Single Residue Mutations and Characterization of Small Laccase (SLAC) from Streptomyces coelicolor

Students:

  • Victoria Maziarz
  • Arie Van Wieren
  • Alyssa Chesnutwood
  • Tori Swartz
  • Garrett Hoffman
  • Kaitlyn Adams

Abstract:

The group completed three different single residue mutations within SLAC: Ala189Arg, Val290Leu, and Glu228Asp. Small Laccase is an enzyme from a larger class of laccase enzymes which are copper oxidoreductases, and they are found in bacteria, fungi, and plants. SLAC can be made easily by E. coli in large quantities using recombinant biotechnology for use in various applications. SLAC is a potential valuable industrial enzyme involved in the production of biofuels and other cellulose based chemicals. Last semester in Biochemistry Lab 1 [BIOC 311], we cloned, expressed, purified, and characterized wild-type SLAC. This semester, in Biochemistry Lab 2 [BIOC 412], we used different computational methods to explore SLAC structure. Three residues were selected for mutation. These mutants were generated, and the corresponding mutant proteins were expressed and purified.  Finally, those purified mutant proteins will be characterized using enzyme kinetics.

Poster:

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