Premium
Insight into the Structure and Activity of Surface‐Engineered Lipase Biofluids
Author(s) -
Zhou Ye,
Jones Nykola C.,
Nedergaard Pedersen Jannik,
Pérez Bianca,
Vrønning Hoffmann Søren,
Vang Petersen Steen,
Skov Pedersen Jan,
Perriman Adam,
Kristensen Peter,
Gao Renjun,
Guo Zheng
Publication year - 2019
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201800819
Subject(s) - lipase , protein engineering , bacillus subtilis , chemistry , directed evolution , mutagenesis , enzyme , combinatorial chemistry , biochemistry , biology , mutation , mutant , gene , genetics , bacteria
Despite a successful application of solvent‐free liquid protein (biofluids) concept to a number of commercial enzymes, the technical advantages of enzyme biofluids as hyperthermal stable biocatalysts cannot be fully utilized as up to 90–99% of native activities are lost when enzymes were made into biofluids. With a two‐step strategy (site‐directed mutagenesis and synthesis of variant biofluids) on Bacillus subtilis lipase A (BsLA), we elucidated a strong dependency of structure and activity on the number and distribution of polymer surfactant binding sites on BsLA surface. Here, it is demonstrated that improved BsLA variants can be engineered via site‐mutagenesis by a rational design, either with enhanced activity in aqueous solution in native form, or with improved physical property and increased activity in solvent‐free system in the form of a protein liquid. This work answered some fundamental questions about the surface characteristics for construction of biofluids, useful for identifying new strategies for developing advantageous biocatalysts.