Boehringer Mannheim Gmbh

Hoffmann-la Roche Inc.

Roche Diagnostics Gmbh

Dorothee Ambrosius

Rainer Rudolph

Friederike Hesse

Helmut Burtscher

Elisabeth Schwarz

Joerg Schaeffner

Erhard Kopetzki

Peter Sluka

Hans-Joachim Guder

Klaus Kaluza

Bernd Schmidt

Petra Ruger

Stephan Gross

A polypeptide with the properties of collagenase class I from Clostridium histolyticum (CHC I) and the amino acid sequence SEQ ID NO:2 which is optionally N-terminally extended by one or several amino acids of the sequence SEQ ID NO:3 is advantageously suitable for the isolation of cells from mammalian tissue and human tissue.

Recombinant collagenase type I from clostridium histolyticum and its use for isolating cells and groups of cells

A process for the production of a naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges by a) culturing prokaryotic cells in which the said prokaryotic cells contain an expression vector which codes for the said polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) secreting the polypeptide into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium, which is characterized in that a nucleic acid coding for a molecular chaperone is additionally expressed in the said prokaryotic cell and the chaperone is secreted into the periplasm, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

Process for the production of naturally folded and secreted proteins by co-secretion of molecular chaperones

A process produces a water-soluble, naturally folded eukaryotic polypeptide containing two or several cysteines linked by disulfide bridges. This process involves culturing prokaryotic cells, a) in which the prokaryotic cells contain an expression vector which encodes the polypeptide which contains a prokaryotic signal sequence at the N-terminus, b) under conditions under which the polypeptide is secreted into the periplasm or the medium, c) cleaving the signal sequence and isolating the polypeptide from the periplasm or the medium. In this process, the culturing is carried out in the presence of arginine or a compound of the formula I R,&mdash;CO&mdash;NR,(I) in which R and R,represent hydrogen or a saturated or unsaturated branched or unbranched C,-C,alkyl chain and R,represents hydrogen, NHR,or a saturated or unsaturated branched or unbranched C,-C,alkyl chain, is suitable for the recombinant production of polypeptides in prokaryotes in a high yield.

Process for the production of naturally folded and secreted proteins

Oxygen-independent cholesterol-converting enzyme obtainable from Actinomycetes or Basidiomycetes which utilizes artificial electron acceptors as well as a method and reagent for the determination of (total) cholesterol using the enzyme.

Microbial cholesterol dehydrogenase, process for its production and use

Recombinant proteinase from clostridium histolyticum and its use for

An electrochemical sensor is provided containing a supporting material having a surface of noble metal such as gold or palladium to which is bound an enzyme such as glucose oxidase via binding molecules. The binding molecules are in the form of a homogenous monolayer and are bound to the metal surface by anchor groups such as thiol, disulphide or phosphine groups. The enzyme is linked to the binding molecules by an ionic bond, a covalent bond or a metal-chelate bond. The binding molecules may contain a spacer such as an alkylene chain. Coverage of the binding molecules on the metal surface is less than the maximum coverage to optimize enzyme coverage, decrease response time of the sensor and increase signal yield per surface area. The monolayer may contain diluent molecules which are not linked to an enzyme and which are bound to the metal surface via anchor groups. The diluent molecules lower the concentration of binding molecules in the monolayer, increase conductivity of the monolayer and lower unspecific binding of components from a sample solution in contact with the sensor. The binding molecules can be linked by an ionic bond or a metal-chelate bond, respectively, to a charged amino acid residue or a chelate-forming amino acid residue of the N-terminus, C-terminus, or N- and C-terminus of recombinant enzyme protein.

Electrochemical sensor containing an enzyme linked to binding molecules

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Munich, Germany

Dorothee Ambrosius