Cholesterol esterases with variable substrate specificity
AssigneeBoehringer Mannheim GmbH
InventorsGunther Schumacher, Christian Hanke, Stephan Fischer
Life of this patent
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Abstract
For the preparation of cholesterol esterases with variable substrate specificity by cloning of a cholesterol esterase gene, the active center of which has the sequence -Gly-His-Ser-X-Gly-, wherein X signifies an amino acid, into a vector, transformation of a micro-organism with this vector and expression of the cholesterol esterase gene, one exchanges the amino acid X of the active center for another amino acid by mutagenesis.
Claims13
Process for preparation of a cholesterol esterase, comprising mutagenizing a gene which codes for a first cholesterol esterase with an active site having amino acid sequence:
- Gly-His-Ser-X-Gly
- to change X to an amino acid which differs from the amino acid in said first cholesterol esterase, to form a gene which codes for a second cholesterol esterase, wherein said second cholesterol esterase has substrate specificity different from said first cholesterol esterase, transforming a microorganism with said gene, and culturing said microorganism to produce said cholesterol esterase with different substrate specificity, said mutagenizing said gene being such that X is changed to His.
Dependencies are inferred from the claim text.