Abstract: Described are compositions, in particular lyophilizates, containing proteolytic enzymes, and methods for producing the compositions. Typically these compositions contain one or more proteases with collagenase activity and a neutral protease, for example, thermolysis. The compositions are free of acetate salts. Surprisingly, such compositions can be dissolved in water more rapidly than lyophilized protease mixtures of the state of the art.

Abstract: The present invention provides improved variants of T7 RNA polymerase by introducing novel mutations which lead to improved thermostability of the enzyme. According to the invention, amino acid substitutions at the positions Val426, Ser633, Val650, Thr654, Ala702, Val795, and combinations thereof are advantageous.

Abstract: The present invention relates to a formulation of a thermostable DNA polymerase which is completely free of detergents and its particular use in real time polymerase chain reaction (PCR). Such a formulation may be obtained if the selected purification method does not require the addition of a detergent at any purification step.

Abstract: The present invention provides a method for purifying Clostridium histolyticum collagenase type I and type II proteins from a complex mixture by subsequently performing a precipitation with ammonium sulfate, hydrophobic interaction chromatography, cation exchange chromatography, and anion chromatography. Conditions are provided which lead to a stabilized, partially purified preparation even after the precipitation step. The method of the invention leads to a quick and efficient removal of other proteolytic activities. The preparations according to the invention provide exceptionally pure and intact collagenase type I and type II proteins which are enzymatically active. The invention also provides blends of the two isolated proteins. The invention further provides the use of the purified collagenase proteins or blends thereof for treating a tissue sample in vitro.

Abstract: The present invention provides a method for purifying Clostridium histolyticum collagenase type I and type II proteins from a complex mixture by subsequently performing a precipitation with ammonium sulfate, hydrophobic interaction chromatography, cation exchange chromatography, and anion chromatography. Conditions are provided which lead to a stabilized, partially purified preparation even after the precipitation step. The method of the invention leads to a quick and efficient removal of other proteolytic activities. The preparations according to the invention provide exceptionally pure and intact collagenase type I and type II proteins which are enzymatically active. The invention also provides blends of the two isolated proteins. The invention further provides the use of the purified collagenase proteins or blends thereof for treating a tissue sample in vitro.

Abstract: The present invention provides an aqueous composition comprising a protein with enzymatic activity of alpha-galactosidase. The present invention further provides a method of stabilizing an aqueous composition comprising a protein with enzymatic activity of alpha-galactosidase, and a method of preparing a purified aqueous composition comprising the protein with enzymatic activity of alpha-galactosidase.

Abstract: A method for producing solid proteinase K which is insoluble in water and has high purity, the method comprising the steps of adding to an aqueous solution of proteinase K ammonium sulfate in an amount of 0.1 to 0.2 M per 5 minutes up to a final amount of 1.5 to 2 M after 1 hour, thereby precipitating the proteinase K as a solid, and isolating the solid proteinase K.

Abstract: The present invention deals with the proteolytic enzyme thermolysin which tends to be unstable in aqueous solution. The invention provides methods and compositions to enhance the stability of dissolved thermolysin in aqueous solution. Thermolysin, crude thermolysin or a lyophilisate containing thermolysin and one or more salts, is contacted with an aqueous buffer with a low salt concentration and a first solution is formed. Subsequently, a further salt in solid form is added and dissociated, thereby forming a second solution comprising thermolysin in a stabilized form.

Abstract: The present invention provides improved media for the cultivation of Clostridium histolyticum and culture supernatants for the biotechnological production of collagenase enzymes. The nutrient media according to the invention comprise one or more peptones from a non-mammalian source, preferably plant-derived peptones. The media can additionally comprise fish gelatin. The invention provides media, culture supernatants comprising Clostridium histolyticum collagenase, and methods to produce said collagenase.

Abstract: The present invention concerns a method for producing recombinant trypsin from porcine pancreas in Pichia pastoris which is soluble and secreted into the culture medium, whereby expression at pH 3.0-4.0 substantially prevents activation of trypsinogen to ?-trypsin and autolysis of ?-trypsin by ?-trypsin into ?-trypsin and from there into inactive peptides.

Abstract: The invention concerns a method for the expression of a gene coding for a soluble proteinase K in yeast e.g. in Pichia pastoris with subsequent secretion into the culture medium. In addition a method for purifying the heterologously expressed and secreted proteinase K is described.

Abstract: The present invention concerns a method for producing recombinant trypsin from porcine pancreas in Pichia pastoris which is soluble and secreted into the culture medium, whereby expression at pH 3.0-4.0 substantially prevents activation of trypsinogen to ?-trypsin and autolysis of ?-trypsin by ?-trypsin into ?-trypsin and from there into inactive peptides.

Abstract: The invention concerns a method for the recombinant production or expression of eukaryotic alkaline phosphatase mutants in yeast cells wherein the specifically introduced mutations result in a reduction of the specific AP activity by at least a factor 1:100. The invention also concerns a method for inserting corresponding nucleic acid sequences into a vector for expression in methylotrophic yeast strains and it concerns corresponding vectors and host strains.

Abstract: The present invention concerns a method for producing recombinant trypsin from porcine pancreas in Pichia pastoris which is soluble and secreted into the culture medium, whereby expression at pH 3.0-4.0 substantially prevents activation of trypsinogen to ?-trypsin and autolysis of ?-trypsin by ?-trypsin into ?-trypsin and from there into inactive peptides.

Abstract: A method for producing solid proteinase K which is insoluble in water and has high purity, the method comprising the steps of adding to an aqueous solution of proteinase K ammonium sulfate in an amount of 0.1 to 0.2 M per 5 minutes up to a final amount of 1.5 to 2 M after 1 hour, thereby precipitating the proteinase K as a solid, and isolating the solid proteinase K.

Abstract: The invention concerns a method for the recombinant production or expression of eukaryotic alkaline phosphatase mutants in yeast cells wherein the specifically introduced mutations result in a reduction of the specific AP activity by at least a factor 1:100. The invention also concerns a method for inserting corresponding nucleic acid sequences into a vector for expression in methylotrophic yeast strains and it concerns corresponding vectors and host strains.

Abstract: The invention concerns recombinant proteinase K. Furthermore a method for producing recombinant proteinase K is disclosed, which is characterized in that a) a host cell is transformed with a recombinant nucleic acid which codes for the zymogenic precursor of proteinase K, b) the host cell is cultured in such a manner that the zymogenic precursor of proteinase K is formed in the form of inclusion bodies in the host cell, c) the inclusion bodies are isolated and natured under conditions which result in the formation of the protease part of the zymogenic precursor in its natural conformation, d) the natured proteinase K is activated and purified.

Abstract: The invention concerns a method for the recombinant production or expression of eukaryotic alkaline phosphatase mutants in yeast cells wherein the specifically introduced mutations result in a reduction of the specific AP activity by at least a factor 1:100. The invention also concerns a method for inserting corresponding nucleic acid sequences into a vector for expression in methylotrophic yeast strains and it concerns corresponding vectors and host strains.

Abstract: A method is provided for the production of a bovine pancreatic protein with a specific desoxyribonuclease activity of at least 6,000 units per mg of protein. Methylotrophic yeast is used as a heterologous host organism. The bovine pancreatic protein is secreted into the growth medium from which it is purified.

Abstract: The invention provides a method to produce a protein with carboxypeptidase B activity from a pro-carboxypeptidase B zymogen, derived from a non-animal host organism. Carboxypeptidase B is activated from the zymogen using non-denaturing conditions. Particularly, the activation is performed under conditions that avoid unwanted non-covalent binding of the propeptide to the activated carboxypeptidase B enzyme.