the immobilization support, multi-enzymatic synthesis of formate and DHA using pure and crude substrates, and performance metrics of the biocatalyst in both reactions.

METHODOLOGICAL INFORMATION

1. Description of the methods used to collect and generate the data:

The enzymatic activity of the enzymes was measured spectrophotometrically at 340 nm using a phosphate buffer, NAD+ at 1.67 and 5 mM, 100 mM sodium formate, 100 mM glycerol, and a variable amount of soluble enzyme. The protein concentration was quantified spectrophotometrically at 595 nm using the Bradford method. The concentration of soluble CO2 in the medium was measured using a CO2 sensor (InPro5000i/12/220, Mettler Toledo S.A.E). Formate, DHA and glycerol quantification were performed in the liquid chromatograph using an ion exchange method with the IC-Sep COREGEL 87H3 column and sulfuric acid (H2SO4) 0.5 mM: Acetonitrile (65:35) as mobile phase. A flow rate of 0.6 mL·min-1, injection volume of 20 µL, column temperature of 30 °C, UV/Visible detector at 210 nm and RID detector at 30 °C. The co-immobilization of the enzymes was carried out in 100 mM phosphate buffer + 100 mM NaCl at pH 7.5, at a temperature of 4 °C, with a glutaraldehyde coating for GlyDH at 0.05% v/v for 150 minutes. The inhibition study was performed by adding different concentrations of DHA to the GlyDH activity assay. The adsorption isotherms of each compound were obtained by incubating 1 gram of the adsorbent with 9 mL of different concentrations in 100 mM phosphate buffer at pH 7.0, with agitation at 1200 rpm, at 30 °C, and for an indefinite period. The multi-enzymatic synthesis of formate and DHA was carried out in a stirred-tank reactor using 100 mM phosphate buffer at pH 7.5, 100 mM glycerol, 1 mM NADH, with CO₂ continuously bubbled at 1 VVM, and 20 grams of biocatalyst in a final volume of 200 mL. The reaction was stirred at 300 rpm, maintained at 30 °C, and run for 85 hours.

1. Data processing methods:

2. All data was processed using Excel.

3. Software or instruments needed to interpret the data:

4. Excel

5. Information about instruments, calibration and standards: Varian Cary 50 Bio UV-visible Spectrophotometer (Agilent) SPECTROstar Nano (BMG LABTECH) Spectrophotometer Fast Protein Liquid Chromatography (FPLC) in ÄKTA Pure (GE Healthcare®, Chicago, IL, USA) Liquid chromatograph Agilent 1220 Infinity II Multi Therm Heat Block system (Benchmark Scientific Inc.). SpinChem reactor (SpinChem, Sweeden)

6. Environmental or experimental conditions: The reactions were carried out in a fume hood at 30°C and within a pH range of 7.0 to 8.5.