DE4226162A1 - STABLE ENZYMPREPAIR - Google Patents

Abstract

A stable, aqueous enzyme preparation comprises cellulase(s) and protease(s), in a buffer of pH from 4 to 6 and comprises an oxygen-containing, water-soluble, organic solvent (preferably ethylene glycol, propylene glycol, glycerol or polyethylene glycol). The compositions may be utilised in the after-treatment of mixed fabrics.

Description

The invention relates to a new, stable enzyme preparation, which in addition Cellulase also contains protease, and its use to achieve of fashionable effects on blended fabrics consisting of silk or wool on the one hand and cotton, linen or viscose on the other hand and Treatment of raw wool.

Both cellulases and proteases are (separately) as aftertreatments known for textile materials, especially the cleaning Effect in the sense of the breakdown of cellulose - resp. Protein residues in the front reason stood. For some years now cellulases have been increasingly used used cotton to achieve fashionable effects and Proteases are also used as an aid for the felt-free finishing of wool known.

Even if cellulase and protease are occasionally used together (Japanese Kokai 49/1 21 807, for example, describes their use in washing water from raw wool to isolate the wax), no expert would the enzyme domain remember a stable preparation that contains both cellulase as well as containing protease. On the contrary, due to the be knew that proteases also degrade other enzymes rather expect that the cellulases in such preparations will be destroyed. In the published PCT application WO 90/0 75 690 is actually emp foal, the cellulases used after treatment with Proteases (e.g. with a detergent containing such enzymes) to destroy.

It has now surprisingly been found that aqueous preparations which contain both cellulase (s) and protease (s) can, which are stable by at least 2-3 months permanent storage time only a slight loss of activity of both Show cellulase (s) as well as the protease (s).

The invention thus relates to stable aqueous preparations of Cellulase (s) - protease (s) mixtures, both with a buffer mixture are adjusted to pH 4-6, as well as an oxygen-containing water contain soluble organic solvent.

All enzymes or enzyme mixtures with the EC number can be used as cellulase 3.2.1.4 are used to attack cellulose by attacking β-glycosidic Break ties. Their activity is expressed in cellulase units (CU) presses, with one unit corresponding to the amount of enzyme listed under Standard conditions, d. H. at pH 5 and 40 ° C after 20 minutes of incubation the substrate carboxymethyl cellulose (type 7 LFD, Hercules), a lot of right producing carbohydrates corresponding to 1 pmol glucose per minute. Examples of such cellulases are the products bacterial or fungal Origin of

• - Aspergillus,
• - Neurospora,
• - Phizopus,
• - Trichoderma and
• - Trametes species.

Examples of such microorganisms are:

• - Humicola insolens,
• - Aspergillus oryzae,
• - Trichoderma viride (reesei),
• - Rhizopus sp.,
• - Humicola grisea,  
• - Penicillium spinulosum,
• - Trichurus cylindricus,
• - Clostridium thermocellum,
• - Corynebacterium sp.,
• - Sclerotium bataticola,
• - Pseudomonas fluoroescens var. Cellulosa
• - Bacterioides succinogenes
• - Trametes sanguinea.

Proteases which are used according to the enzyme classi certification (EC No.) according to the recommendations of the "Nomenclature Committee of the International Union of Biochemistry "(Enzyme Nomenclature, published for the ZUS by Academic Press, Inc., San Diego, California, USA 1984) der numerically belong to the large class of peptide hydrolases (EC 3.4).

The proteases can

• a) animal,
• b) vegetable,
• c) be of microbial origin,

for example

• a) Pancreatic proteases
  Trypsin
  Chymotrypsin
• b) plant proteases
  Papain
  Ficin
  Bromelain
• c) - from Bacillus species:
  B. subtilis
  B. licheniformis
  B. alkalophilus
  B. cereus
  B. natto
  B. vulgatus
  B. mycoides
  - from Aspergillus species:
  A. flavus oryzae
  A. niger
  A. saitoi
  A. usamii
  - from Mucor and Rhizopus species:
  Pusillus muscle
  M. rentrei

Enzymes of the serine proteinase type (EC 3.4.21) such as, for. B. Chimotrypsin, chimotrypsin C, metridium proteinase A, trypsin, thrombin, Plasmin, acrosine, Myxobacter alfa lytic proteinase, Alternaria serine proteinase, Tenebrio a proteinase and especially Microbial serine proteinase (EC 3.4.21.14). For the determination of the proteolytic enzyme activity Azocasein is used as the substrate, with the Azocasein Units (ACU) on Measured values based on the following standard conditions:

• - temperature 40 ° C,
• - pH 8,
• - incubation time 30 minutes,
• - spectrophotometric measurement at 340 nm,

(Modified method according to H.U. Bergmeyer, methods of enzymatic Analysis, Verlag Chemie Weinheim, FRG 1970, page 978 and ff.)

On the one hand, other components of the aqueous preparation are low molecular, oxygen-containing, water-soluble organic solvents, such as glycerol and / or ethylene glycol, propylene glycol-1,2, propylene glycol 1,3, ethylene carbonate, propylene carbonate, polyethylene glycols and others hand salts (buffer mixtures) with which the aqueous preparation on a pH value of 4-6, preferably 5, is set: as examples thinks: triethanolamine maleate, triethanolamine citrate, sodium citrate, Tris (hydroxymethyl) aminomethane maleate, glycyl-glycine-piperazine di-hydro chloride NaOH; Citric acid disodium hydrogen phosphate; Sodium acetate  Acetic acid; Succinic acid-NaOH; Potassium hydrogen phthalate NaOH; Potassium dihydrogenphosphate-di-sodium hydrogenphosphate.

The various components of the preparation can vary in quantity vary. Quantities are required for good stability and storage stability of 5-50% enzyme (as an aqueous solution of cellulase (s) and protease (s) in proportions from 8: 1 to 1: 6), from 5-45% glycerin or ethylene glycol, Propylene glycol 1,2, propylene glycol 1,3, ethylene carbonate, propylene carbon nat, polyethylene glycols and from 0.1 to 5% salt recommended, all Percentages refer to the weight of the whole preparation and Water makes up the missing percentages. 25-45% enzyme are preferred (in a ratio of 1: 1), 20-40% solvent and 0.5-2.5% salt ent hold.

These preparations are used for the aftertreatment of mixed fabrics made of silk or wool on the one hand and cotton, linen or viscose on the other on the other hand. These textile materials receive a be after the treatment particularly soft handle and the fashionable appearance of the so-called "sand wash" -Effect. You can also treat raw wool to remove the vegetable residues and the vegetable fiber components are used and show a very good carbonation effect.

This aftertreatment is preferably carried out using the exhaust method in a tumbler, the enzyme preparation in an amount of 0.5 to 20 g / l, preferably from 4 to 8 g / l and in a pH range from 4 to 11 preferably 5 to 10 is used at a liquor ratio of 1:20. A particularly preferred method is to treat the starting at pH 5-6 and ending at pH 9-10. The Temperature of the treatment bath is based on the optimal activity of the enzymes set and is usually 40-60 ° C. Instead of in a tumbler you can the treatment is also carried out in the J-box, reel runner, jet or overflow leads.

In the following examples, parts are by weight and Percentages by weight. The temperatures are in degrees Celsius specified.

MANUFACTURING EXAMPLES Preparation 1

They are produced by mixing the following Components according to their weight proportions in the order:

62.5 parts water
2.5 parts of tris (hydroxymethyl) aminomethane maleate
10 parts cellulase (EC 3.2.1.4) in aqueous solution containing 4500 CU / g
15 parts of protease of the serine type (EC 3.4.21) in aqueous solution containing 3100 ACU / g
10 parts glycerin.

This preparation with a pH of 5 is at least 3 months in the room temperature stable in storage. The loss of activity in both Protease as well as with cellulase during this storage period a maximum of 10%.

Preparation 2

The procedure is as described in Example 1 and mixing

28.0 parts water
2.0 parts of triethanolamine maleate
25 parts of cellulase (as in Example 1)
25 parts of protease (as in Example 1)
20 parts of 1,2-propylene glycol.

Preparation 3

The procedure is as described in Example 1 and mixing

63.5 parts water
1.5 parts of tris (hydroxymethyl) aminomethane maleate
10 parts cellulase (as in Ex. 1)
10 parts protease (as in Ex. 1)
15 parts of ethylene glycol.

Preparation 4

The procedure is as described in Example 1 and mixing

12.5 parts water
2.5 parts of triethanolamine citrate
40 parts of cellulase (as in Example 1)
5 parts protease (as in Ex. 1)
40 parts propylene glycol 1,2

Preparation 5

The procedure is as described in Example 1 and mixing

31 parts of water
4 parts of triethanolamine maleate
5 parts cellulase (as in Example 1)
30 parts of protease (as in Ex. 1)
30 parts propylene glycol 1,2

Preparation 6

The procedure is as described in Example 1 and mixing

19.5 parts water
0.5 parts tris (hydroxymethyl) aminomethane maleate
10 parts cellulase (as in Ex. 1)
30 parts of protease (as in Ex. 1)
40 parts glycerin

Preparation 7

The procedure is as described in Example 1 and mixing

65 parts of water
5 parts Na citrate
5 parts cellulase (as in Example 1)
20 parts protease (as in Ex. 1)
5 parts of polyethylene glycol MG 300

APPLICATION EXAMPLES

A. A dyed wool / cotton blend is in a liquor 1:20, which contains 6 g of preparation 1 in water of 10 ° dH per liter, first treated for 15 minutes at a pH of 5 to 6 and 50 °. The pH is then adjusted to 9 to 10 with 3 g of soda calc./l and treated for a further 15 minutes at 50 °. The treatment is done in a tumbler. After treatment, it will Washed tissue in 40 ° warm water and cold water and dried. The wool-cotton fabric gets a soft feel and the fashionable look of the sand-wash effect.

Instead of the wool / cotton blend, fibers or fabrics can also be used consisting of

• - wool / viscose,  
• - wool / linen,
• - silk / viscose,
• - silk / cotton,
• - silk / linen,

be treated.

B. A dyed silk-cotton blend becomes 1:10 in a fleet with 4 g / l of preparation 5 at pH 5 and 60 ° C for 1 hour treated. Then adjust the pH with soda calc. to pH 9-10 and treat the tissue for another 20 minutes at 60 °. After draining the bath, the mixed fabric becomes warm once and once washed cold. The fabric gets a soft feel and the fashionable Appearance of the sand-wash effect.

C. A dyed silk-cotton blend is treated in a 1:20 liquor with the following ingredients:
1 g / l of a commercially available wetting agent,
6 g / l of preparation 6.
The bath is heated to 60 ° and held at this temperature for 20 minutes; The fabric is washed and dried as described in Example 1.
The fabric gets a soft feel and the appearance of the sand-wash effect.

D. Raw wool is treated in a 1:20 fleet with the following components:
2 g / l of a commercially available wetting agent,
6 g / l of preparation 2.
The bath is heated to 60 ° C and held at this temperature for 1 hour.
After draining the bath, the wool is washed warm and cold. The vegetable residues and vegetable fiber parts adhering to the raw wool are optimally removed. There is no damage to the wool fiber.

E. Example D is repeated, using instead of preparation 2 Preparation 5 is used.

F. In a modification of example D, the fleet is after one hour Treatment of wool at 60 ° with soda calc. adjusted to pH 9-10. The wool is treated for a further 30 minutes at this pH and 60 °. The carbonation effect is very good.

Claims (5)

1. Stable, aqueous preparations of cellulase (s) and protease (s), the with a buffer mixture and an oxygen-containing, water-soluble organic solvents are adjusted to a pH 4-6. 2. Preparations according to claim 1 which are adjusted to pH 5. Stable, aqueous preparations of cellulase (s) and protease (s) according to claim 1 or 2, the ethylene glycol, propylene glycol, glycerin or contain polyethylene glycol. 3. Stable preparations according to one of claims 1 to 3, characterized characterized in that they contain 5 to 50% enzyme (as an aqueous solution of Cellulase (s) and protease (s) in the ratio of 8: 1 to 1: 6), 5 to Contain 45% solvent and 0.1 to 5% buffer salt in addition to water. 4. Process for post-treatment of blended fabrics consisting of silk or wool on the one hand, and cotton, linen or viscose on the other hand, characterized in that aqueous preparations of Cellulase (s) and protease (s) according to any one of claims 1 to 4 be set. 5. Process for the treatment of raw wool, characterized in that aqueous preparations of cellulase (s) and protease (s) according to one of claims 1 to 4 are used.