title: Structure- and sequence-analysis inspired engineering of proteins for enhanced thermostability.
abstract: Protein engineering strategies for increasing stability can be improved by replacing random mutagenesis and high-throughput screening by approaches that include bioinformatics and computational design. Mutations can be focused on regions in the structure that are most flexible and involved in the early steps of thermal unfolding. Sequence analysis can often predict the position and nature of stabilizing mutations, and may allow the reconstruction of thermostable ancestral sequences. Various computational tools make it possible to design stabilizing features, such as hydrophobic clusters and surface charges. Different methods for designing chimeric enzymes can also support the engineering of more stable proteins without the need of high-throughput screening.
authors.affiliation: Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors.forename: Hein J
authors.initials: HJ
authors.lastname: Wijma
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Floor
authors.forename: Dick B
authors.initials: DB
authors.lastname: Janssen
mesh_terms: D000595:Amino Acid Sequence; D006358:Hot Temperature / Q000009:adverse effects*; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D017510:Protein Folding; D055550:Protein Stability; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D011506:Proteins