A combination of mutational and computational scanning guides the design of an artificial ligand-binding controlled lipase.

ABSTRACT: Allostery, i.e. the control of enzyme activity by a small molecule at a location distant from the enzyme's active site, represents a mechanism essential for sustaining life. The rational design of allostery is a non-trivial task but can be achieved by fusion of a sensory domain, which responds to environmental stimuli with a change in its structure. Hereby, the site of domain fusion is difficult to predict. We here explore the possibility to rationally engineer allostery into the naturally not allosterically regulated Bacillus subtilis lipase A, by fusion of the citrate-binding sensor-domain of the CitA sensory-kinase of Klebsiella pneumoniae. The site of domain fusion was rationally determined based on whole-protein site-saturation mutagenesis data, complemented by computational evolution

SUBMITTER: Kaschner M 

PROVIDER: S-EPMC5316958 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature