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Savinase variants having an improved wash performance on egg stains

a technology of savinase and egg stains, which is applied in the direction of sugar derivatives, biochemistry apparatus and processes, enzymes, etc., can solve the problems of pronounced egg stains removal problems in e.g. laundry or hard surfaces, and achieve the effect of improving the wash performance on egg stains

Inactive Publication Date: 2009-08-27
NOVOZYMES AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about improving the ability of a subtilase enzyme to clean egg stains. This is achieved by modifying the parent enzyme, Savinase, by adding certain residues to its structure. The modifications result in a more effective cleaning performance on egg stains.

Problems solved by technology

In particular, the problem of removing egg stains from e.g. laundry or hard surfaces has been pronounced due to the fact that substances present in the egg white inhibit many serine proteases.

Method used

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  • Savinase variants having an improved wash performance on egg stains
  • Savinase variants having an improved wash performance on egg stains

Examples

Experimental program
Comparison scheme
Effect test

example 1

Construction and Expression of Subtilases According to the Invention

[0169]Example 1 covers both SEQ ID NO:2, and SEQ ID NO:4. It is to be understood that the term SEQ ID NO:2 at any time can be replaced by the term SEQ ID NO:4.

[0170]The subtilisin having the amino acid sequence shown in SEQ ID NO:2 was located in plasmid pKH400 (previously described in WO 98 / 41623). PKH400 was constructed from pJS3 (E. coli-B. subtilis shuttle vector containing a synthetic gene encoding for subtilase 309 (Savinase®) as described by J. Schiodt et al. in Protein and Peptide Letters, 3, 39-44 (1996)) by introduction of two BamHI sites at positions 1841 and 3730.

[0171]It includes an origin of replication for E. coli and Bacillus; the cat gene conferring resistance towards chloramphenicol, the promoter directing the initiation of transcription of the subtilisin and the pre / pro regions from Savinase®.

[0172]This plasmid replicates both in E. coli and in Bacillus subtilis, the subtilisins according to the i...

example 2

Automatic Mechanical Stress Assay (AMSA)

Description of AMSA-Test Method:

[0183]Washing experiments are performed in order to assess the wash performance of selected protease variants in detergent compositions. The proteases of the present application are tested using the Automatic Mechanical Stress Assay (AMSA). With the AMSA, the wash performance of a large quantity of small volume enzyme-detergent solutions can be examined. The AMSA plate has a number of slots for test solutions and a lid firmly squeezing the textile swatch to be washed against all the slot openings. During the washing time, the plate, test solutions, textile and lid are vigorously shaken to bring the test solution in contact with the textile and apply mechanical stress in a regular, periodic oscillating manner. For further description see WO 02 / 42740 especially the paragraph “Special method embodiments” at page 23-24.

[0184]The experiment was conducted under the experimental conditions specified below:

Commercial de...

example 3

Microtiter Egg Assay (MEA)

[0191]In this assay the digestion of denatured egg proteins by proteases in the presence of detergent was followed in a 96-well microtiter plate. Heating of egg proteins produces visual changes and changes in physicochemical properties. The clear translucent material is trans-formed to a milky substance. This is partly due to sulfhydryl-disulfide interchange reactions of denatured proteins. For example, heating unmasks the sulfhydryl group of ovalbumin, and the unmasked groups form disulfide linkages. The digestion of the denatured egg proteins by proteases converts the milky egg solution to a more clear solution dependent on the ability of the enzymes to degrade egg proteins.

[0192]Procedure

[0193]a) Make an egg solution of 200 mg egg powder (Sanovo International AS) and 93.7 mL 16° dH water. Denature the egg solution while the temperature increases to 85° C.;

[0194]b) Dilute the subtilase enzyme to 320 nM in succinic acid buffer: 10 mM succinic acid+2 mM CaC...

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Abstract

Subtilase variants having an improved wash performance on egg stains. These subtilases are useful exhibiting excellent or improved wash performance on egg stains when used in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.

Description

TECHNICAL FIELD[0001]The present invention relates to novel subtilases having an improved performance on soil, in particular egg stains. These subtilases are useful exhibiting excellent or improved performance on egg stains when used in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.[0002]The present invention also relates to isolated polynucleotides encoding the subtilases, nucleic acid constructs, recombinant expression vectors, host cells comprising the nucleic acid construct, and methods for producing and using the subtilases of the invention. Further, the present invention relates to cleaning and detergent compositions comprising the subtilase enzymes of the invention as well as to use of such enzymes in detergent compositions and for removal of egg stains.BACKGROUND OF THE INVENTION[0003]In the detergent industry enzymes have for more than 30 years been implemented in washin...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C11D3/386C12N9/54C12N15/11C12N15/00C12N1/21C12N1/19C12P21/02
CPCC11D3/386C11D3/38627C12N9/50C11D3/38645C11D3/38654C11D3/38636C12N9/54
Inventor MINNING, STEFANKNÖTZEL, JÜRGEN CARSTEN FRANZSØRENSEN, NIELS HENRIK
Owner NOVOZYMES AS
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