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@ARTICLE{Nutschel:872624,
      author       = {Nutschel, Christina and Fulton, Alexander and Zimmermann,
                      Olav and Schwaneberg, Ulrich and Jaeger, Karl-Erich and
                      Gohlke, Holger},
      title        = {{S}ystematically scrutinizing the impact of substitution
                      sites on thermostability and detergent tolerance for
                      {B}acillus subtilis lipase {A}},
      journal      = {Journal of chemical information and modeling},
      volume       = {60},
      number       = {3},
      issn         = {1549-960X},
      address      = {Washington, DC},
      publisher    = {American Chemical Society64160},
      reportid     = {FZJ-2020-00116},
      pages        = {1568-1584},
      year         = {2020},
      abstract     = {Improving an enzyme’s (thermo )stability or tolerance
                      against solvents and detergents is highly relevant in
                      protein engineering and biotechnology. Recent developments
                      have tended towards data-driven approaches, where available
                      knowledge about the protein is used to identify substitution
                      sites with high potential to yield protein variants with
                      improved stability and, subsequently, substitutions are
                      engineered by site directed or site saturation (SSM)
                      mutagenesis. However, the development and validation of
                      algorithms for data-driven approaches has been hampered by
                      the lack of availability of large-scale data measured in a
                      uniform way and being unbiased with respect to substitution
                      types and locations. Here, we extend our knowledge on
                      guidelines for protein engineering following a data-driven
                      approach by scrutinizing the impact of substitution sites on
                      thermostability or / and detergent tolerance for Bacillus
                      subtilis lipase A (BsLipA) at very large-scale. We
                      systematically analyze a complete experimental SSM library
                      of BsLipA containing all 3439 possible single variants,
                      which was evaluated as to thermostability and tolerances
                      against four detergents under respectively uniform
                      conditions. Our results provide systematic and unbiased
                      reference data at unprecedented scale for a
                      biotechnologically important protein, identify consistently
                      defined hot spot types for evaluating the performance of
                      data-driven protein engineering approaches, and show that
                      the rigidity theory and ensemble-based approach Constraint
                      Network Analysis yields (CNA) hot spot predictions with an
                      up to 9-fold gain in precision over random classification.},
      cin          = {JSC / NIC / ICS-6 / IMET},
      ddc          = {540},
      cid          = {I:(DE-Juel1)JSC-20090406 / I:(DE-Juel1)NIC-20090406 /
                      I:(DE-Juel1)ICS-6-20110106 / I:(DE-Juel1)IMET-20090612},
      pnm          = {511 - Computational Science and Mathematical Methods
                      (POF3-511) / Forschergruppe Gohlke $(hkf7_20170501)$ / PhD
                      no Grant - Doktorand ohne besondere Förderung
                      (PHD-NO-GRANT-20170405)},
      pid          = {G:(DE-HGF)POF3-511 / $G:(DE-Juel1)hkf7_20170501$ /
                      G:(DE-Juel1)PHD-NO-GRANT-20170405},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:31905288},
      UT           = {WOS:000526390800046},
      doi          = {10.1021/acs.jcim.9b00954},
      url          = {https://juser.fz-juelich.de/record/872624},
}