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US 2006/0188888 A1Published application
Method of screening for improved specific activity of enzymes
AssigneeNovozymes A/S
InventorShiro Fukuyama
Life of this patent
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Abstract
This invention relates to a method for screening libraries of enzyme variants for changes in specific activity by expression of a fusion protein consisting of at least two enzymes. By using one enzyme as a marker changes in specific activity for the other enzyme can be screened efficiently.
Claims17
1-16. (canceled)
A method of screening enzymes for variants with improved specific activity, comprising the steps of
- (i) generating a library of nucleic acid sequences encoding enzyme variants of interest
- (ii) providing a nucleic acid sequence encoding an enzyme to be fused with the enzyme in (i)
- (iii) fusing nucleic acid sequence encoding enzyme variants in (i) with nucleic acid sequence encoding enzyme in (ii)
- (iv) transforming the fused nucleic acid sequence obtained in (iii) into a host cell
- (v) culturing host cell in (iv) in order to express the fused enzymes
- (vi) sampling each cell culture obtained in (v)
- (vii) analyzing samples obtained in (vi) by determining activity ratio of the expressed fused enzymes
- (viii) selecting the samples exhibiting the desired activity ratio.
5 more elements
The method of claim 1, wherein the fused enzymes in 1(v) are an extracellular product.
Dependencies are inferred from the claim text.