Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1995-09-27
2000-05-23
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435196, 435136, 435155, C12N 916
Patent
active
06066486&
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a method of hydrolysing a cholesterol ester. More specifically, the invention relates to use of a cholesterol esterase enzyme obtainable from a strain of Pseudomonas fragi.
BACKGROUND ART
It is well known that some of the members belonging to the genus Pseudomonas possess the ability to produce cholesterol esterase, other members possess the ability to produce lipase, and that furthermore some members possess the ability to produce both types of enzymes.
In U.S. Pat. No. 4,283,494 a lipase derived from Pseudomonas alcaligenes, able to become activated by bile salts, and having cholesterol esterase activity is described.
In a study by Smirnov et. al. [Smirnov, V. V.; Kornyushenko, O. N.; Bioko, O. I.; Kolesova, E. A.; Govseeva, N. N.; Endt, V. P.; and Kiprianova, E. A.; Mikrobiologicheskii Zhurnal (Kiev); 42 (5), 1980, 566-570], a total of 591 strains of 25 species of bacteria belonging to the genus Pseudomonas were studied for their ability to synthesize extracellular cholesterol esterase. Only individual strains of Ps. aeruginosa, Ps. pseudoacaligenes, Ps. fluorescens, Ps. putida, and Ps. maltophilia (3.4% of the studied cultures) had cholesterol esterase activity. Lipolytic activity was found in Ps. aureofaciens (in 100% of the studied strains), Ps. cepacia (in 75%), Ps. maltophilia (in 60%), Ps. fluorescens (in 13.3%), and Ps. aeruginosa (in 60%).
From this study it is noticed that none of the Ps. fragi cultures examined were able to produce cholesterol esterases.
SUMMARY OF THE INVENTION
According to the present invention it has now been found that strains of Pseudomonas fragi are able to produce extracellular cholesterol esterase. Moreover, it has surprisingly been found that the enzyme component responsible for the cholesterol esterase activity is also the component responsible for the lipase activity.
Accordingly, the invention relates to the use of a cholesterol esterase obtainable from a strain of Ps. fragi, i.e. to a method of hydrolysing a cholesterol ester by treating the ester with said cholesterol esterase enzyme. The method of the invention is particular useful for the treatment of eggs, in processes for hydrolysis of resin in pulp, and in processes for hydrolysis or synthesis of sterols or lanolin.
In another aspect, the invention provides a cleaning composition comprising a cholesterol esterase obtainable from a strain of Ps. fragi.
BRIEF DESCRIPTION OF THE DRAWING
The present invention is further illustrated by reference to the accompanying drawing, in which:
FIG. 1 shows the kinetic curves (absorbance versus time) for various enzymes as obtained by spectrophotometric determination, cf. Example 2.
DETAILED DISCLOSURE OF THE INVENTION
Surprisingly, it has now been found that strains of Pseudomonas fragi possess the ability to produce enzymes having cholesterol esterase activity and that the enzyme responsible for the cholesterol esterase activity is identical to the enzyme responsible for the lipase activity, i.e. the very same protein possesses lipase activity as well as cholesterol esterase activity.
Accordingly, the present invention relates to a method of hydrolysing a cholesterol ester comprising treating the cholesterol ester with a cholesterol esterase enzyme obtainable from a strain of Pseudomonas fragi.
In a preferred embodiment, the cholesterol esterase is Lipase B from Ps. fragi, available from WAKO Pure Chemical Industries, Ltd., Japan, order No. 126-02931, and derived from the strain Ps. fragi 22.39B, FERM BP-1051, or a mutant or a variant thereof.
In a more preferred embodiment, the cholesterol esterase has the N-terminal amino acid sequence identified by SEQ ID No: 1 of the sequence listing attached to this specification.
Thus it should be noted that the enzyme used in the method of the invention having cholesterol esterase activity as well as lipase activity may be described e.g. as a cholesterol esterase acting lipase or a lipase acting cholesterol esterase.
The cholesterol esterase used in the method of the inv
REFERENCES:
Chemical Abstract--Abstract No. 3955, Wakako et al., Biochim. Biophys. Acta. (1991).
Chemical Abstract--Abstract No. 216788, Sumitomo Chemical Co. (1993).
Chemical Abstract--Abstract No. 36573, Buchert, et al., Appl. Microbiol. Biotechnol. (1988).
WPIDS--WPIDS Accession No. 78-60989A, Toyobo (1978).
Gregg, Esq. Valeta
Lankford , Jr. Leon B.
Novo Nordisk A S
Zelson Esq. Steve T.
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