Abstract: A novel enzyme which has an activity to release side chain carboxyl groups and ammonia from a protein by acting upon side chain amido groups in the protein. This invention relates to a method for the production of an enzyme, which comprises culturing in a medium a strain that belongs to a bacterium classified into Cytophagales or Actinomycetes and has the ability to produce an enzyme having a property to deamidate amido groups in protein, thereby effecting production of said enzyme, and subsequently collecting said enzyme from the culture mixture. It also relates to a method for the modification of protein making use of a novel enzyme which directly acts upon amido groups in protein as well as to an enzyme which has a property to deamidate amido groups in protein and a gene which encodes said enzyme.

Abstract: The object of this invention is to provide a new cysteine protease. The object is achieved by providing a new cysteine protease which is obtained from a flesh fly (Sarcophaga peregrina) and comprises 26 kDa and 29 kDa subunits.

Abstract: Bacillus licheniformis O.W.U. 138B and 88B and other bacterial strains capable of degrading feathers isolated from wild birds, are disclosed. The strains and the keratinases produced by these strains are useful for degrading waste feathers produced by commercial poultry processing; producing animal feed, fertilizer, or natural gas from poultry waste; and cleaning of certain fabrics.

Abstract: A method for preparing a crystalline protease is provided which comprises preparing an aqueous solution containing the protease enzyme and adding to the aqueous solution sodium sulfate, allowing the crystallization to take place at a temperature between 10° C. and 60° C.

Abstract: Bioprocesses are disclosed for the production of compounds which can be produced from a dipeptide intermediate. The process comprises production of a recombinant polypeptide which contains the dipeptide intermediate. The dipeptide intermediate is further processed to ultimately provide the finished product.

Abstract: A psychrophilic protease is here disclosed which has the following physicochemical properties: (a) specific activity and substrate specificity: the protease acts on casein, gelatin, hemoglobin and albumin to specifically decompose them, and the substrate specificity decreases in the order of casein, gelatin, hemoglobin and albumin; (b) optimal pH: 8.0; (c) pH stability: the protease is stable in the range of pH 6.5 to pH 10.0 at 30° C. for 1 hour; (d) optimal temperature: about 40° C.; (e) temperature stability: at pH 7 for 1 hour, the protease is active at a temperature up to 30° C., but it is inactivated at 40° C. as much as about 40% and completely inactivated at 50° C. in about 10 minutes; (f) enzyme activity: the protease has 50% or more of its maximum activity at about 20° C.; (g) the active center of the enzyme is serine; and (h) the molecular weight of the protease is about 70 kDa as measured by SDS-PAGE.

Abstract: The present invention describes a novel polypeptide, and methods of its use in effective thrombolytic therapy in the treatment of coronary and pulmonary thrombosis. Its use is also disclosed in vaccines to abrogate a streptococcal infection. Pharmaceutical compositions containing the novel polypeptide are included. One particular form of the novel polypeptide is streptococcal surface enolase (SEN), a specific binding protein for human plasmin and/or human plasminogen on group A streptococci that displays classical &agr;-enolase activity, i.e., it can catalyze the dehydration of D-glycerate-2-phosphate to phosphoenolpyruvate. In addition, SEN impedes the inhibition of the fibrinolytic activity of plasmin by &agr;2-antiplasmin and can bind plasminogen without preventing streptokinase from cleaving this plasmin precursor.

Abstract: A purified xylanase derived from B. Pumilus PRL B12 is disclosed. This xylanase is efficient for use in the biobleaching of wood pulp, permitting a strong reduction in the quantity of chlorine used and AOX compounds produced in classical and ECF wood pulp bleaching sequences as well as the quantity of ozone used in TCF sequences. The gene coding for the xylanase was isolated and purified and used to construct an expression vector therefor. A recombinant host strain of B. licheniformis is also disclosed which is efficient for expressing heterologous enzymes, including the xylanase when transformed by the expression vector.

Abstract: The invention relates to a method of producing an extracellular protein in a bacterium provided with an inner and an outer cell membrane, the method comprising: (a) providing a recombinant vector including a DNA construct comprising a DNA sequence encoding the prepropeptide or part of the prepropeptide of a bacterial extracellular protease selected from the group consisting of Achromobacter lyticus protease I, Bacillus metalloproteases and Bacillus serine proteases preceding and operably connected to a DNA sequence encoding a desired protein, (b) transforming cells of a microorganism provided with an inner and outer cell membrane with the recombinant vector of step (a), (c) culturing the transformed cells of step (b) under conditions permitting expression of said DNA insert and leakage of the bacterial extracellular protease propeptide fused to the desired protein into the culture medium, and (d) recovering the resulting protein from the medium.

Abstract: Disclosed is an alkaline protease having a specific activity of 214,000 (U/mg protein) when using casein as a substrate and 52,700 (U/mg protein) when using keratin as a substrate. The alkaline protease is produced by a strain belonging to alkalophilic actinomycetes, Streptomyces, particluraly Streptomyces sp. TOTO-9305 strain (FERM P-13640).

Abstract: An isolated and purified analog of Haemophilus influenzae Hin47 protein has a decreased protease activity which is less than about 10% of that of natural Hin47 protein and preferably substantially the same immunogenic properties as natural Hin47 protein. An isolated an purified nucleic acid molecule encoding the Hin47 analog may be provided in a recombinant plasmid which may be introduced into a cell which is grown to produce the Hin47 analog. Immunogenic compositions comprising the Hin47 analog and the encoding nucleic acid may be formulated as vaccines for in vivo administration to a host, including a human, to confer protection against diseases caused by a bacterial pathogen, including Haemophilus species, such as Haemophilus influenzae, that produces Hin47 protein or a protein capable of inducing antibodies in the host specifically reactive with Hin47 protein. The Hin47 analog and the encoding nucleic acid also may be employed in diagnostic applications.

Abstract: The subject invention pertains to new thermostable enzymes and the use of these enzymes both in proteolysis as well as protein and polypeptide synthesis. The subject invention further concerns polynucleotide sequences which encode the enzymes of the subject invention.

Abstract: The invention provides isolated nucleic acid compounds encoding FtsH of Streptococcus pneumoniae. Also provided are vectors and transformed host cells for expressing the encoded protein, and a method for identifying compounds that bind and/or inhibit said protein.

Abstract: Mutant B. lentus DSM 5483 proteases are derived by the replacement of at least one amino acid residue of the mature form of the B. lentus DSM 5483 alkaline protease. The mutant proteases are expressed by genes which are mutated by site-specific mutagenesis. The amino acid sites selected for replacement are identified by means of a computer based method which compares the three dimensional structure of the wild-type protease and a reference protease.

Abstract: An acyl peptide hydrolase having an optimum temperature range of 90-95.degree. C. and a gene encoding the same are disclosed. With the above enzyme, it becomes possible to conduct amino terminal analysis of acylated proteins and peptides at high temperatures.

Abstract: The present invention relates to a process for producing optically active 3-quinuclidinol or derivatives, wherein a racemic 3-quinuclidinol ester represented by the general formula (I): ##STR1## wherein R represents a straight-chain or branched alkyl group, and (H.sup.+) represents that said ester may be in the form of a salt formed with a mineral acid or an organic acid, is reacted with a microorganism belonging to the genus Aspergillus, Rhizopus, Candida or Pseudomonas having the ability to asymmetrically hydrolyze said ester linkage, a culture of said microorganism, a treated material from said microorganism, an enzyme produced by said microorganism, or an enzyme derived from swine or cattle.According to the present invention, there is provided a process for easily producing optically active 3-quinuclidinol derivatives which are important synthetic intermediates for pharmaceutical preparations etc.

Abstract: The invention relates to new regenerable filtration adjuvants which are usable for the filtration of liquids, particularly beer at the end of the secondary fermentation storage, characterized in that they comprise synthetic or natural incompressible polymer grains or natural incompressible grains grains having a sphericity coefficient varying between approximately 0.6 and 0.9. The invention also relates to a method for regenerating a filtration adjuvant.

Abstract: The object of this invention is to provide a new cysteine protease. The object is achieved by providing a new cysteine protease which is obtained from a flesh fly (Sarcophaga peregrina) and comprises 26 kDa and 29 kDa subunits.

Abstract: The present invention relates to a method for crystallization of a protein obtained from a protein-containing solution which involves (a) treating the protein-containing solution with a salt containing a sulphur atom having an oxidation state less than 6, and (b) recovering the protein in crystalline form.

Abstract: A process for the production of a desired polypeptide comprising the steps of: (1) transforming host cells with an expression vector comprising a gene coding for a fusion protein comprising a desired polypeptide and a protective polypeptide; (2) culturing the transformed host cells so as to express said gene to produce a fusion protein; and (3) excising the desired polypeptide from the fusion protein with a protease intrinsic to the host cells. According to the present invention, a large amount of a desired polypeptide can be produced at a low cost. Especially according to the present invention, a large amount of S. aureus V8 protease can be efficiently produced at low cost using a safe host such as E. coli according to gene recombination procedures.

Abstract: An isolated and purified analog of Haemophilus influenza Hin47 protein has a decreased protease activity which is less than about 10% of that of natural Hin47 protein and preferably substantially the same immunogenic properties as natural Hin47 protein. An isolated and purified nucleic acid molecule encoding the Hin47 analog may be provided in a recombinant plasmid which may be introduced into a cell which is grown to produce the Hin47 analog. Immunogenic compositions comprising the Hin47 analog and the encoding nucleic acid may be formulated as vaccines for in vivo administration to a host, including a human, to confer protection against diseases caused by a bacterial pathogen, including Haemophilus species, such as Haemophilus influenzae, that produces Hin47 protein or a protein capable of inducing antibodies in the host specifically reactive with Hin47 protein. The Hin47 analog and the encoding nucleic acid also may be employed in diagnostic applications.

Abstract: An isolated and purified analog of Haemophilus influenzae Hin47 protein has a decreased protease activity which is less than about 10% of that of natural Hin47 protein and preferably substantially the same immunogenic properties as natural Hin47 protein. An isolated an purified nucleic acid molecule encoding the Hin47 analog may be provided in a recombinant plasmid which may be introduced into a cell which is grown to produce the Hin47 analog. Immunogenic compositions comprising the Hin47 analog and the encoding nucleic acid may be formulated as vaccines for in vivo administration to a host, including a human, to confer protection against diseases caused by a bacterial pathogen, including Haemophilus species, such as Haemophilus influenzae, that produces Hin47 protein or a protein capable of inducing antibodies in the host specifically reactive with Hin47 protein. The Hin47 analog and the encoding nucleic acid also may be employed in diagnostic applications.

Abstract: Hydrophillic compositions and methods of use are provided for debriding and wound healing applications. The compositions contain certain proteases produced by microorganisms of the genus Vibrio.

Abstract: Provided herein is a Porphyromonas gingivalis high molecular weight arginine-specific proteinase comprising a protease component of 50 kD and a hemagglutinin component of about 44 kD as estimated by SDS-PAGE. The proteinase is stimulated by glycine containing peptides and glycine analogues. It is inhibited by cysteine protease group specific inhibitors.

Abstract: A purified enzyme mixture useful for isolating cells or cell clusters from tissue is disclosed. The mixture includes at least two collagenase enzymes, at least two other proteases, and additional non-protease components. The mixture is purified by removing at least some of the non-protease components. The purified mixture may then be used to isolate cells or cell clusters from tissue. Also disclosed are the essential components of the purified enzyme mixture as well as preferred ranges and ratios of these essential components for isolating cells or cell clusters from tissue. Finally, a dissociation system is disclosed that can be used with the purified enzyme mixture to dissociate tissue and recover cells or cell clusters.

Abstract: The invention relates to an agent specific for peripheral sensory afferents. The agent may inhibit the transmission of signals between a primary sensory afferent and a projection neuron by controlling the release of at least one neurotransmitter or neuromodulator from the primary sensory afferent. The agent may be used in or as a pharmaceutical for the treatment of pain, particularly chronic pain.

Abstract: The present invention relates to isolated nucleic acid sequences encoding polypeptides having tripeptide aminopeptidase activity. The invention also relates; to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as recombinant methods for producing the polypeptides.

Abstract: Mutant B. lentus DSM 5483 proteases are derived by the replacement of at least one amino acid residue of the mature form of the B. lentus DSM 5483 alkaline protease. The mutant proteases are expressed by genes which are mutated by site-specific mutagenesis. The amino acid sites selected for replacement are identified by means of a computer based method which compares the three dimensional structure of the wild-type protease and a reference protease.

Abstract: Dihydroorotate dehydrogenase (DHOD) inhibititon assays are utilized in a method for identifying new fungicides; DHOD inhibitors are useful in fungicidal methods and compositions. Resistance genes that are mutants of the wild-type Aspergillus nidulans dihydroorotate dehydrogenase gene have been isolated and found to impart resistance to certain DHOD inhibitors. The resistance genes are useful as selectable markers in fungi. The wild-type gene is also provided.

Abstract: spsB polypeptides and DNA (RNA) encoding such spsB and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such spsB for the treatment of infection, particularly bacterial infections. Antagonists against such spsB and their use as a therapeutic to treat infections, particularly bacterial infections are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to the presence of spsB nucleic acid sequences and the polypeptides in a host. Also disclosed are diagnostic assays for detecting polynucleotides encoding spsB and for detecting the polypeptide in a host.

Abstract: The invention relates to an isolated nucleic acid encoding Streptococcus pneumoniae IgA protease and an isolated polypeptide comprising Streptococcus pneumoniae IgA protease and methods of use thereof.

Abstract: Novel carbonyl hydrolase mutants derived from the amino acid sequence of naturally-occurring or recombinant non-human carbonyl hydrolases and DNA sequences encoding the same. The mutant carbonyl hydrolases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant carbonyl hydrolase to encode the substitution, insertion or deletion of one or more amino acids in the amino acid sequence of a precursor carbonyl hydrolase. Such mutants have one or more properties which are different than the same property of the precursor hydrolase.

Abstract: The invention provides a group of subtilisin YaB mutants, which are obtained by the substitution of the glycine residues at the 124, 151 and 159 positions of wild type subtilisin YaB with other amino acid residues by site directed mutagenesis. The subtilisin YaB mutants have special substrate specificity and relatively higher elastin/casein hydrolyzing activity. The subtilisin YaB mutants can be used in various aspects such as the quality improvement of meat and the protein processing for foodstuff and feedstuff. Moreover, the invention also includes the nucleic acid sequences encoding such enzyme mutants and the uses of such enzyme mutants.

Abstract: The invention relates to an enzyme which cleaves surface proteins of gram-positive bacteria, to methods of detecting the enzyme, and methods of isolating the enzyme. In particular, the enzyme is isolated from a group A Streptococcus, and cleaves at the sequence LPXTGX (SEQ ID NO:1). A method for screening putative inhibitors of the enzyme which cleaves the anchor region of surface proteins from gram positive bacteria is also disclosed.

Abstract: The present invention concerns a process for the isolation of recombinant IgA protease from inclusion bodies. In addition a recombinant DNA is claimed which codes for an IgA protease whose C-terminal helper sequence and preferably also its N-terminal signal sequence is no longer active.

Abstract: The invention provides licB polypeptides and polynucleotides encoding licB polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing licB polypeptides to screen for antibacterial compounds.

Abstract: Novel carbonyl hydrolase mutants derived from the amino acid sequence of naturally-occurring or recombinant non-human carbonyl hydrolases and DNA sequences encoding the same. The mutant carbonyl hydrolases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant carbonyl hydrolase to encode the substitution, insertion or deletion of one or more amino acids in the amino acid sequence of a precursor carbonyl hydrolase. Such mutants have one or more properties which are different than the same property of the precursor hydrolase.

Abstract: Methods for the identification, production and use of derivatives of the invention obtained by preparing a DNA fragment comprising at least the part of the coding sequence of staphylokinase that provides for its biological activity; performing in vitro site-directed mutagenesis on the DNA fragment to replace one or more codons for wild-type amino acids by a codon for another amino acid; cloning the mutated DNA fragment in a suitable vector; transforming or transfecting a suitable host cell with the vector; and culturing the host cell under conditions suitable for expressing the DNA fragment. Preferably the DNA fragment is a 453 bp EcoRI-HindIII fragment of the plasmid pMEX602sakB, the in vitro site-directed mutagenesis is performed by spliced overlap extension polymerase chain reaction and the mutated DNA fragment is expressed in E. coli strain TG1 or WK6.

Abstract: An enzyme composition and its use for isolating cells or cell clusters from tissue is disclosed. The composition includes two purified collagenase enzymes and an endoprotease. The ratio of the total FITC casein activity of the enzyme composition to the Wunsch units of activity of the masses of collagenase I and collagenase II is about 85:1 to about 3,900:1.

Abstract: The present invention relates to novel proteolytic enzymes. More specifically, the present invention relates to proteolytic enzymes obtainable from strains of Amycolata and Amycolatopsis. Moreover the invention relates to a process for the preparation of the proteolytic enzyme of the invention, as well as detergent additives and detergent compositions comprising the proteolytic enzyme.

Abstract: An isolated and purified analog of Haemophilus influenzae Hin47 protein has a decreased protease activity which is less than about 10% of that of natural Hin47 protein and preferably substantially the same immunogenic properties as natural Hin47 protein. An isolated an purified nucleic acid molecule encoding the Hin47 analog may be provided in a recombinant plasmid which may be introduced into a cell which is grown to produce the Hin47 analog. Immunogenic compositions comprising the Hin47 analog and the encoding nucleic acid may be formulated as vaccines for in vivo administration to a host, including a human, to confer protection against diseases caused by a bacterial pathogen, including Haemophilus species, such as Haemophilus influenzae, that produces Hin47 protein or a protein capable of inducing antibodies in the host specifically reactive with Hin47 protein. The Hin47 analog and the encoding nucleic acid also may be employed in diagnostic applications.

Abstract: A hybrid botulinal neurotoxin is disclosed. In one embodiment, the neurotoxin comprises a combination of a botulinal neurotoxin heavy chain and light chain, wherein the light chain and heavy chain are not of the same serotype. A method for creating hybrid neurotoxins comprised of different functional domains is also disclosed.

Abstract: An isolated and purified analog of Haemophilus influenzae Hin47 protein has a decreased protease activity which is less than about 10% of that of natural Hin47 protein and preferably substantially the same immunogenic properties as natural Hin47 protein. An isolated an purified nucleic acid molecule encoding the Hin47 analog may be provided in a recombinant plasmid which may be introduced into a cell which is grown to produce the Hin47 analog. Immunogenic compositions comprising the Hin47 analog and the encoding nucleic acid may be formulated as vaccines for in vivo administration to a host, including a human, to confer protection against diseases caused by a bacterial pathogen, including Haemophilus species, such as Haemophilus influenzae, that produces Hin47 protein or a protein capable of inducing antibodies in the host specifically reactive with Hin47 protein. The Hin47 analog and the encoding nucleic acid also may be employed in diagnostic applications.

Abstract: The present invention relates to isolated nucleic acid sequences encoding polypeptides having protease activity, in which the polypeptides are obtainable from an alkalophilic Bacillus species having enhanced stability towards bleaching agents of the peroxy type. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as recombinant methods for producing the polypeptides.

Abstract: The present invention relates to methods of producing a polypeptide, comprising: (a) introducing into a respiratory-defective mutant of a cell (i) one or more first nucleic acid sequences which complement the respiratory defect and (ii) a second nucleic acid sequence which encodes the polypeptide; (b) cultivating the cell containing the first and second nucleic acid sequences in a culture medium under aerobic conditions suitable for expression of the first and second nucleic acid sequences; and (c) isolating the polypeptide from the cultivation medium of the cell. The present invention also relates to methods for disrupting a gene in a respiratory-deficient mutant cell. The present invention further relates to respiratory-deficient mutant cells and methods for obtaining such mutant cells.

Abstract: lep polypeptides and DNA (RNA) encoding such lep and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such lep for the treatment of infection, particularly bacterial infections. Antagonists against such lep and their use as a therapeutic to treat infections, particularly bacterial infections are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to the presence of lep nucleic acid sequences and the polypeptides in a host. Also disclosed are diagnostic assays for detecting polynucleotides encoding leader peptidase and for detecting the polypeptide in a host.

Abstract: D1 protease has been isolated from the alga (Scenedesmjus obliquus), wheat, and Synechocystis PCC 6803 and the genes encoding these enzymes have been cloned and sequenced. Native or recombinantly produced enzyme has been used to develop assays to detect herbicidal compositions capable of inhibiting the D1 protease enzyme.

Abstract: Bacillus licheniformis O.W.U. 138B and 88B and other bacterial strains capable of degrading feathers isolated from wild birds, are disclosed. The strains and the keratinases produced by these strains are useful for degrading waste feathers produced by commercial poultry processing; producing animal feed, fertilizer, or natural gas from poultry waste; and cleaning of certain fabrics.

Abstract: An isolated and purified analog of Haemophilus influenzae Hin47 protein has a decreased protease activity which is less than about 10% of that of natural Hin47 protein and preferably substantially the same immunogenic properties as natural Hin47 protein. An isolated an purified nucleic acid molecule encoding the Hin47 analog may be provided in a recombinant plasmid which may be introduced into a cell which is grown to produce the Hin47 analog. Immunogenic compositions comprising the Hin47 analog and the encoding nucleic acid may be formulated as vaccines for in vivo administration to a host, including a human, to confer protection against diseases caused by a bacterial pathogen, including Haemophilus species, such as Haemophilus influenzae, that produces Hin47 protein or a protein capable of inducing antibodies in the host specifically reactive with Hin47 protein. The Hin47 analog and the encoding nucleic acid also may be employed in diagnostic applications.

Abstract: The invention discloses PEG-modified proteases, compositions containing these modified enzymes, and methods for using them to clean contact lenses. The methods of the present invention are also directed to the simultaneous cleaning and disinfecting of contact lenses, when compositions of the present invention are combined with a suitable disinfectant.