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Improving Alternate Lignin Catabolite Utilization of LigAB from Sphingobium sp. strain SYK-6 through Site Directed Mutagenesis
Process Biochemistry (2015)
  • Kevin P Barry, Wesleyan University
  • Erin F Cohn, Wesleyan University
  • Abraham Ngu, Wesleyan University
  • Erika A Taylor, Wesleyan University
Abstract

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 F103 V point mutants also exhibited allosteric activation for the dioxygenation of PCA and GA, respectively, in the presence of vanillin, as previously observed for LigAB. The enhanced utilization of substrates by these mutants makes them potentially useful for efforts to develop engineered organisms that catabolize lignin into biofuels or fine chemicals.

Keywords
  • Protocatechuate 4,
  • 5-dioxygenase,
  • Dioxygenase,
  • Gallate,
  • 3-O-methyl gallate,
  • Lignin,
  • PCAD superfamily
Publication Date
July, 2015
Citation Information
Kevin P Barry, Erin F Cohn, Abraham Ngu and Erika A Taylor. "Improving Alternate Lignin Catabolite Utilization of LigAB from Sphingobium sp. strain SYK-6 through Site Directed Mutagenesis" Process Biochemistry Vol. in press (2015)
Available at: http://works.bepress.com/eataylor/1/