Conversion of Bacillus thermocatenulatus lipase into an efficient phospholipase with increased activity towards long-chain fatty acyl substrates by directed evolution and rational design |
| |
| Authors: | Kauffmann I; Schmidt-Dannert C |
| |
| Affiliation: | 1 Institute for Technical Biochemistry, University of Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany and
2 Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, 1479 Gortner Ave, St Paul, MN 55108, USA |
| |
| Abstract: | The thermoalkalophilic lipase from Bacillus thermocatenulatusBTL2 exhibits a low phospholipase activity (lecithin/tributyrinratio 0.03). A single round of random mutagenesis of the BTL2gene followed by screening of 6000 transformants on egg-yolkplates identified three variants with 10–12-fold increasedphospholipase activities, corresponding to lecithin/tributyrinratios of 0.16–0.36. All variants were specific for thesn-1 acyl ester bond of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine.Mutations occurred predominantly in the N-terminal part of BTL2with regions surrounding the predicted helix |
| |
| Keywords: | |
| 本文献已被 Oxford 等数据库收录！ |
|
