Abstract: Methods for the identification, production and use of staphylokinase derivatives characterized by a reduced immunogenicity after administration in patients. The derivatives of the invention are 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; culturing the host cell under conditions suitable for expressing the DNA fragment; and purifying the expressed staphylokinase derivative to homogeneity. Preferably the DNA fragment is a 453 bp EcoRI-HindIII fragment of the plasmid pMEX602sakB, (pMEX.

Abstract: A portion of the lysostaphin gene of Staphylococcus simulans has been cloned and overexpressed in the cytoplasm of E. coli to yield lysostaphin, in the absence of preprolysostaphin and prolysostaphin, under the transcriptional control of an IPTG-inducible promoter and a ribosome binding site. IPTG induction of the transformed host cells produces intracellular, soluble, mature lysostaphin (27 kDa), in the complete absence of preprolysostaphin and prolysostaphin. The mature lysostaphin so formed dose not require post-translational modification. The mature lysostaphin so formed can be used treat and prevent staphylococcal infections.

Abstract: The present invention relates to a novel methods and compositions for producing hyper and hypo allergenic compositions. Specifically, the present invention comprises neutralizing or reducing the ability of T-cells to recognize epitopes and thus prevent sensitization of an individual to the protein. Alternatively, T-cell epitopes are mutated to produce increased immunogenic reactions.

Abstract: The present invention relates to subtilase variants having a reduced tendency towards inhibition by substances present in eggs, such as trypsin inhibitor type IV-0. In particular, the variants comprise at least one additional amino acid residue between positions 42-43, 51-56, 155-161, 187-190, 216-217, 217-218 or 218-219 (in BASBPN numbering). These subtilase variants are useful exhibiting excellent or improved wash performance on egg stains when used in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dishwash composition, including automatic dishwash compositions. Also, isolated DNA sequences encoding the variants, expression vectors, host cells, and methods for producing and using the variants of the invention. Further, cleaning and detergent compositions comprising the variants are disclosed.

Abstract: Compounds and methods for designing and identifying compounds which inhibit TFPP-like aspartyl protease enzymes by targeting the aspartic acid residues of the active site or mimicking peptides corresponding to the region surrounding the substrate's cleavage site are provided. Agents identified as inhibitors of TFPP-like aspartyl proteases such as type 4 prepilin peptidases are expected to be useful as anti-bacterial agents and in inhibiting development of drug resistant strains of bacteria.

Abstract: The invention provides an enzymatic unhairing agent for use in an unhairing step in tanning for producing leather comprising an alkaline protease as an active component; a treatment solution comprising a pH-adjusting agent and the enzymatic unhairing agent; a method for enzymatic unhairing treatment in tanning for producing leather comprising contacting the treatment solution with a raw hide or skin; and a leather thus produced. According to the invention, it is attained markedly reduction of the pollution load in the unhairing waste water and leather and recovered hairs both of good quality can be obtained.

Abstract: The invention provides methods for identifying a modulator of quorum sensing signaling in bacteria, and for identifying a quorum sensing controlled gene in bacteria. In addition, the invention provides quorum sensing controlled genetic loci in Pseudomas aeruginosa. Novel indicator strains and vectors for engineering the strains for use in the method of the invention are also provided.

Abstract: There are provided hyperthermostable proteases having an amino acid sequences represented by SEQ ID NOs: 1, 3 and 5 of the Sequence Listing or functional equivalents thereof and hyperthermostable protease genes encoding those hyperthermostable protease. There is also disclosed a process for preparation of a hyperthermostable protease by culturing a transformant containing the gene.

Abstract: The present invention relates to a novel strain of alkalothermophilic Bacillus sp. isolated from a hot spring at Vajeshwari, District Thane, The State of Maharashtra, India and deposited at American Type Culture Centre (ATCC), bearing accession No. PTA 972, said strain of Bacillus sp. having the following characteristics (i) aerobic, (ii) gram positive, (iii) motile, (iv) spore forming, (v) capable of growing in a alkaline medium at pH 8-10, and (vi) exhibiting negative reaction towards production of indole, hydrogen, sulfide, ammonia and urease and positive reaction for hydrolysis of starch, production of catalase, hydrolysis of casein and reduction of nitrate.

Abstract: The present invention provides a process for economically producing N-acetylneuraminic acid without using expensive materials such as pyruvic acid and phosphoenolpyruvic acid. The process comprises: allowing (i) a culture of a microorganism having N-acetylneuraminic acid aldolase activity or N-acetylneuraminic acid synthetase activity, or a treated matter of the culture, (ii) a culture of a microorganism capable of producing pyruvic acid or a treated matter of the culture, or a culture of a microorganism capable of producing phosphoenolpyruvic acid or a treated matter of the culture, (iii) N-acetylmannosamine, and (iv) an energy source which is necessary for the formation of pyruvic acid or phosphoenolpyruvic acid to be present in an aqueous medium to form and accumulate N-acetylneuraminic acid in the aqueous medium; and recovering N-acetylneuraminic acid from the aqueous medium.

Abstract: An isolated oral bacterial polypeptide having amidolytic activity for cleavage of denatured polypeptides and nondenatured serpin polypeptides and particularly a human &agr;1-proteinase inhibitor polypeptide is provided. The mature polypeptide of the invention has a molecular weight of about 70 kD to about 80 kD. Also provided is an isolated nucleic acid sequence encoding the oral bacterial polypeptide of the invention, methods for identifying inhibitors of the polypeptide and compositions such as immunogenic compositions and inhibitor compositions.

Abstract: Vaccines, antibodies, proteins, DNAs and RNAs for diagnosis, prophylaxis, treatment and detection of Cryptosporidium species or Cryptosporidium species infections. Cryptosporidium species antigen and DNAs and RNA encoding the Cryptosporidium antigen and fragments thereof and recombinant proteins or fusion proteins produced thereby. Methods for diagnosis, prophylaxis, treatment and detection of Cryptosporidium species infections.

Abstract: The present invention relates to a novel enzyme (&agr;-GARE) which releases an amino acid residue having a glycated &agr;-amino group (&agr;-GA) from a glycated protein etc. and to bacterial strains producing the same. Examples of the bacterial strains include Sphingomonas parapaucimobilis KDK1004 (FERM BP-7041). The &agr;-GARE is contained in the culture supernatant of this strain and &agr;-GA can be released from a glycated peptide by using the same, as shown in FIG. 1.

Abstract: The invention concerns methods for preparing pristinamycin IIA or IIB.

Abstract: Novel protease variants derived from the DNA sequences of naturally-occurring or recombinant non-human proteases are disclosed. The variant proteases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant protease to generate the substitution of a plurality of amino acid residues in the amino acid sequence of a precursor protease. Such variant proteases have properties which are different from those of the precursor protease, such as altered wash performance.

Abstract: The present invention relates to mutations of a subtilisin gene which result in changes in the chemical characteristics of subtilisin enzymes. Mutations at specific nucleic acids of the subtilisin gene result in amino acid substitutions and consequently, altered enzyme function. Some of these mutant enzymes exhibit physical properties advantageous to industrial applications, particularly in the detergent industry, providing subtilisin which is more stable to oxidation, possesses greater protease activity, and exhibits improved washability.

Abstract: Novel proteins and their corresponding nucleotide sequences in enteroaggregative Escherichia coli (EAEC) are provided. In particular, Aap and the five gene cluster (aat) of the AA probe region of the pAA plasmid of EAEC 042 have been identified, sequenced, and further characterized. The use of these novel proteins and their corresponding nucleotide sequences for diagnosis, therapy, and prevention of EAEC infections is also provided.

Abstract: A new fibrinogen binding protein or polypeptide originating from coagulase negative staphylococci, biotechnological methods for producing the protein or polypeptide having fibrinogen binding activity and a recombinant DNA molecule coding for the protein (or fragments thereof), and micro-organisms (including viruses) containing this recombinant DNA molecule. The present invention further comprises the therapeutic and diagnostic use of the protein and/or DNA, e.g., a diagnostic kit for determining the presence and/or type of coagulase negative staphylococci and a vaccine composition, comprising the protein or DNA.

Abstract: The present invention relates to a novel enzyme (&agr;-GARE) which releases an amino acid residue having a glycated &agr;-amino group (&agr;-GA) from a glycated protein etc. and to bacterial strains producing the same. Examples of the bacterial strains include Corynebacterium ureolyticum KDK1002 (FERM P-17135) and Pseudomonas alcaligenes KDK1001 (FERM P-17133). The &agr;-GARE is contained in the culture supernatant of these strains and &agr;-GA can be released from a glycated peptide by using the same, as shown in FIG. 1.

Abstract: The present invention relates to live attenuated bacteria of the genus Actinobacillus pleuropneumoniae that have a mutation in an apxIV gene such that no functional ApxIV toxin can be produced. The invention also relates to methods for the production of such bacteria. Also vaccines comprising such bacteria and methods for the production of such vaccines are part of the invention. The invention further relates to subunit vaccines comprising an ApxIV toxin, to methods for the production of such vaccines and to methods for the protection of animals against infection with bacteria of the genus Actinobacillus pleuropneumoniae. In addition, the invention relates to the promotor of the apxIV gene. Finally, the invention relates to diagnostic test for the selective diagnosis of Actinobacillus pleuropneumoniae infections and to diagnostic tests discriminating between Actinobacillus pleuropneumoniae field strains and vaccine strains.

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 present invention relates to subtilase enzymes of the I-S1 and I-S2 sub-groups having an additional amino acid in the active site loop (c) region from positions 125 to 132. The variant subtilases of the present invention exhibit improved wash performance in a detergent in comparison to its parent enzyme.

Abstract: The present invention relates to novel subtilases having an improved wash performance on egg stains. The present invention also relates to isolated nucleic acid sequences encoding the subtilases, nucleic acid constructs, recombinant expression vectors, host cells comprising the nucleic acid construct, and methods for producing and using the subtilases of the invention. Further, the present invention relates to cleaning and detergent compositions comprising the subtilase enzymes of the invention as well as to use of such enzymes in detergent compositions and for removal of egg stains.

Abstract: The present invention relates to subtilase enzymes of the I-S1 and I-S2 sub-groups having an additional amino acid in the active site loop (b) region from positions 95 to 103. The variant subtilases of the present invention exhibit improved wash performance in a detergent in comparison to its parent enzyme.

Abstract: The present invention is a substantially purified sortase-transamidase enzyme from Gram-positive bacteria, such as Staphylococcus aureus.

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: An endopeptidase is described which shows specificity for cleavage of a LPXTG (SEQ ID NO: 1) motif found in the cell membranes of gram positive bacteria, having an apparent molecular weight of 14,000 daltons, having a pH optimum of about 7.5 to 10, being salt sensitive, being heavily glycosylated, rich in alanine, lacking aromatic amino acids, having a Km of 0.26 mM and having a backbone comprised of about 30% unknown amino acids, and containing carbohydrates that are essential for its activity. Methods of use and pharmaceutical compositions of this inhibitor are described.

Abstract: The present invention describes the identification of novel non-ribosomal peptide synthetases and associated biosynthetic genes from Streptomyces hygroscopicus. The present invention further provides methods for generating novel compounds, such as antibiotics, from these synthetases and associated genes.

Abstract: The present invention relates to the identification of novel cysteine proteases in Gram-positive microorganisms. The present invention provides the nuclei acid and amino acid sequences for the Bacillus subtilis cysteine proteases CP1, CP2 and CP3. The present invention also provides host cells having a mutation or deletion of part or all of the gene encoding CP1, CP2 or CP3. The present invention also provides host cells further comprising nucleic acid encoding desired heterologous proteins such as enzymes. The present invention also provides a cleaning composition comprising a cysteine protease of the present invention.

Abstract: An alkaline protease having the following properties; a gene encoding the same; a microorganism producing the same; and washing compositions containing the same; (i) acting over a broad pH value range of 4 to 13 and achieving, at pH 6 to 12, 80% or more the activity at the optimum pH value; (ii) when treated at 40° C. for 30 minutes, being stable over a pH value range of 6 to 11; (iii) having an isoelectric point of about 8.9 to 9.1; and (iv) having casein digesting activity that is not inhibited by oleic acid. The alkaline protease of the present invention is highly stable to various surface active agents and fatty acids, and exhibits high stability to oxidizing agents, and is therefore useful as an enzyme to be used in detergents for automatic dishwashers and laundry detergents, both containing bleaching components.

Abstract: This invention relates to the discovery that toxicity to mustard may be evaluated by diagnostic test means disclosed herein. Upon electrophoretic separation (sodium dodocyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)) of buffered extract of human skin cells (normal human epidermal keratinocytes (NHEK)) which had been exposed to mustard-type chemical compounds a band at approximately 50,000 to 80,000 daltons molecular weight was found. The protein band constitutes a biomarker.

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 present invention relates to the use of a subtilase variant for removal of egg stains from laundry or from hard surfaces, where the subtilase variant comprises at least one additional amino acid residue in the active site loop (b) region from position 95 to 103 (BASBPN numbering). These subtilase variants are useful exhibiting excellent or improved wash performance on egg stains when used in e.g. cleaning or detergent compositions, including automatic dishwash compositions. The present invention also relates to novel subtilase variants, to isolated DNA sequences encoding the variants, expression vectors, host cells, and methods for producing and using the variants of the invention. Further, the present invention relates to cleaning and detergent compositions comprising the variants of the invention.

Abstract: The present invention relates to the identification and use of a family of human complement C3-degrading proteinases expressed by S. pneumoniae. The proteinase has a molecular weight of about 24 kD to about 34 kD as determined on a 10% SDS polyacrylamide gel. A preferred proteinase of this invention includes the amino acid sequence of SEQ ID NO: 2.

Abstract: The structure and specificity of a recombinant &agr;2,3-sialyltransferase from Campylobacter spp., is disclosed. Also provided are methods for using the &agr;2,3-sialyltransferase in the production of desired carbohydrate structures and nucleic acids that encode the sialyltransferase.

Abstract: The present invention relates to enzymes produced by mutating the genes for a number of subtilases and expressing the mutated genes in suitable hosts are presented. The enzymes exhibit improved stability and/or improved wash performance in any detergent in comparison to their wild type parent enzymes. The enzymes are well-suited for use in any detergent and for some in especially liquid or solid shaped detergent compositions.

Abstract: Novel protease variants derived from the DNA sequences of naturally-occurring or recombinant non-human proteases are disclosed. The variant proteases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant protease to generate the substitution of a plurality of amino acid residues in the amino acid sequence of a precursor protease. Protease variants are provided that contain substitutions of the amino acids at one or more residue positions so that the substitution alters the charge at that position to make the charge more negative or less positive compared to a precursor protease and thus the protease variant is more effective in a low detergent concentration system than a precursor protease.

Abstract: The present invention relates to a novel microorganism Streptomyces megasporus SD5. The present invention also relates to a process for the isolation of said Streptomyces megasporus SD5. The invention also relates to a novel fibrinolytic enzyme actinokinase extracted from said microorganism and to a process for the extraction of said enzyme. In another aspect, the invention also pertains to a method for the treatment of thrombolytic disorders using said enzyme.

Abstract: Enzymes produced by mutating the genes for a number of subtilisin proteases and expressing the mutated genes in suitable hosts are presented. The enzymes exhibit improved wash performance in comparison to their wild type parent enzymes. The enzymes are well-suited for use in detergent compositions.

Abstract: Methods for controlling blood coagulation, and suitable pharmaceutical compositions that include a polypeptide that enhances the anticoagulation process (or inhibitors thereof for reversing the anticoagulation process) are provided.

Abstract: The present invention relates to polypeptide-polymer conjugates having added and/or removed one or more attachment groups for coupling polymeric molecules on the surface of the polypeptide structure, a method for preparing polypeptide-polymer conjugates of the invention, the use of said conjugates for reducing the immunogenicity and allergenicity and compositions comprising said conjugate.

Abstract: The present invention relates to cleaning compositions comprising a protease variant.

Abstract: A protease subtilase enzyme, characterized by an insertion in at least one active site loop. The enzymes exhibit improved wash performance in a detergent in comparison to its parent enzyme if it is a subtilase variant.

Abstract: The present invention relates to a novel improved protein mutant which produces low allergenic response in humans compared to the parent of that mutant. Specifically, the present invention comprises neutralizing or reducing the ability of T-cells to recognize epitopes and thus prevent sensitization of an individual to the protein.

Abstract: The present invention relates to variants of subtilisin-like proteases having decreased immunogenicity relative to their corresponding wild-type proteases. More particularly, the present invention relates to variants having a modified amino acid sequence of a wild-type amino acid sequence, wherein the modified amino acid sequence comprises a substitution of one or more epitope regions. The invention further relates to mutant genes encoding such variants and cleaning and personal care compositions comprising such variants.

Abstract: The present invention relates to variants of subtilisin-like proteases having decreased immunogenicity relative to their corresponding wild-type proteases. The present invention further relates to such variants additionally having one or more amino acid substitutions in one or more epitope regions or additionally having one or more stabilizing substitutions. The invention further relates to mutant genes encoding such variants and cleaning and personal care compositions comprising such variants.

Abstract: Novel carbonyl hydrolase variants derived from the DNA sequences of naturally-occurring or recombinant non-human carbonyl hydrolases are disclosed. The variant carbonyl hydrolases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant carbonyl hydrolase to generate the substitution of a plurality of amino acid residues in the amino acid sequence of a precursor carbonyl hydrolase. Such variant carbonyl hydrolases have properties which are different from those of the precursor hydrolase, such as altered proteolytic activity, altered stability, etc.

Abstract: Co-administration of a lysostaphin or other anti-staphylococcal agent which cleaves cross-links of peptidoglycans of staphylococci cell walls such as lysostaphin and an antibiotic effective against staphylococci due to antibiotic activity mediated by cell-wall activity is effective against staphylococcal infection, even staphylococci that may be resistant to one or other of lysostaphin or the cell-wall active antibiotic. Co-administration simultaneously suppresses the generation of antibiotic-resistant mutant strains. Effective cell-wall active antibiotics include &bgr;-lactams and glycopeptides.

Abstract: Novel protease variants derived from the DNA sequences of naturally-occurring or recombinant non-human proteases are disclosed. The variant proteases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant protease to generate the substitution of a plurality of amino acid residues in the amino acid sequence of a precursor protease. Such variant proteases have properties which are different from those of the precursor protease, such as altered wash performance.

Abstract: The invention relates to a process for obtaining L-dihydroorotic acid by chromatography on an anionic exchange material in a base water mixture under a pressure from about 1.1 MPa to about 40 MPa. The process can be used to investigate the in vitro and in vivo activity of N-(4-trifluoromethylphenyl)-5-methylisoxazole-4-carboxamide, N-(4-trifluoromethylphenyl)-2-cyano-3-hydroxycrontonamide and similar compounds.