title: ¹H, ¹³C and ¹⁵N resonance assignments of wild-type Bacillus subtilis Lipase A and its mutant evolved towards thermostability.
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 (1)H, (13)C and (15)N 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.
authors.affiliation: Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim, Germany.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Augustyniak
authors.forename: Hans
authors.initials: H
authors.lastname: Wienk
authors.forename: Rolf
authors.initials: R
authors.lastname: Boelens
authors.forename: Manfred T
authors.initials: MT
authors.lastname: Reetz
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry; D002247:Carbon Isotopes; D004795:Enzyme Stability; D019143:Evolution, Molecular; D008049:Lipase / Q000737:chemistry*; D050505:Mutant Proteins / Q000737:chemistry*; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D011522:Protons; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D002247:Carbon Isotopes; D050505:Mutant Proteins; D009587:Nitrogen Isotopes; D011522:Protons; D008049:Lipase