Alkaline bacillus lipases, coding DNA sequences therefor and bac

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

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4352521, 536 232, C12N 920, C12N 1555, C12N 120

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active

054279364

DESCRIPTION:

BRIEF SUMMARY
The present invention relates to alkaline bacillus lipases and to DNA sequences, which code for these lipases, to the use and to a method for producing these lipases, as well as to the bacillus strains, which are capable of forming these lipases.
Enzymatic compositions for washing, cleaning and bleaching applications are well known in the art. Admittedly, various types of enzymes have already been proposed for these applications. However, the main interest has been directed essentially to the proteases and amylases. The lipases, previously proposed in the art for washing, cleaning and bleaching compositions, were obtained by culturing microorganisms such as pseudomonas, rhizopus, chromobacter and humicola species (including thermomyces species).
Although such lipases have been considered as possible enzymes for the applications cited, lipases have hardly gained any acceptance previously in washing, cleaning and bleaching compositions, since various constituents of these compositions have a negative effect on the activity of the lipases. For example, it is well known that synthetic anionic surfactants, in particular, have a negative effect on lipase activity. Admittedly, of the plurality of enzymes known in the art, which belong to the class of lipases, each individual enzyme also has special, advantageous properties. Nevertheless, the possibilities of employing these enzymes are limited because of their disadvantageous properties.
Moreover, the lipases, since they themselves also are proteins, are subject to the proteolytic degradation by detergent proteases, if they are used in combination with these proteases as detergent components of washing, cleaning and bleaching compositions. As a result, the danger exists that, when protease and lipase are used together in this composition, the lipolytic activity cannot be utilized at all or utilized only partly because of the lipolytic activity losses.
On the other hand, fats and oils are removed only poorly and incompletely from the fabrics and/or objects that are to be cleaned, particularly at low washing and cleaning temperatures, such as temperatures up to 40.degree. C., if the washing and cleaning process is not supported by lipolytically effective enzymes.
It is therefore an object of the invention to make available such lipases, which are useful as additives for detergent, cleaning agent and bleaching agent compositions and have a high activity at temperatures up to 40.degree. C. and a versatile applicability in the presence of proteases and over a wide pH range and particularly in a neutral to alkaline pH range.
Surprisingly, it has now been discovered that lipases, which are secreted by bacillus species, have the desired properties with a pH optimum in the alkaline pH range and a temperature optimum between about 30.degree. and 40.degree. C. In appropriate refinements of the invention, it is a question particularly of alkaline bacillus lipases, which can be obtained by culturing Bacillus pumilus.
In preferred embodiments of the invention, particularly those alkaline bacillus lipases of the aforementioned species are present, which have an amino acid sequence, which is at least 70%, preferably at least 80% and particularly at least 90% homologous with the amino acid sequence given in FIG. 1 (SEQ ID NO:2). Homology is understood here to be the degree of relationship of the amino acid sequence in question of a bacillus lipase to the amino acid sequences of the lipases from the natural bacillus isolates DSM 5776, DSM 5777 or DSM 5778 and, in particular to the amino acid sequence (SEQ ID NO:2) of the lipase from the natural bacillus isolate DSM 5776, as given in FIG. 1 (SEQ ID NOS:1 and 2). To determine the homology, in each case the sections of amino acid sequence of the lipases from the natural bacillus isolates, particularly the sections of the amino acid sequence of FIG. 1 (SEQ ID NO:2) and an amino acid sequence of a bacillus lipase, which is to be compared therewith, are superimposed, so that there is maximum agreement between the two amino acid sequence

REFERENCES:
patent: 5093256 (1992-07-01), Shen et al.
Chemical Abstract 188319d, vol. 94, No. 23, p. 301, (1981).
Chemical Abstract 130743g, vol. 113, No. 15, p. 521, (1990).

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