Subtilase variants

A technology of protease and variant, applied in the field of automatic dishwashing composition, producing and using the variant of the present invention, cleaning and detergent composition, which can solve the problems such as difficult to completely remove stains

Active Publication Date: 2012-08-22
NOVOZYMES AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Washing conditions are changing all the time, e.g. due to temperature and pH considerations, and many stains remain difficult to completely remove under normal washing conditions

Method used

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Experimental program
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preparation example Construction

[0138] Preparation of variants

[0139] Variants of the parent protease variants can be made according to any mutagenesis method known in the art, such as site-directed mutagenesis, synthetic gene construction, semi synthetic gene construction, random mutagenesis, shuffling (shuffling) etc. to prepare.

[0140] Site-directed mutagenesis is a technique for creating one or a few mutations at defined sites on a polynucleotide molecule encoding a parental protease variant. The technique can be performed in vitro or in vivo.

[0141] Synthetic gene construction involves the in vitro synthesis of polynucleotide molecules designed to encode a polypeptide molecule of interest. Gene synthesis can be performed using a variety of techniques, such as the multiplex microchip-based technique described by Tian et al. (Tian et al., Nature 432:1050-1054), and similar techniques in which oligonucleotides are synthesized acid and assembled on a photo-programmable microfluidic chip.

[0142] ...

Embodiment 2

[0357] Stain Remover Example 2: Typical European Powder Stain Remover Composition

[0358]

[0359]

[0360] The enzymes of the detergent composition according to the invention can use conventional stabilizers and protease inhibitors, for example polyols such as propylene glycol or glycerol, sugars or sugar alcohols, different salts such as NaCl; KCl; lactic acid, formic acid, boric acid or boric acid derivatives For example aromatic boronic acid esters, or phenylboronic acid derivatives such as 4-methylphenylboronic acid, or peptide aldehydes such as di-, tri- or tetrapeptide aldehydes or aldehyde analogs (forms B1-BO-R, where R is H, CH3, CX3, CHX2 or CH2X (X=halogen), B0 is a single amino acid residue (preferably with optionally substituted aliphatic or aromatic side chains); and B1 consists of one or more amino acid residues (preferably one, two or three), optionally containing N-terminal protecting groups, or as described in WO09118375, WO98 / 13459) or protein-type p...

Embodiment 1

[0393] Construction and expression of enzyme variants

[0394] Variants of the invention can be constructed and expressed by methods known to those skilled in the art. The following is a possible example of how to prepare a variant of the invention:

[0395] Site-directed mutagenesis:

[0396] Subtilisin 309 of the invention comprising specific insertions / deletions / substitutions Targeted variants are prepared by conventional cloning of PCR-generated DNA fragments (Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor, 1989) using oligos containing the desired mutations.

[0397] The template plasmid DNA may be pSX222, or an analog thereof containing a subtilisin 309 variant. Mutations were introduced into the variant constructions by oligo-directed mutagenesis.

[0398] The subtilisin 309 variant was transformed into E. coli. DNA purified from overnight cultures of these transformants was transformed into Bacillus subtilis by restrictio...

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Abstract

The present invention relates to protease variants.The present invention also relates to polynucleotides encoding the variant protease variants and to nucleic acid constructs, vectors, and host cells comprising the polynucleotides, and methods of using the variant enzymes, such as, in laundry and detergent compositions.

Description

[0001] Sequence listing involved [0002] This application contains a Sequence Listing in computer readable form. The computer readable form is incorporated herein by reference. technical field [0003] The present invention relates to novel subtilase variants which exhibit an alteration in one or more properties relative to the parent subtilase, including: wash performance, thermostability, storage stability or catalytic activity. The variants of the invention are suitable for example in cleaning or detergent compositions, such as laundry detergent compositions and dishwashing compositions, including automatic dishwashing compositions. The invention also relates to isolated DNA sequences encoding the variants, expression vectors, host cells, and methods for producing and using the variants of the invention. Furthermore, the invention relates to cleaning and detergent compositions comprising the variants according to the invention. Background technique [0004] In the det...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C11D3/386C12N9/54C12N15/09
CPCC11D3/386C12N9/54C12N15/52
Inventor J.C.F.诺泽尔M.N.霍考夫L.贝尔A.贝尼
Owner NOVOZYMES AS
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