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Process for the preparation of cholesterol esterase
| Details |
Inventors: Beaucamp, Klaus; Nelboeck, Michael; Gauhl, Helmgard; Seidel, Hans; Gruber, Wolfgang; Brunner, Herwig;
Assignee: Boehringer Mannheim GmbH (Mannheim, DE)
Primary Examiner: Shapiro; Lionel M.
Assistant Examiner:
Attorney, Agent or Firm: Felfe & Lynch
The present invention provides a process for obtaining cholesterol esterase from micro-organisms of the genus Pseudomonas, wherein Pseudomonas sp. DSM 1280 or DSM 1281 is cultured in an appropriate culture medium in the presence of an inducer and the enzyme is obtained from the culture medium and/or from the cells. |
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DETAILED DESCRIPTION What is claimed is: 1.
Process for obtaining cholesterol esterase from micro-organisms, which process comprises culturing Psuedomonas sp.
DSM 1280 or DSM 1281 in a culture medium in the presence of an inducer, and recovering the enzyme obtained from the culture medium or the cells.
2.
Process as claimed in claim 1 wherein culturing is carried out in the presence of about 0.
1 to 5% by weight of inducer.
3.
Process as claimed in claim 1 wherein 0.
4 to 2% by weight of phosphate is added to the culture medium.
4.
Process as claimed in claim 1 wherein the micro-organisms Psuedomonas sp.
is DSM 1280.
5.
Process as claimed in claim 1 wherein the micro-organism Pseudomonas sp.
is DSM 1281.
Description:
The present invention is concerned with a process for obtaining cholesterol esterase from micro-organisms.
Cholesterol esterase has played an important role in clinical and biochemical analysis ever since processes have been developed for the enzymatic determination of cholesterol.
Since a large part of the cholesterol in biological material is present in the form of esters, the joint use of cholesterol esterase and cholesterol-oxidizing enzymes, such as cholesterol oxidase or cholesterol dehydrogenase, makes possible a completely enzymatic determination of cholesterol esters.
This is known from Federal Republic of Germany Patent Specification No.
2,264,847.
The enzyme from micro-organisms has proved to be especially suitable for determination processes involving the use of cholesterol esterase (see Federal Republic of Germany Patent Specification No.
2,506,712).
However, a disadvantage of the previously discovered micro-organisms with a content of cholesterol esterase which is sufficiently great to make a working up thereof worthwhile is the relatively low yields of enzyme activity which are obtained.
Surprisingly, we have now found that when using certain micro-organisms, several times higher activities can be achieved than was previously possible.
Thus, according to the present invention, there is provided a process for obtaining cholesterol esterase from micro-organisms of the genus Pseudomonas, wherein Pseudomonas sp
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Molecular Biology 
