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Improving Alternate Lignin Catabolite Utilization Of Ligab From Sphingobium Sp. Strain Syk-6 Through Site Directed Mutagenesis, Kevin P. Barry, Erin F. Cohn, Abraham Ngu, Erika A. Taylor

Erika A. Taylor, Ph.D.

Protocatechuate 4,5-dioxygenase (LigAB) catalyzes dioxygenation of multiple lignin derived aromatic compounds—such as protocatechuate (PCA), gallate (GA) and 3-O-methyl gallate (3OMG)—with decreasing proficiency as the molecule size increases. We predicted that phenylalanine-103 of the α subunit (Phe103α) controls substrate specificity through interaction with the C5-funtionality of bound substrates, and mutagenesis would enhance GA and 3OMG catalysis. LigAB with Phe103α mutations (F103 V, F103T and F103H) displayed enhanced catalytic efficiency for dioxygenation of 3OMG, with mutants displaying 12- to 31-fold increases in View the MathML source, making these mutant enzymes more active with 3OMG than its native dioxygenase (DesZ). The F103T and …