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Metalloprotease from kribbella aluminosa and detergent compositions comprising the metalloprotease

A composition and detergent technology, applied in the directions of detergent compositions, detergent compounding agents, biochemical equipment and methods, etc., can solve the problems of instability and non-commercialization of metalloprotease

Inactive Publication Date: 2017-02-15
NOVOZYMES AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] However, known metalloproteases are very unstable under normal wash conditions and in conventional detergent compositions
Therefore, the use of metalloproteases in detergents and in washing and cleaning processes is not currently commercialized

Method used

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  • Metalloprotease from kribbella aluminosa and detergent compositions comprising the metalloprotease
  • Metalloprotease from kribbella aluminosa and detergent compositions comprising the metalloprotease
  • Metalloprotease from kribbella aluminosa and detergent compositions comprising the metalloprotease

Examples

Experimental program
Comparison scheme
Effect test

example

[0355] Materials and Methods

[0356] Test material for washing determination

[0357] Grass stain swatch 062KC was obtained from Warwick Equest Ltd (Unit 55, Consett Business Park, Consett, County Durham, DH8 6BN, UK )acquired,

[0358] The remaining small swatches were obtained from the Center For Testmaterials BV (PO Box 120, 3133 KT Vlaardingen, The Netherlands).

[0359]

[0360] standard detergent

[0361] Standard detergents B, J, M and Y were used. Standard Detergent B is similar to a European type liquid detergent composition. Standard Detergent J is similar to an American type liquid detergent composition. Standard Detergent M is similar to a Chinese type powdered detergent composition. Standard Detergent Y is similar to the developing and emerging (D&E) Asia Pacific market type powder detergent composition.

[0362]

[0363]

[0364]

[0365]

[0366]

[0367]

[0368] washing assay

[0369] Automated Mechanical Stress Assay (AMSA) f...

example 1

[0404] Example 1. Cloning and expression of M4 metalloprotease from Alum korean bacteria

[0405] The metalloprotease was derived from a strain of Alumenum korea, which was isolated from a soil sample collected from China in 2009.

[0406] Chromosomal DNA from pure cultures was purified and subjected to whole genome sequencing using Illumina technology. The assembled genome sequence and subsequent analysis of the 16S ribosomal subunit gene sequence indicated correct taxonomic identification as a species of S. alumensis.

[0407] Genomic sequences were analyzed for metalloproteases from the MEROPS family M4 compared to the M4 protease NprE from Bacillus subtilis 168 (Uniprot P68736) by searching using the BLAST program. This analysis identified a gene encoding a putative M4 metalloprotease having the nucleotide sequence given in SEQ ID:1.

[0408] The gene encoding the M4 metalloprotease was amplified by PCR and fused with regulatory elements and homology regions for recombin...

example 2

[0413] Example 2. SDS-PAGE and N-terminal sequencing

[0414] When the purified protease preparation was analyzed by SDS-PAGE and the gel was stained with Coomassie, three bands at approximately 55 kDa, 47 kDa and 8 kDa were seen. Edman degradation, complete MS, and in-gel peptide map analysis showing that all three bands are derived from the M4 protease, indicating that bands 47kDa and 8kDa represent nicked molecules in which the full-length M4 protease cuts once in the loop And the two peptide chains are still associated with each other and give a fully active molecule. The relative intensities of bands 47kDa and 8kDa suggest a 1:1 molar ratio between these two bands, supporting this hypothesis.

[0415] N-terminal sequencing of the purified protein was performed and the mature enzyme was found to be initiated at GTGNSMY, confirming the identity of the enzyme and the presence of the N-terminal propeptide. Intact molecular weight analysis of purified proteins by mass spectr...

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Abstract

The present invention relates to a new metalloprotease and the use thereof in cleaning processes, such as laundry and dish wash. The invention also relates to detergent compositions and cleaning compositions comprising a metalloprotease.

Description

[0001] References to Sequence Listings [0002] This application contains a Sequence Listing in computer readable form. This computer readable form is hereby incorporated by reference. [0003] field of invention [0004] The present invention relates to a new metalloprotease (E.C 3.4.24) from Sangongra alumensis and its use in cleaning processes such as dishwashing and laundry washing. The present invention further relates to cleaning and / or detergent compositions comprising metalloproteases. [0005] Background of the invention [0006] For more than 30 years, the detergent industry has employed different enzymes in detergent formulations, the most commonly used enzymes include proteases, amylases and lipases, each suitable for removing different types of stains. In addition to these enzymes, detergent compositions typically comprise a complex combination of ingredients. For example, most cleaning products include surfactant systems, bleach or builders. [0007] Protease...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/52C12N15/57C11D3/386
CPCC11D3/386C12N9/52C12Y304/24
Inventor M.杰曼森S.尼曼德-格拉鲁普P.R.厄斯特加德J.比耶尔
Owner NOVOZYMES AS