Boehringer Mannheim Gmbh

Gunter Lang

Wolfgang Gruber

Klaus Beaucamp

Peter Roeschlau

Erich Bernt

Hans Ulrich Bergmeyer

Joachim Ziegenhorn

Hans Seidel

Michael Jarsch

Albert Roder

Helmut Burtscher

Michael Nelboeck-Hochstetter

Friedrich Popp

Hans-Willi Krell

Ulfert Deneke

Knut Bartl

Helmgard Gauhl

Hans Mollering

Alexander Hagen

Klaus Kaluza

Michael Nelbock-Hochstetter

Gerhard Michal

Hans-Otto Beutler

Hellmuth Vetter

Detlef Von Hoerschelmann

Gotthilf Naher

Waldemar Thum

Ulrich Nagele

Ingo Bohn

Johanna Gramsall

Gunter Holz

Josef Danninger

A recombinant protein with D-hydantoinase activity which has the amino acid sequence SEQ ID NO 1 is obtainable in large amounts and has an improved temperature stability.

Recombinant D-hydantoinase, a process for the production and use

Enzyme specific for bilirubin which has a phenol oxidase activity of less than 0.5% and an activity for biliverdin of less than 10%, a broad pH optimum as well as a good thermostability. The enzyme is obtainable from plants such as alfalfa and is suitable for the determination as well as for the degradation of bilirubin in particular in biological liquids.

Bilirubin oxidase from alfalfa and use of the enzyme

The invention concerns a glucose-6-phosphate dehydrogenase which contains the amino acid sequence shown in SEQ ID NO:1 as well as a DNA coding for it and a process for the isolation of an enzyme according to the present invention.

Cloning and overexpression of glucose-6-phosphate dehydrogenase from

The invention concerns a new NAD.sup.+ -dependent D-malate dehydrogenase (decarboxylating, E.C. 1.1.1.83) from E. coli which consists of two subunits which each have a molecular weight of about 42 kD (SDS gel) and processes for the isolation of this enzyme from E. coli and for its stabilisation. The invention concerns in addition a method for the determination of D-malate with the D-malate dehydrogenase according to the present invention as well as a reagent for determining D-malate.

Enzymatic reagent for D-malate determination

For the determination of glycerol, the latter is incubated with galactose oxidase in an aqueous medium in the presence of oxygen and either the oxygen consumption or the amount of H.sub.2 O.sub.2 or glyceraldehyde that is formed is determined. A reagent suitable for this purpose consists of galactose oxidase and a system for the determination of H.sub.2 O.sub.2 or a system for the determination of glyceraldehyde. It can additionally contain an agent for the saponification of esterified glycerol.

Method and reagent for the determination of glycerol

For the determination of glycerin by oxidation with oxygen in the presence of glycerinoxidase and measurement of the oxygen consumption or of the H.sub.2 O.sub.2 formation, a glycerinoxidase from Aspergillus spec. DSM 1729 is used. A reagent suitable for this method consists of glycerinoxidase from Aspergillus spec. DSM 1729 and a system for the determination of H.sub.2 O.sub.2 and contains additionally, if desired, an agent for the saponification of esterified glycerin.

Method and reagent for the determination of glycerin

Storage-stable aqueous cholesterol oxidase compositions comprising cholestrol oxidase, a buffer and from 2 to 3.5 M sodium or potassium chloride as a stabilizing agent, are significantly superior to conventional compositions with respect to storage stability and fungibility.

Storage stable cholesterol oxidase compositions

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

In a process for the determination of total or bound cholesterol by liberating any bound cholesterol using cholesterol esterase and simultaneously or subsequently determining the liberated cholesterol and any initially free cholesterol with a cholesterol reacting enzyme, the improvement comprising using as the cholesterol reacting enzyme an NAD or NADP-dependent cholesterol dehydrogenase obtained from an anaerobic microorganism, e.g., Eubacterium sp. ATCC 21408, or from mammalian tissue, e.g., liver.

Composition and method for the determination of cholesterol

In a process for the determination of substrates or enzyme activities in a test batch, utilizing a conventional redox reaction as a measurement reaction, the improvement comprising carrying out the process in the presence of ascorbate oxidase; the addition of ascorbate oxidase substantially eliminates the interfering effect of ascorbic acid frequently found in the test sample.

Method for the determination of substrates or enzyme activities

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

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,

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Tutzing, Germany

Gunter Lang