Computer-aid screening of thermostable lipase LipA from Burkholderia sp. ZYB002

Abstract:

[Objective] We improved the thermostability of lipase LipA from Burkholderia cecapia ZYB002 using protein engineering technology.[Methods] Lipase LipA mutant library was designed and screened using the following software, YASARA, FoldX, Rosetta, and Gromacs. We screened 27 thermostable lipase LipA mutants displaying the salt bridge effect among the resulting library of 341 variants, and then further screened using the site-directed mutagenesis technology.[Results] Three mutants LipA-Asn¹²⁵Asp, LipA-Asn¹²⁵Glu and LipA-Gln²⁶²Glu displayed improved thermostability. The T₅₀¹² value of LipA-Asn¹²⁵Asp, LipA-Asn¹²⁵Glu and LipA-Gln²⁶²Glu increased by 4.0℃, 5.5℃ and 4.4℃, respectively. The half-life of LipA-Asn¹²⁵Asp, LipA-Asn¹²⁵Glu and LipA-Gln²⁶²Glu at 55℃ increased by 2.23-fold, 3.8-fold and 2.6-fold, respectively.[Conclusion] It is feasible to screen thermostable mutant from the computationally designed library.

Key words:Burkholderia sp.ZYB002;lipase LipA;in silico design and screening;salt bridge;thermostable mutant