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Insight into the mechanism for enzymatic biofuel production

By Gerald Ondrey |

Enzymes that are responsible for the production of hydrocarbons in blue-green algae — a hydrocarbon producing cyanobacteria — have now been identified by the research group of Munehito Arai at The University of Tokyo (Japan; www.c.u-tokyo.ac.jp). Along with this achievement, the researchers discovered the important amino acids within the enzymes that enable the enzymes to work efficiently, and believe the breakthrough can be applied for the efficient production of renewable biodiesel fuels.

The researchers compared the amino-acid sequences and the hydrocarbon-producing activities of ten representative aldehyde-deformylating oxygenases (ADOs), which catalyze a difficult and unusual reaction in the conversion of aldehydes to hydrocarbons — a technique that has been widely used for biofuel production in metabolic engineering. The activity was found to be highest for the ADO from Synechococcus elongatus PCC 7942 (7942ADO). In contrast, the ADO from Gloeobacter violaceus PCC 7421 (7421ADO) had low activity but yielded high amounts of soluble protein, resulting in a high production level of hydrocarbons. By introducing 37 single amino acid substitutions at the non-conserved residues of the less active ADO (7421ADO) to make its sequence more similar to that of the highly active ADO (7942ADO), they found 20 mutations that improved the activity of 7421ADO. In addition, 13 other mutations increased the amount of soluble ADO while maintaining more than 80% of wild-type activity. Correlation analysis showed a solubility-activity trade-off in ADO, in which activity was negatively correlated with solubility.

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