Immobilization of Bacillus subtilis Oxalate Decarboxylase on a Zn-IMAC Resin

Umar Twahir,a Laura Molina,a Andrew Ozarowski,b and Alexander Angerhofer a*

a Department of Chemistry, University of Florida, Gainesville, Florida 32611-7200, USA.b National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310, USA

Supplemental Information

Figure S1: Comparison of Michaelis-Menten kinetic analysis of decarboxylase activity of free OxDC at pH 4.2 (royal blue) and pH 5.5 (red) as well as IMAC-bound OxDC at pH 4.2 (green) and pH 5.5 (cyan), IMAC-bound OxDC at pH 4.2 after 10 freeze-thaw cycles (orange).

Figure S2: The 406.4 GHz HF-EPR at 3K spectra of WT-OxDC free enzyme pH 4.06 (red), immobilized pH 3.88 (black) and simulation based on site A (blue) previously determined magnetic parameters shown in table S1. Spectral Parameters: 50 kHz modulation frequency, 25 G modulation amplitude, 2 mT/s sweep rate.

Figure S3: The 406.4 GHz HF-EPR at 20K spectra of WT-OxDC free enzyme (left panel), immobilized (right panel) and simulation based on site H (blue) previously determined magnetic parameters shown in table S1. Spectral Parameters: 50 kHz modulation frequency, 25 G modulation amplitude, 2 mT/s sweep rate.

Figure S4: Free and Immobilized WT-OxDC and the effects of freezing and thawing the resin bound enzyme. Respective concentrations of immobilize enzyme reflected in table 2. Spectral Parameters: 100 kHz modulation frequency, 10 G modulation amplitude, 0.63 mW microwave power, and temperature set to 5 K.

Figure S5: X-band EPR at 5K comparing IMAC resin loaded with 50 mM free MnCl2. Spectral Parameters: 100 kHz modulation frequency, 10 G modulation amplitude, 0.63 mW microwave power, and temperature set to 5 K.