Polypeptides Having DNase Activity
a polypeptide and dnase activity technology, applied in the direction of detergent compositions, bacteria, fungi, etc., can solve the problems of textile greasiness, malodor trapped in the organic structure, and organic matter such as biofilm is not desirable on textiles and surfaces, so as to maintain or improve the whiteness of the item
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example 1
nd Expression of Bacterial DNases
[0327]The DNases were derived from bacterial strains isolated from environmental samples by standard microbiological isolation techniques. Isolated pure strains were identified and taxonomy was assigned based on DNA sequencing of the 16S ribosomal genes (Table 1). The strains Saccharothrix australiensis (DSM43800) and Kutzneria albida (DSM43870) were purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig, Germany)
TABLE 1StrainSource CountryMature protein SEQ ID:Streptomyces sp-63712United States3Saccharothrix australiensisAustralia6Kutzneria albidaJapan9
[0328]Chromosomal DNA was isolated from pure cultures of the individual strains with the DNeasy Blood & Tissue Kit from Qiagen (Hilden, Germany) and subjected to full genome sequencing using Illumina technology. Genome sequencing, the subsequent assembly of reads and the gene discovery (i.e., annotation of gene functions) are known to the person skilled in the art an...
example 2
ion of Recombinant Enzymes by Nickel Affinity Chromatography
[0330]The pH of the cleared supernatant was adjusted to pH 8, filtrated through a 0.2 μM filter, and the supernatant applied to a 5 ml HisTrap™ excel column. Prior to loading, the column had been equilibrated in 5 column volumes (CV) of 50 mM Tris / HCl pH 8. In order to remove unbound material, the column was washed with 8 CV of 50 mM Tris / HCl pH 8, and elution of the target was obtained with 50 mM HEPES pH 7+10 mM imidazole. The eluted protein was desalted on a HiPrep™ 26 / 10 desalting column, equilibrated using 3 CV of 50 mM HEPES pH 7+100 mM NaCl. This buffer was also used for elution of the target, and the flow rate was 10 ml / min. Relevant fractions were selected and pooled based on the chromatogram and SDS-PAGE analysis.
example 3
Preparation of Biofilm Swatches
[0331]Biofilm swatches were made by growing Brevundimonas sp. on polyester swatches for two days. The biofilm swatches were rinsed twice in water and dried for 1 hour under a flow and subsequently punched into small circles and stored at 4° C. for further use.
Wash Experiment
[0332]Biofilm swatch punctures were placed in a deep well 96 format plate. The 96 well plate was placed in a Hamilton robot and subjected to a wash simulation program using the following conditions: Shaking speed: 30 sec at 1000 rpm. Duration of wash cycle: 30 minutes with shaking; temperature 30° C.; Volume of wash liquor (total): 0.5 ml per well. (490 wash liquor+10 uL sample).
[0333]Model detergent A wash liquor (100%) was prepared by dissolving 3.33 g / l of model detergent A containing 12% LAS, 11% AEO Biosoft N25-7 (NI), 5% AEOS (SLES), 6% MPG (mono propylene glycol), 3% ethanol, 3% TEA (triethanolamine), 2.75% cocoa soap, 2.75% soya soap, 2% glycerol, 2% sodium hydroxide, 2% sod...
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