GBF Gesellschaft für Biotechnologische Forschung mbH, Braunschweig
Abstract
A lipase from a species of Pseudomonas has been cloned and expressed
in E. coli. Variants of this lipase have been generated, using sitespecific
mutagenesis, that have significantly altered k,,,, K, and
substrate specificity. We have undertaken to determine the threedimensional
structure of this enzyme using X-ray crystallography.
Crystals have been obtained and these crystals diffract to 2.5 Angstrom
resolution. We are now in the process of evaluating heavy atom
derivatives to be used to improve the phase information used to calculate
electron density maps
