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@ARTICLE{Cui:904475,
      author       = {Cui, Haiyang and Pramanik, Subrata and Jaeger, Karl-Erich
                      and Davari, Mehdi D. and Schwaneberg, Ulrich},
      title        = {{C}ompass{R}-guided recombination unlocks design principles
                      to stabilize lipases in {IL}s with minimal experimental
                      efforts},
      journal      = {Green chemistry},
      volume       = {23},
      number       = {9},
      issn         = {1463-9262},
      address      = {Cambridge},
      publisher    = {RSC},
      reportid     = {FZJ-2021-06045},
      pages        = {3474 - 3486},
      year         = {2021},
      abstract     = {Biocatalysis in ionic liquids (ILs) has gained enormous
                      attention for the production of biodiesel, sugar esters, and
                      pharmaceuticals. However, hydrophilic IL interaction with
                      enzymes often results in reduced activity or even
                      inactivation. In this report, we prove that intrinsic lipase
                      stability and preservation of hydration shells of Bacillus
                      subtilis lipase A (BSLA) are two synergistic design
                      principles to retain enzymatic activity in ILs. After in
                      silico screening of nine beneficial amino acid positions by
                      the CompassR rule (in total, 172 variants), we rationally
                      designed two variants, to be constructed by site-directed
                      mutagenesis, and three libraries by site-saturation
                      mutagenesis. With minimal experiment effort, we identified
                      three all-around variants towards four [BMIM]-based ILs.
                      Remarkably, the variant M1a F17S/V54K/D64N/D91E/G155N had
                      6.7-fold higher resistance against $40\%$ (v/v) [BMIM]Cl,
                      5.6-fold in $80\%$ (v/v) [BMIM]Br, 5.0-fold in $30\%$ (v/v)
                      [BMIM][TfO], and 2.7-fold in $10\%$ (v/v) [BMIM]I compared
                      to wild-type BSLA, respectively, while showing 1.9-fold
                      improvement in specific activity. Computational analysis of
                      molecular dynamics and thermodynamic stability analysis of
                      the variants revealed the molecular basis for the resistant
                      variants M1a and M1b as the synergistic enhancement of
                      protein stability (ΔΔGfold ranging from −4.26 to −4.80
                      kcal mol−1) and increased hydration shells around the
                      substitutions in the four ILs (up to 1.7-fold). These design
                      principles and the gained molecular knowledge not only open
                      the door to direct experimentalists for rationally designing
                      promising IL-resistant enzymes, but also provide new
                      insights into enzymatic catalysis in ILs.},
      cin          = {IBG-1 / IMET},
      ddc          = {540},
      cid          = {I:(DE-Juel1)IBG-1-20101118 / I:(DE-Juel1)IMET-20090612},
      pnm          = {2172 - Utilization of renewable carbon and energy sources
                      and engineering of ecosystem functions (POF4-217)},
      pid          = {G:(DE-HGF)POF4-2172},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000644497200001},
      doi          = {10.1039/D1GC00763G},
      url          = {https://juser.fz-juelich.de/record/904475},
}