Abstract
Previously, we evolved Lipase A from Bacillus subtilis towards increased thermostability. The resulting mutant retains significant catalytic activity upon heating above 60 °C (and up to 100 °C) and cooling down, whereas wild-type lipase precipitates irreversibly and does not show significant activity in these conditions. Kinetic thermostability of proteins has not been characterized well on the molecular structure level so far, therefore our aim is to study it using NMR spectroscopy. Here, nearly complete 1H, 13C and 15N resonance assignments are reported for wild-type and mutant Lipase A. Chemical shifts were used to predict secondary structure elements of both Lipase A variants.
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Acknowledgments
The NMR research was supported by the European Communities 6th Framework Programme Research Infrastructure Action program (project EU-NMR, contract # RII3-026145); W.A. was Marie Curie Outgoing International Fellow sponsored by the European Communities. Prof. Wiktor Koźminski and Jan Stanek (Faculty of Chemistry, Warsaw University) are acknowledged for their help in acquisition and processing of non-uniformly sampled NMR spectrum.
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Augustyniak, W., Wienk, H., Boelens, R. et al. 1H, 13C and 15N resonance assignments of wild-type Bacillus subtilis Lipase A and its mutant evolved towards thermostability. Biomol NMR Assign 7, 249–252 (2013). https://doi.org/10.1007/s12104-012-9420-z
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DOI: https://doi.org/10.1007/s12104-012-9420-z