Abstract
DNA family shuffling of 26 protease genes was used to create a library of chimeric proteases that was screened for four distinct enzymatic properties. Multiple clones were identified that were significantly improved over any of the parental enzymes for each individual property. Family shuffling, also known as molecular breeding, efficiently created all of the combinations of parental properties, producing a great diversity of property combinations in the progeny enzymes. Thus, molecular breeding, like classical breeding, is a powerful tool for recombining existing diversity to tailor biological systems for multiple functional parameters.
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Acknowledgements
We thank Dr. Titur Kretzschmar for initial amplification of the subtilisin fragments; Andreas Crameri, Steven delCardayre, and Claus Krebber for technical advice; and Sun Ai Raillard for stimulating discussions and comments on the manuscript.
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Ness, J., Welch, M., Giver, L. et al. DNA shuffling of subgenomic sequences of subtilisin. Nat Biotechnol 17, 893–896 (1999). https://doi.org/10.1038/12884
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DOI: https://doi.org/10.1038/12884
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