Figure 7From: Dieselzymes: development of a stable and methanol tolerant lipase for biodiesel production by directed evolutionStructure of PML mutant Dieselzyme 4. (A) Cartoon representation of Dieselzyme 4. The locations of clustered mutations described in the text, designated Region 1 and Region 2, are highlighted with a red and orange box respectively. Mutations in Dieselzyme 4 are labeled and shown as sticks. Lid helices α5 and α8 are shown for clarity and colored green. The catalytic triad is shown as orange ball and stick. (B) 90° rotation of (A). (C) Zoom in of Region 1 as seen in the wild-type PML structure. (D) Zoom in of Region 1 as seen in the Dieselzyme 4 structure. Mutations are highlighted as magenta sticks. New H-bonds present in Dieselzyme 4 are shown as dashed black lines. (E) Zoom in of Region 2 as seen in the wild-type PML structure. The “helical loop” between residues 202 and 207 is highlighted orange. Potential H-bonds between residues R220 and H216 from helix α8, the loop region, and crystallographic waters are shown as orange dashes. The bound calcium is shown as a cyan sphere. (F) Zoom in of Region 2 as seen in the Dieselzyme 4 structure. The remodeled “helical loop” between residues 202 and 207 is highlighted in orange. Residues G202E and E207 that form potential new salt-bridges with R220 and/or H216 from helix α8 are highlighted with a star. Other H-bonds are shown as red dashes. The bound calcium is shown as a cyan sphere.Back to article page