Boehringer Mannheim Gmbh

Klaus Beaucamp

Wolfgang Gruber

Hans Seidel

Gunter Lang

Hans Ulrich Bergmeyer

Peter Roeschlau

Erich Bernt

Helmgard Gauhl

Herwig Brunner

Albert Roder

Helmut Lill

Michael Nelbock-Hochstetter

Michael Nelboeck

Detlef Von Hoerschelmann

Joachim Ziegenhorn

Ralf Mattes

Gunter Weimann

Knut Bartl

Klaus Muhlegger

Alexander Hagen

Hans Mollering

Peter Stahl

Walter Schneider

Dieter Jaworek

Michael Nelboeck-Hochstetter

Peter Buckel

Peter Wunderwald

Gunter Schumacher

Karl-Heinz Botsch

Georg Schawohl

Gerhard Berger

Armin Masuth

Johanna Gramsall

Gunter Holz

To prepare a microorganism producing .alpha.-galactosidase, not only a DNA containing an .alpha.-galactosidase gene but also a vector which is appropriate to the transformable cells to be used and contains antibiotic resistance genes are completely split with restriction endonuclease Sal I, the fragment of approximately four megadaltons of relative molecular weight is obtained from the fragments of the DNA containing the .alpha.-galactosidase gene, is mixed with the solution of the vector also split with Sal I, and is recombined in the presence of DNA ligase with the formation of a recombinant DNA. The recombinant DNA obtained is incubated with transformable cells with transformation of the recombinant DNA into the cells, the transformed cells are cultured on a nutrient substrate containing raffinose as sole carbon source, the antibiotic-resistant colonies formed are isolated and lysed, the plasmid DNA is isolated from the lysate, the plasmid DNA is split with restriction endonuclease Hind III or Eco R I, and the obtained solution is diluted, and treated with DNA ligase. The renatured plasmids obtained are again introduced into transformable cells, the transformed cells are again cultured on a nutrient substrate containing raffinose as carbon source as well as antibiotic, and the colonies formed which do not utilize raffinose are isolated. A micro-organism is thus obtained which does form an .alpha.-galactosidase requiring no cofactors, but which does not form invertase.

Process for the preparation of a microorganism which forms

The present invention provides a micro-organism of the species Escherichia coli or Pseudomonas putida, wherein it constitutively forms creatinamidinohydrolase. The present invention also provides a process for the production of such a micro-organism.

Microorganisms and plasmids for the constitutive formation of

The present invention provides a process for obtaining glucose dehydrogenase by culturing a micro-organism in an appropriate nutrient medium and obtaining the enzyme from the cell mass or from the culture medium, wherein the micro-organism cultured is Bacillus cereus DSM 1644. The nutrient medium used contains a carbon source which consists predominantly or solely of glycerol.

Process for obtaining glucose dehydrogenase and micro-organism therefor

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.

Process for the preparation of cholesterol esterase

The present invention provides a process for obtaining cholesterol esterase from micro-organisms, wherein a micro-organism capable of cholesterol esterase formation is cultured in an appropriate nutrient medium in the presence of lecithin as inducer and the enzyme is obtained from the culture medium and/or from the cells.

Process for obtaining cholesterol esterase from micro-organisms

The present invention provides a process for obtaining cholesterol oxidase, wherein Streptomyces griseofuscus DSM 40191, Streptomyces hygroscopicus 40771, Streptomyces acidomyceticus DSM 40798 and/or Arthrobacter paraffinens DSM 312 are cultured and the enzyme obtained from the culture supernatant and/or the cells.

Process for obtaining cholesterol oxidase

A control reagent for heparin activity determination comprising

Control reagent for heparin activity determination

Cholesterol is determined by incubating a sample suspected of containing cholesterol in an aqueous medium with cholesterol oxidase and determining either the oxygen consumption, the hydrogen peroxide formed, or cholestenone, as a measure of the initial cholesterol content; reagent compositions comprising cholesterol oxidase and a system for the determination of H.sub.2 O.sub.2 or for the determination of cholestenone are provided.

Reagent and method for the determination of cholesterol

A novel peroxidase for reduced nicotinamide-adenine-dinucleotide is provided which peroxidase oxidizes reduced nicotinamide-adenine-dinucleotide with hydrogen peroxide to give nicotinamide-adenine-dinucleotide and water and is characterized by a Michaelis constant K.sub.M

NADH Peroxidase for hydrogen peroxide determination

Cholesterol oxidase free of unstabilizing residual traces of detergent is activated by adding to the cholesterol oxidase, before use thereof, at least one surface-active compound with lipophilic and hydrophilic properties. Diagnostic agents for determining cholesterol comprising cholesterol oxidase, a hydrogen peroxide detection system, a buffer, and at least one surface-active compound having lipophilic and hydrophilic properties are provided.

Activation of cholesterol oxidase for cholesterol assay

A new enzyme, cholesteroloxidase, is prepared by recovering same from microorganisms, preferably by washing a cholesterol-converting microorganism with butanol/water, disintegrating the microorganism culture and extracting same with a buffer solution containing a non-ionic surface-active agent, and isolating cholesterol oxidase from the extract.

Cholesteroloxidase and method for its production from microorganisms

Enzymes, particularly labile enzymes, and most particularly microorganism derived labile enzymes, are stabilized by adding thereto a stabilizing agent comprising a protein fraction obtained by extracting yeast with water at 80.degree. to 100.degree. C., mixing the clear extract with 5 - 20% wt./vol. trichloracetic acid and separating the precipitate obtained.

Stabilizing agent for enzymes

Cholesterol oxidase which oxidizes cholesterol with oxygen to give cholestenone and hydrogen peroxide, is prepared by culturing Nocardia erthropolis ATCC 17895, Nocardia erthropolis ATCC 4277, Nocardia formica ATCC 14811 or Proactinomyce erythropolis NCIB 9158 first on a peptone-containing mineral salt medium and thereafter on at least one sterol compound as the sole source of carbon, and then isolating the cholesterol oxidase formed from the cultured micro-organism, wherein said sterol compound is of the formula ##STR1## in which R.sub.1 and R.sub.2 are hydrogen or together represent a double bond,

Process for the preparation of cholesterol oxidase

Carrier-bound proteins are prepared by reacting a protein in aqueous solution with a coupling compound having at least one group capable of acylating or alkylating proteins and at least one additional group capable of producing a bond with a carrier material, and reacting the resulting product with a carrier material, optionally forming the carrier material in situ by polymerization of a monomer or monomer mixture in the presence of said product.

Carrier-bound protein prepared by reacting the protein with an acylating

NADP-dependent dehydrogenases are separated and purified by subjecting mixtures containing it to affinity chromatography using as the affinity chromatography material a nucleotide with a phosphate group in the 2'- or 3'-position, bound to a solid carrier.

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Tutzing, Germany

Klaus Beaucamp