title: Emerging technologies for protease engineering: New tools to clear out disease.
abstract: Proteases regulate many biological processes through their ability to activate or inactive their target substrates. Because proteases catalytically turnover proteins and peptides, they present unique opportunities for use in biotechnological and therapeutic applications. However, many proteases are capable of cleaving multiple physiological substrates. Therefore their activity, expression, and localization are tightly controlled to prevent unwanted proteolysis. Currently, the use of protease therapeutics has been limited to a handful of proteases with narrow substrate specificities, which naturally limits their toxicity. Wider application of proteases is contingent upon the development of methods for engineering protease selectivity, activity, and stability. Recent advances in the development of high-throughput, bacterial and yeast-based methods for protease redesign have yielded protease variants with novel specificities, reduced toxicity, and increased resistance to inhibitors. Here, we highlight new tools for protease engineering, including methods suitable for the redesign of human secreted proteases, and future opportunities to exploit the catalytic activity of proteases for therapeutic benefit. Biotechnol. Bioeng. 2017;114: 33-38. © 2016 Wiley Periodicals, Inc.
authors: Department of Chemical Engineering, University of California, Santa Barbara, California 93106.
authors: Jennifer L
authors: JL
authors: Guerrero
authors: Department of Chemical Engineering, University of California, Santa Barbara, California 93106.
authors: Patrick S
authors: PS
authors: Daugherty
authors: Department of Chemical Engineering, University of California, Santa Barbara, California 93106.
authors: Michelle A
authors: MA
authors: O'Malley
mesh_terms: D000818:Animals; D002460:Cell Line; D004926:Escherichia coli; D057166:High-Throughput Screening Assays; D006801:Humans; D010447:Peptide Hydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000627:therapeutic use; D015003:Yeasts
chemical_list: D011994:Recombinant Proteins; D010447:Peptide Hydrolases
keywords: cell-based assay; directed evolution; enzymes; high-throughput screening; proteases