Design and evolution of new catalytic activity with an existing protein scaffold

Hee Sung Park, Sung Hun Nam, Jin Kak Lee, Chang No Yoon, Bengt Mannervik, Stephen J. Benkovic, Hak Sung Kim

Research output: Contribution to journalArticle

193 Scopus citations

Abstract

The design of enzymes with new functions and properties has long been a goal in protein engineering. Here, we report a strategy to change the catalytic activity of an existing protein scaffold. This was achieved by simultaneous incorporation and adjustment of functional elements through insertion, deletion, and substitution of several active site loops, followed by point mutations to fine-tune the activity. Using this approach, we were able to introduce β-lactamase activity into the αβ/βα metallohydrolase scaffold of glyoxalase II. The resulting enzyme, evMBL8 (evolved metallo β-lactamase 8), completely lost its original activity and, instead, catalyzed the hydrolysis of cefotaxime with a (kcat/K m)app of 1.8 × 102 (mole/liter) -1 second-1, thus increasing resistance to Escherichia coli growth on cefotaxime by a factor of about 100.

Original languageEnglish (US)
Pages (from-to)535-538
Number of pages4
JournalScience
Volume311
Issue number5760
DOIs
StatePublished - Jan 27 2006

All Science Journal Classification (ASJC) codes

  • General

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    Park, H. S., Nam, S. H., Lee, J. K., Yoon, C. N., Mannervik, B., Benkovic, S. J., & Kim, H. S. (2006). Design and evolution of new catalytic activity with an existing protein scaffold. Science, 311(5760), 535-538. https://doi.org/10.1126/science.1118953