Fig. 4: Validation of the DaArgC variants. | Nature Communications

Fig. 4: Validation of the DaArgC variants.

From: Engineering a new-to-nature cascade for phosphate-dependent formate to formaldehyde conversion in vitro and in vivo

Fig. 4

a Crystal structure of DaArgC (PBD-ID 8AFU, 2.0 Å) and b DaArgC3 (PBD-ID 8AFV, 2.2 Å). NADPH (gray) is modeled based on the structure of Mycobacterium tuberculosis ArgC (PBD-ID 2I3G). Active site residues affected by mutagenesis are shown as sticks. c In vivo productivity of the DaArgC variants. EcAckA and DaArgC variants were produced in E. coli BL21 DE3 ΔfrmRAB ΔpflAB and formate was added to the supernatant. The rate of formaldehyde production was evaluated by linear regression of four time points. Mean with standard deviation and individual data points are shown. Data points represent independent biological replicates (n = 3). d In vitro formaldehyde production of EcAckA + DaArgC variants. Data was evaluated as in c. Mean with SD and individual data points are shown. Data points represent independent technical replicates (n = 3). EcAckA Escherichia coli acetate kinase, DaArgC Denitrovibrio acetiphilus N-acetyl-γ-glutamyl phosphate dehydrogenase. Line plots of c and d are shown in Supplementary Fig. 9. Source data are provided as a Source Data file.

Back to article page