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: To provide an Italian rye grass with excellent characteristics, and in particular, excellent insect resistance and disease resistance, an endophyte, which is a filamentous endophytic fungus living together with a wild plant occurring in nature, is isolated and artificially grown, and made to live symbiotically in Italian rye grass by inoculating and infecting the grass with the artificially grown endophyte.

Abstract: The present invention provides two novel human cathepsin proteins (HCPs) and polynucleotides encoding HCPs. The invention provides for genetically engineered expression vectors and host cells comprising the nucleic acid sequence encoding HCPs. The invention also provides for the production and use of antibodies to HCPs in pharmaceutical compositions for the treatment of disease processes that include cancers, inflammation, metastasis and peptide and proenzyme processing. In addition, the invention provides for the production and use of inhibitors of HSPs in pharmaceutical compositions for the treatment of diseases. The invention also describes diagnostic assays which utilize the polynucleotide to hybridize with the transcripts encoding HCPs. The invention also provides for the use of antisense molecules in pharmaceutical compositions as a therapeutics in cancers, inflammation, metastasis and peptide and proenzyme processing.

Abstract: The present invention provides a method for producing a secretable polypeptide in a host cell, comprising overexpressing a peptidyl proly cis-trans isomerase in the host cell, thereby increasing the yield of the secreted polypeptide, and a novel peptidyl proly cis-trans isomerase from A. niger useful in such a method.

Abstract: The invention pertains to the field of biology, and more particularly, to therapeutic chemistry, and discloses pharmaceutical compositions containing, as an active ingredient, an enzyme or an enzyme complex containing said enzyme, obtained by cultivating Streptomyces fradiae under conditions defined in French patent 2.600.340, as a mixture or in combination with a physiologically acceptable, non-toxic and inert carrier or vector. Said pharmaceutical compositions are useful for preventing or treating degenerative diseases, particularly those related to intestinal dysfunction.

Abstract: The invention includes novel fusion DNA sequences encoding fusion polypeptides which when expressed in a filamentous fungus result in the expression of fusion polypeptides which when secreted result in increased levels of secretion of the desired polypeptide as compared to the expression and secretion of such polypeptides from filamentous fungi transformed with previously used DNA sequences. The fusion DNA sequences comprise from the 5′ terminus four DNA sequences which encode a fusion polypeptide comprising, from the amino to carbonyl-terminus, first, second, third and fourth amino acid sequences. The first DNA sequence encodes a signal peptide functional as a secretory sequence in a first filamentous fungus. The second DNA sequence encodes a secreted polypeptide or portion thereof which is normally secreted from the same filamentous fungus or a second filamentous fungus. The third DNA sequence encodes a cleavable linker polypeptide while the fourth DNA sequence encodes a desired polypeptide.

Abstract: Novel vectors are disclosed for expressing and secreting heterologous polypeptides from filamentous fungi. Such vectors are used in novel processes to express and secrete such heterologous polypeptides. The vectors used for transforming a filamentous fungus to express and secrete a heterologous polypeptide include a DNA sequence encoding a heterologous polypeptide and a DNA sequence encoding a signal sequence which is functional in a secretory system in a given filamentous fungus and which is operably linked to the sequence encoding the heterologous polypeptide. Such signal sequences may be the signal sequence normally associated with the heterologous polypeptides or may be derived from other sources. The vector may also contain DNA sequences encoding a promoter sequence which is functionally recognized by the filamentous fungus and which is operably linked to the DNA sequence encoding the signal sequence.

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: The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N-or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.

Abstract: Novel vectors are disclosed for expressing and secreting heterologous polypeptides from filamentous fungi. Such vectors are used in novel processes to express and secrete such heterologous polypeptides. The vectors used for transforming a filamentous fungus to express and secrete a heterologous polypeptide include a DNA sequence encoding a heterologous polypeptide and a DNA sequence encoding a signal sequence which is functional in a secretory system in a given filamentous fungus and which is operably linked to the sequence encoding the heterologous polypeptide. Such signal sequences may be the signal sequence normally associated with the heterologous polypeptides or may be derived from other sources. The vector may also contain DNA sequences encoding a promoter sequence which is functionally recognized by the filamentous fungus and which is operably linked to the DNA sequence encoding the signal sequence.

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: A method of producing a milk clotting enzyme including the steps of (a) fermenting a strain of Rhizomucor miehei or Aspergillus oryzae to form a fermentation product having a glycoslated Rhizomucor miehei aspartic protease and other proteins, and (b) subjecting a quantity of the fermentation product to a deglycosylating treatment to form a coagulant preparation having an at least partly deglycoslated aspartic protease and the other proteins. The at least partly deglycosylated protease has a milk clotting activity that is at least 10% higher than a milk clotting activity of the glycosylated aspartic protease.

Abstract: The present invention provides a fungal lichenase, i.e., an endo-1,3-1,4-.beta.-D-glucanohydrolase, its coding sequence, recombinant DNA molecules comprising the lichenase coding sequences, recombinant host cells and methods for producing same. The present lichenase is from Orpinomyces PC-2.

Abstract: The present invention has for an object a koji mold which expresses at least 2 times more endo- and exo-peptidases than the wild type strain of Aspergillus oryzae CNCM I-1882, and especially at least 30 mU of endopeptidase activity, at least 30 mU of leucine-amino-peptidase and at least 10 mU of prolyldipeptidyl-peptidase activity per ml of supernatant when grown in a minimal medium containing 0.2% soy bean proteins. The invention also provides a DNA-binding protein of Aspergillus oryzae (AREA) having at least the amino-acid sequence from amino-acid 1 to amino-acid 731 of SEQ ID NO:2 or functional derivatives thereof. The invention also provides a DNA molecule that comprises an areA gene encoding the DNA-binding protein according to the invention.

Abstract: The present invention relates to a new cassette for the expression of an endothiapepsin precursor in Cryphonectria parasitica, to a strain of this species transformed with this cassette, to a process for preparing endothiapepsin using this strain and also to a process for preparing such a strain devoid of a dominant selection marker.

Abstract: An isolated and purified enzyme exhibiting protease activity at a pH of 4-7 which exhibits protease activity in 5% hydrogen peroxide and which is encoded by a DNA sequence which hybridizes to a DNA sequence depicted in SEQ ID NO. 1 or 2.

Abstract: A method for transamidating a peptide substrate having a P.sub.1 amino acid residue with a positively charged side chain. According to the invention, carboxypeptidase Y is modified to substitute at least one amino acid having a negatively charged side chain in an S.sub.1 subsite. Additionally, the modified carboxypeptidase Y can include substituted amino acid residues in an S.sub.1 ', S.sub.2 and/or S.sub.3 subsite to accommodate a specific peptide substrate.

Abstract: The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N- or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.

Abstract: Disclosed are methods that can be used to (1) measure the level of polysaccharide in a sample; (2) measure the ability of a compound to degrade a polysaccharide; (3) measure the ability of a compound to modulate polysaccharide synthesis; and (4) identify or distinguish a polysaccharide, and hence organism, for diagnostic purposes in clinical medicine or research. The invention stems from Applicant's discovery that polysaccharides have multiple binding sites for polysaccharide binding moieties (PBM, e.g., wheat germ agglutinin (WGA)). In each method, one PBM links the polysaccharide to a substrate, and a tagged PBM is used to detect the polysaccharide. All of these methods can be carried out rapidly and quickly in the wells of a microtiter plate, thus permitting high through-put screening of samples or test compounds.

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: A DNA construct comprising a DNA sequence encoding an enzyme exhibiting protease activity, which DNA sequence comprises the DNA sequence shown in SEQ ID No. 1 or 2 or an analog of any of these sequences being at least 80% homologous to the DNA sequence shown in SEQ ID No. 1 or 2. The proteases encoded by the DNA sequences have an acid pH optimum.

Abstract: The present invention relates to a novel metalloprotease obtainable from a fungus having increased proteolytic activity. Additionally, the invention related to isolated nucleic acid fragments encoding said metalloprotease as well as vectors, DNA constructs, and recombinant host cells comprising said nucleic acid fragments.

Abstract: The present invention is directed to an improved process for preparing in high yield an obesity protein analog using a dipeptidylaminopeptidase isolated from the slime mold, Dictyostelium discoideum.

Abstract: The present invention relates to a novel metalloprotease obtainable from a fungus having increased proteolytic activity. Additionally, the invention related to isolated nucleic acid fragments encoding said metalloprotease as well as vectors, DNA constructs, and recombinant host cells comprising said nucleic acid fragments.

Abstract: The invention relates to serine protease variants derived from precursor serine proteases via recombinant and/or chemical methods to form protease variants having improved peptide ligase activity. The invention also includes novel ligation substrates which in combination with the serine protease variants and a second ligation substrate are capable of forming a ligation product. The invention also relates to methods for forming such ligation products and the products formed thereby.

Abstract: Beneficial endophytes which live within certain plants are known to provide desirable, cost-effective biological insect control. Many naturally occurring grasses host symbiotic endophytic fungi. However, beneficial endophytes have never been found in several species of turf grass, including species of bentgrasses and Kentucky bluegrasses, two commercially important turf grasses which are used extensively on golf courses and for lawn turfs. The invention of this application relates to new methods of inoculating plant tissues which allow the development of endophyte-enhanced varieties of turfgrasses, and to the turf grass varieties produced using the methods.

Abstract: The present invention is related to a process for producing an active enzyme comprising fermenting the proform of the active enzyme in the present of a proteolytic enzyme different from the active enzyme and capable of converting the proenzyme into an active enzyme as well as to host cells, recombinant expression vectors and host cells suitable for use in the process.

Abstract: An active recombinant trypsin-like protease enzyme comprising the amino acid residues 25-224 of the amino acid sequence of SEQ ID NO:2, as well as a DNA construct encoding the enzyme and comprising the sequence of SEQ ID NO:1. The enzyme may be used as a detergent enzyme.

Abstract: The present invention relates to a novel metalloprotease obtainable from a fungus having increased proteolytic activity. Additionally, the invention related to isolated nucleic acid fragments encoding said metalloprotease as well as vectors, DNA constructs, and recombinant host cells comprising said nucleic acid fragments.

Abstract: A laundry detergent for washing fabrics composed of proteinogenic fibers is comprised of at least one surfactant and a proteolytically active amount of a protease having a keratinase/caseinase activity ratio of less than about 0.80.

Abstract: The invention herein provides bleaching compositions comprising a protease enzyme which is a carbonyl hydrolase variant having an amino acid sequence not found in nature, which is derived by replacement of a plurality of amino acid residues of a precursor carbonyl hydrolase with different amino acids, wherein said plurality of amino acid residues replaced in the precursor enzyme correspond to position +76 in combination with one or more of the following residues: +99, +101, +103, +104, +107, +123, +27, +105, +109, +126, +128, +135, +156, +166, +195, +197, +204, +206, +210, +216, +217, +218, +222, +260, +265, and/or +274, where the numbered positions corresponds to naturally-occurring subtilisin from Bacillus amyloliquefaciens or to equivalent amino acid residues in other carbonyl hydrolases or subtilisins (such as Bacillus lentus subtilisin) and a bleaching agent.

Abstract: The invention concerns novel DNA molecules encoding a modified, endoplasmic reticulum-located "dibasic processing endoprotease" and the use of said endoplasmic reticulum-located "dibasic processing endoprotease" for the correct processing of heterologous polypeptides in transformed hosts.

Abstract: An enzyme feed additive is provided comprising a xylanase, a protease, and optionally a .beta.-glucanase. The ratio of the units of xylanase activity per unit amount of the feed additive to the units of .beta.-glucanase activity per same unit amount of the feed additive is 1:0-0.25.Preferably, the xylanase is the low pI xylanase and/or the high pI xylanase obtained from Trichoderma longibrachiatum.Preferably, the protease is a mutant subtilisin comprising a substitution at the amino acid residue position equivalent to tyr+217 of Bacillus amyloliquefaciens subtilisin with leucine.

Abstract: A method for removing dipeptides from the amino terminus of precursor polypeptides to produce a polypeptide product is presented which comprises contacting the precursor polypeptide for sufficient time to remove the dipeptide with an immobilized dipeptidylaminopeptidase (dDAP) from the slime mold, Dictyostelium discoideum, which has a mass of about 225 kilodaltons and a pH optimum of about 3.5. The precursor polypeptides may be made recombinantly and may be analogs of naturally occurring polypeptides.

Abstract: A dipeptidylaminopeptidase (dDAP enzyme) and a method for isolating it from the culture medium of the cellular slime mold, Dictyostelium discoideum have been presented. The isolated dDAP enzyme has a pH optimum of about 3.5 and a mass of about 225 kDA. The dDAP enzyme has an activity which is somewhat similar to both DAP-I and DAP-III from this organism. Methods for using the dDAP enzyme to remove dipeptides from the N-terminus of recombinantly produced precursor proteins or peptides are also presented.

Abstract: A method for removing dipeptides from the amino terminus of precursor polypeptides to produce a polypeptide product is presented which comprises contacting the precursor polypeptide for sufficient time to remove the dipeptide with a dipeptidylaminopeptidase (dDAP) from the slime mold, Dictyostelium descoideum, which has a mass of about 225 kilodaltons and a pH optimum of about 3.5. The precursor polypeptides may be made recombinantly and may be analogs of naturally occurring polypeptides.

Abstract: The present invention is directed to a method for screening samples for the identification of agents exhibiting potential fungicidal and insecticidal activity for a wide variety of agricultural, medical and pharmaceutical uses. The method utilizes cells that comprise a plasmid-born CTS gene of Saccharomyces cerevisiae, which allows for over expression of chitinase.

Abstract: Nonaqueous gelled automatic dishwashing compositions containing a mixture of a protease enzyme and an amylase enzyme have been found to be very useful in the removal of protein and carbohydrate soils from dishware at operating temperatures of 100.degree. F. to 140.degree. F.

Abstract: The invention concerns novel DNA molecules encoding a modified, endoplasmic reticulum-located "dibasic processing endoprotease" and the use of said endoplasmic reticulum-located "dibasic processing endoprotease" for the correct processing of heterologous polypeptides in transformed hosts.

Abstract: The disclosure relates to a generic class of ubiquitin-specific proteases which specifically cleave at the C-terminus of the ubiquitin moiety in a ubiquitin fusion protein irrespective of the size of the ubiquitin fusion protein. More specifically, the disclosure relates to ubiquitin-specific proteases of this class which have been isolated from a cell. The disclosure also relates to isolated DNA sequences encoding the proteases of this class.

Abstract: A protease obtainable from Dendryphiella arenaria DSM 6260 or Dendryphiella salina DSM 6332 is disclosed. The protease has a pl of approximately 9.1 as determined by isoelectric focusing, a molecular weight of approximately 18-20 kD as determined by SDS-PAGE, is active towards haemoglobin, casein, skimmed milk, and Suc-Ala-Ala-Pro-Phe-pNA and has immunochemical properties identical to those of a protease obtained from Dendryphiella arenaria DSM 6260 or Dendryphiella salina DSM 6332. A process for its production, use in a detergent composition and as a detergent additive are disclosed.

Abstract: A crude membrane fraction of leaf sheath tissue of a grass endophytically infected by A. typhinum is produced. The extract exhibits endoproteolytic activity in the presence of detergent or after methanol precipitation. This activity is optimal at 35.degree.-40.degree. C. and pH 10-11 and is exhibited only when a reductant is present. The activity is associated with a first gel electrophoresis band having apparent molecular weight of 205,000 daltons when said extract is electrophoresed without prior boiling; however, when said extract is boiled prior to electrophoresis, there results a band having apparent molecular weight of 34,000 daltons. This band gives rise to polyclonal rabbit antibodies which are not cross react with proteinase K. The preferred source of Acremonium typhinum is ATCC 74228.

Abstract: A high density, enzyme-containing powder detergent composition including a combination of alkaline proteases for improved cleaning characteristics and a combination of different density sodium carbonates for improved dispensing characteristics.

Abstract: This invention is in the field of detergent proteases derived from strains of a new Bacillus sp. More specifically, the invention is directed towards a protease derived from a strain of Bacillus sp. TY 145. Moreover, the invention is directed towards a process for the preparation of the protease the use of the protease enzyme, and detergent composition comprising the protease of the invention.

Abstract: An enzyme produced by a microorganism belonging to the genus Myrothecium hydrolyzes .gamma.-polyglutamic acid with an endo action to produce oligoglutamic acid consisting of 2 to 4 glutamic acid residues.

Abstract: A mutant fungus strain which produces a protease with low thermostability and low productivity was made from protease producing fungi and the gene coding for mutant enzyme is isolated from the mutant strain. A promoter which can function in yeast is ligated to the gene and inserted into a plasmid replicable in yeast, and the resulting plasmid is introduced into yeast. The yeast is cultured, thereby producing the mutant enzyme. Furthermore, a gene expressing an enzyme with a far lower thermostability is prepared by site-directed mutagenesis, which is introduced in yeast, thereby producing an enzyme with more distinctively reduced thermostability.

Abstract: This invention relates to a method of selectively degrading .beta.-lactoglobulin contained in cow's milk-serum protein by using a specific enzyme capable of selectively degrading .beta.-lactoglobulin.

Abstract: A method of producing an angiotensin converting enzyme inhibitor-containing composition, which is of value as an antihypertensive agent or diet, from natural resources. According to the method, proteins are hydrolyzed with a protease elaborated by a microorganism of the genus Bacillus, Aspergillus or Rhizopus or a protease of papaya origin.

Abstract: An endoprotease preparation, comprising an isolated endoprotease is disclosed. The endoprotease is a serine protease, shows immunochemical idenity to a protease derived from Fusarium oxysporum DSM 2672, hydrolyzes the oxidized beta-chain of bovine insulin at the peptide bonds Arg (22)-Gly (23) and Lys (29)-Ala(30), has optimum activity towards casein in the pH range of 8.5-11.0 with nearly constant activity in the pH range, has optimum activity at a temperature of about 45.degree. C., and has an isolectric point of about 9.0-10.0. The preferred microorganism that the enzyme is isolated from is Fusarium oxysporum DSM 2672.

Abstract: This disclosure relates to a novel class of serine proteases isolated from a culture medium of fungus Tritirachium album. The serine proteases disclosed have a high degree of stability in detergent formulations.In addition, this disclosure relates to a process for producing such serine proteases using recombinant techniques.