Classifications

• • C—CHEMISTRY; METALLURGY
  • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
  • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
  • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
  • C11D3/16—Organic compounds
  • C11D3/38—Products with no well-defined composition, e.g. natural products
  • C11D3/386—Preparations containing enzymes, e.g. protease or amylase
  • C11D3/38663—Stabilised liquid enzyme compositions
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
  • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates

Definitions

• the invention relates to a liquid enzyme concentrate which can be used as a raw material solution for the production of liquid detergents or the like.
• enzymes in particular proteases
• the enzymes are processed into solid granular preparations for use in solid detergents.
• liquid detergents advance, there is also a need for such preparations to be able to produce storable stock solutions which can then be added to the liquid detergent in a subsequent processing step.
• Enzyme solutions of hydrolases, especially proteases are not per se stable in storage. This applies in particular to those enzyme solutions which are to be used for liquid detergents, since in the purification of the enzyme solutions apparently, in addition to impurities, substances which cause stabilization are also removed.
• a liquid detergent is proposed in which a polyol is used together with boric acid to stabilize the enzyme.
• the ratio polyol: boric acid should be less than 1, i.e. Boric acid is used compared to the polyol in excess.
• German Offenlegungsschrift DE 3125533 describes an aqueous builder-containing liquid detergent with enzymes, detergent substances, a polyol and boric acid or with salts thereof in a polyol: boric acid weight ratio of more than 1, with part of the crosslinked polyacrylate also being present.
• 1,2-propanediol and, in addition to other boron compounds also boric acid and its salts are mentioned.
• the amount of the polyol should be 100 to 200% by weight of the amount of boric acid. It should preferably be 100 to 160% by weight of the amount of boric acid.
• the hydrolase content of liquid detergents is usually only around 0.1% by weight of protein, at best up to 0.5% by weight. No conclusions can be drawn from the aforementioned German patent application as to how a stable enzyme liquid concentrate can be obtained. It is therefore an object of the invention to provide such a liquid concentrate.
• the invention relates to a liquid enzyme concentrate based on a hydrolase solution obtained from the cell mass and at least partially freed from soluble admixtures and having an enzyme content of 30,000 to 400,000 hydrolase units, comprising:
• boric acid and / or its soluble salts 0.5-5% by weight boric acid and / or its soluble salts and, if desired:
• the enzyme liquid concentrates according to the invention are concentrated, obtained by fermentation and at least partially purified enzyme solutions which, apart from the stabilizing agents used according to the invention and the substances present or formed during fermentation, contain no further constituents.
• a combination of propylene glycol and boric acid and / or their salts is used as stabilizing agent.
• the amount of propylene glycol (1,2-propanediol) is 20 to 40% by weight, in particular 20 to 30% by weight.
• the amount of boric acid is 0.5 to 5% by weight, in particular 1 to 3% by weight.
• the percentages by weight relating to boric acid relate to the acid, even if its salts are present in the solution and consequently higher amounts by weight are present.
• the soluble alkali metal or ammonium salts of the corresponding carboxylic acids can be used as salts of 1 to 3 basic carboxylic acids with up to 6 carbon atoms. Salts of formic acid, propionic acid, malonic acid, maleic acid or fumaric acid can also be used here. Usual concentrations of the salts based on the total preparation are between 0.5 and 5% by weight. For example, favorable results were obtained with 1% by weight of formic acid or 1% by weight of maleic acid, each used as the sodium salt.
• boric acid As a salt, i.e. adjust the enzyme solutions to a pH between 6 and 8 before, after or during the addition of boric acid.
• Suitable boric acid salts are the alkali metal or ammonium salts, especially the sodium or potassium salt.
• enzyme solutions can be used as the starting enzyme solution, such as are produced in the fermentation of yeast or bacteria, which are capable of forming hydrolases.
• Solutions which contain hydrolases, proteases, in particular subtilisin proteases, are preferred. Among them, those based on Subtilisin Carlsberg or Subtilisin BBN 1 are preferred.
• the cell fractions are separated from the fermenter solutions and then the enzymes are enriched. This can be carried out, for example, by a combination of microfiltration and ultrafiltration or else by precipitation and flocculation steps with, if desired, subsequent evaporation of the aqueous solution. 1,2-propanediol and boric acid are added to the purified enzyme solutions thus obtained.
• 1,2-propanediol and boric acid are added to the purified enzyme solutions thus obtained.
• the desired pH between 6 and 8 can be adjusted with sodium hydroxide solution before or during the addition of the boric acid.
• the preparations according to the invention can also contain preservatives. In many cases, this is not necessary because the combination of propylene glycol and boric acid has a preservative effect.
• a suitable preservative is, for example, formaldehyde.
• the invention is based on the synergistic stabilizing effect of boric acid and propylene glycol on such enzymes, which provides particularly favorable stability values within the specified limits, as can be seen in the examples below.
• subtilisin Carlsberg protease with an activity of 130 EP units / g was incubated with amounts of 0, 20, 30 and 50% by weight of propylene glycol and subjected to an accelerated aging test at 60 ° C. The following residual activities were measured after 24 hours:
• the enzyme activity was measured in all cases by the so-called EPE method, described in the German patent application DE 37 34047.
• the measuring principle is the cleavage of a phenolic ester bond of N-CBZ-amino acid p-nitrophenyl ester at 25 ° C. and a pH of 6.0, the concentration of the resulting p-nitrophenol being determined photometrically at 340 nm.

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• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Organic Chemistry (AREA)
• Engineering & Computer Science (AREA)
• Health & Medical Sciences (AREA)
• Wood Science & Technology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Genetics & Genomics (AREA)
• Zoology (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Oil, Petroleum & Natural Gas (AREA)
• Biomedical Technology (AREA)
• Biotechnology (AREA)
• Molecular Biology (AREA)
• Microbiology (AREA)
• Medicinal Chemistry (AREA)
• Biochemistry (AREA)
• General Engineering & Computer Science (AREA)
• General Health & Medical Sciences (AREA)
• Detergent Compositions (AREA)
• Enzymes And Modification Thereof (AREA)

Applications Claiming Priority (2)

Publications (1)

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• 1989
  • 1989-06-08 DE DE3918761A patent/DE3918761C1/de not_active Expired - Fee Related
• 1990
  • 1990-05-30 EP EP90110248A patent/EP0401661A1/de active Pending
  • 1990-05-30 CA CA002060514A patent/CA2060514A1/en not_active Abandoned
  • 1990-05-30 EP EP90908505A patent/EP0477210A1/de not_active Withdrawn
  • 1990-05-30 FI FI915671A patent/FI915671A0/fi not_active Application Discontinuation
  • 1990-05-30 JP JP2507952A patent/JPH04505851A/ja active Pending
  • 1990-05-30 WO PCT/EP1990/000863 patent/WO1990015129A1/de not_active Ceased
  • 1990-06-07 PT PT94299A patent/PT94299A/pt not_active Application Discontinuation

Non-Patent Citations (1)

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