Synthetic Biology - Parts - Protein Engineering

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Researchers

Jennifer Anthony, Jeffrey Dietrich, Matt Garcia, Yasuo Yoshikuni

Project Description

Structure
Structure

Protein engineering has been widely adapted as an efficient design methodology for protein functions. It is generally classified into two distinctive methodologies; rational design and directed evolution. In rational design, amino acid residues predicted to be important for particular protein functions based on the crystal structure and/or primary sequence analyses are altered. Then, the effects over the protein functions are analyzed. Due to the relative complexity of proteins and difficulty of prediction, the rational design approach is rarely successful. On the other hand, directed evolution is more reliable methodology to design protein functions. In directed evolution, mutations are randomly introduced into proteins, and mutant library is created. This library is then subjected to the high-through put (HTP) screening to isolate mutants with desired properties. However, directed evolution also often fails to design ideal protein functions, because the methodology is highly dependent on the efficiency of HTP screening. In Keasling lab, we developed new methodology called systematic remodeling to design protein functions without need for HTP screening, and applied the methodology to promiscuous terpene synthase -humulene synthase (one of the enzymes involved in terpenoid biosynthetic pathway: Figure) to create seven different variants each catalyzing very specific and different reaction mechanism (Figure). Currently, we are applying the developed methodology to other proteins.

Relevant Publications

  • Y. Yoshikuni and J. D. Keasling. 2007, "Pathway engineering by designed divergent evolution." Current Opinion in Chemical Biology. 11(2):233-239
  • Y. Yoshikuni, T. E. Ferrin, and J. D. Keasling. 2006. “Designed divergent evolution of enzyme function.” Nature 440:1078-1082.
  • Y. Yoshikuni, V. J. J. Martin, T. E. Ferrin, and J. D. Keasling. 2006. “Engineering cotton (+)-δ-cadinene synthase to an altered function: germacrene D-4-ol synthase.” Chemistry and Biology. 13:91-98.

Funding Resources

  • Bill and Melinda Gates Foundation


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