Abstract: Methods for identifying organisms capable of degrading fumonisin. Fumonisin can be incorporated into culture medium for selection of organisms resistant to fumonisin and/or capable of growing on fumonisin as a sole carbon source. Using this method, several organisms have been identified. These organisms can be used to isolate the enzymes and the genes responsible for conferring fumonisin-resistance. The gene can be cloned and inserted into a suitable expression vector so that the protein can be further characterized. Additionally, the DNA encoding for fumonisin degrading enzymes can be used to transform plant cells normally susceptible to Fusarium or other toxin-producing fungus infection. Plants can be regenerated from the transformed plant cells. In this way, a transgenic plant can be produced with the capability of degrading fumonisin, as well as with the capability of producing the degrading enzymes.

Abstract: The present invention provides a transgenic animal assay system which provides a model system for testing for, and treatment of, cholinergic deficits and imbalances in mammals such as cognitive functioning in Alzheimer's patients, certain types of retinal photoreceptor degeneration, hematopoietic disorders, and screening for and susceptibility to anti-cholinesterase compounds. The transgenic animals and progeny thereof are transformed with a recombinant expression vector of the present invention. The recombinant expression vector comprises a DNA sequence encoding a heterologous cholinesterase (ChE) enzyme and promoter.

Abstract: The present invention is directed toward efficient, high-yield processes for making ascorbic acid, 2-keto-L-gulonic acid, and esters of 2-keto-L-gulonic acid. The processes comprise reacting the appropriate starting materials with a hydrolase enzyme catalyst such as a protease, an esterase, a lipase or an amidase.

Abstract: The present invention discloses a process for improving the efficiency of feed utilization and/or for promoting the growth of animals in which an animal is fed a diet which comprises a composition comprising feed substance and a ready for use phospholipase additive. Preferably said composition also comprises at least one phospholipid. Said compositions are used to improve fat digestibility and to promote growth of the animal. The phospholipid is preferably lecithin and the preferred phospholipase is a mammalian phospholipase A2. In a preferred embodiment the phospholipase is produced using recombinant DNA technology to express the enzyme in a suitable host such as a microorganism or a transgenic plant.

Abstract: Methods and DNA constructs are provided for the expression of a fungal acetyl xylan esterase gene in microbial hosts. A purified fungal acetyl xylan esterase is obtained which is suited for the use as an accessory enzyme in the degradation of acetylated xylans.

Abstract: The invention provides a new human lysophospholipase (IHLP) and polynucleotides which identify and encode IHLP. The invention also provides expression vectors, host cells, agonists, antibodies and antagonists. The invention also provides methods for treating inflammation and disorders associated with expression of IHLP.

Abstract: Disclosed are a thermophilic phospholipase having an optimum temperature range of 95 to 105.degree. C. which is useful in high-temperature degumming processes in oil refining process and high-temperature processing of phospholipids; and a method for producing a thermophilic phospholipase comprising culturing a microorganism capable of producing the phospholipase in a culture medium and collecting the phospholipase from the resultant culture.

Abstract: A method of modifying esterases by substitution with histadine of at least one amino acid within 6 .ANG. of an active site serine provides esterases useful for detoxifying organophasphates.

Abstract: Two previously undescribed human cdc25 genes, designated cdc25 A and cdc25 B, which have been shown to have an endogenous tyrosine phosphatase activity that can be specifically activated by B-type cyclin, in the complete absence of cdc2 are described. As a result of this work, new approaches to regulating the cell cycle in eukaryotic cells and, particularly, to regulating the activity of tyrosine specific phosphatases which play a key role in the cell cycle are available. Applicant's invention relates to methods of regulating the cell cycle and, specifically, to regulating activation of cdc2-kinase, through alteration of the activity and/or levels of tyrosine phosphatases or through alteration of the interaction of components of MPF. The present invention also relates to agents or compositions useful in the method of regulating (inhibiting or enhancing) the cell cycle.

Abstract: A process for preparing acrylic or methacrylic acid amides functionalized with a cationizable tertiary amine, by aminolysis of acrylic or methacrylic acid esters with the aid of enzymes.

Abstract: The present invention relates to polyhydroxyalkanoate depolymerase having a N-terminal fragment of the amino acid sequence of SEQ ID NO: 1 and having a molecular weight of about 33,000 as determined by SDS polyacrylamide gel electrophoresis; a process for producing polyhydroxyalkanoate depolymerase, comprising culturing in a medium a microorganism which belongs to the genus Corynebacterium and has the ability to produce the polyhydroxyalkanoate depolymerase and recovering the polyhydroxyalkanoate depolymerase from the resulting culture; and a process for producing polyhydroxyalkanoate depolymerase, comprising culturing in a medium a transformant transformed with a recombinant vector containing a gene of the polyhydroxyalkanoate depolymerase and recovering the polyhydroxyalkanoate depolymerase from the resulting culture. According to the present invention, there can be provided novel PHA depolymerase with the activity of decomposing .omega.

Abstract: Optically active 2-substituted tetrahydropyran-4-ols or esters thereof may be prepared using esterases or hydrolases from the corresponding racemic mixtures of esters or alcohols. This provides a route to the corresponding optically active ketones.The racemic mixtures are preferably in the cis-form. such mixtures may be produced by reacting but-3-ene-1-ol with an aldehyde in the presence of an acid.

Abstract: A process for resolving racemic diesters of (R,S)(.+-.)-5-hydroxy-3-(4'-hydroxyphenyl)-1,1,3-trimethylindane is disclosed. The process utilizes a microbial enzyme derived from Chromobacterium viscosum to catalyze the enantioselective and regioselective hydrolysis of the (S)(-)-enantiomer of the racemic mixture to its corresponding monoester at a faster rate than the (R)(+)-enantiomer. A substantially pure (S)(-)-indane monoester is thereby formed, while the (R)(+)-indane diester remains unreacted. The (S)(-)-monoester and the (R)(+)-diester can then be hydrolyzed using conventional techniques to form optically active (R)(+)- and (S)(-)-5-hydroxy-3-(4'-hydroxyphenyl)-1,1,3-trimethylindane enantiomers for use as monomers in the synthesis of chiral polymers.

Abstract: The invention relates to the gene encoding the enzyme cellobiohydrolase-1. Specifically, the invention concerns the elucidation of the regulatable promoter sequence of said gene and the subsequent genetic manipulation of said sequence so as to combine it with DNA sequence structure of a heterologous peptide in order to provide for selective expression of heterologous peptide in accordance with the expression features of the promoter.

Abstract: A method for isolating and identifying modified para-nitrobenzyl esterases which exhibit improved thermal stability relative to naturally occurring para-nitrobenzyl esterase. The method involves preparing a library of modified para-nitrobenzyl esterase nucleic acid segments (genes) which have nucleotide sequences that differ from the nucleic acid segment which encodes for naturally occurring para-nitrobenzyl esterase. The library of modified para-nitrobenzyl nucleic acid segments is expressed to provide a plurality of modified enzymes. The clones expressing modified enzymes are then screened to identify which enzymes retain esterase activity after heat treatment at elevated temperature. Specific modified para-nitrobenzyl esterases are disclosed which have improved thermal stability and/or ester hydrolysis activity in aqueous or aqueous-organic media relative to the thermal stability and/or ester hydrolysis activity of unmodified naturally occurring para-nitrobenzyl esterase.

Abstract: Esterase enzymes derived from various Staphylothermus, Pyrodictium, Archaeoglobus, Aquifex, M11TL, Thermococcus, Teredinibacter and Sulfolobus organisms are disclosed. The enzymes are produced from native or recombinant host cells and can be utilized in the pharmaceutical, agricultural and other industries.

Abstract: Provided herein are signal peptides, and constructs and vectors having signal peptides, as well as methods of using signal peptides to deliver proteins into secretory material. The invention facilitates the purification of recombinant proteins. Also provided are a new class of proteins called nectarins, from which the signal peptides may be derived. Additionally, the nectarins possess oxalate oxidase activity and have utility as agents for treating diseases or conditions relating to abnormal oxalate deposition or metabolism or as diagnostics for measuring oxalate concentrations in specimens.

Abstract: The regioselective and chemoselective hydrolysis of an .alpha.-ester group of an amino acid diester using pig liver esterase enzyme (PLE) is disclosed. The amino acid diesters may be either N-protected or unprotected and the diester groups may be the same or different. In particular, the preparation of a number of .gamma.-ester glutamates and .beta.-ester aspartates are provided.

Abstract: A process for the microbiological or enzymatic hydrolytic resolution of racemic trans-2-(alkoxycarbonylethyl) lactams of the formula I: ##STR1## wherein R is C.sub.1 -C.sub.7 alkyl, 2,2,2-trifluoroethyl or methoxyethoxyethyl and R.sup.1 is hydrogen or a protecting group is disclosed, whereby an optically enriched compound of the formula Ib or IIa: ##STR2## is obtained.

Abstract: The invention provides two novel members of the cytochrome P450 2C subfamily of enzymes, designated 2C18 and 2C19. DNA segments encoding these enzymes are also provided. The 2C19 polypeptide represents the principal human determinant of human S-mephenytoin 4'-hydroxylase activity. The invention also provides methods of identifying drugs metabolized by S-mephenytoin 4'-hydroxylase activity. Drugs shown to be metabolized by this activity should in general not be administered to individuals having, or belong to an ethnic group at risk of, a polymorphic deficiency in S-mephenytoin 4'-hydroxylase activity. The invention also provides methods of diagnosing individuals having a polymorphic deficiency.

Abstract: A method for isolating and identifying modified para-nitrobenzyl esterases which exhibit improved stability and/or esterase hydrolysis activity toward selected substrates and under selected reaction conditions relative to the unmodified para-nitrobenzyl esterase. The method involves preparing a library of modified para-nitrobenzyl esterase nucleic acid segments (genes) which have nucleotide sequences that differ from the nucleic acid segment which encodes for unmodified para-nitrobenzyl esterase. The library of modified para-nitrobenzyl nucleic acid segments is expressed to provide a plurality of modified enzymes. The clones expressing modified enzymes are then screened to identify which enzymes have improved esterase activity by measuring the ability of the enzymes to hydrolyze the selected substrate under the selected reaction conditions.

Abstract: A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent .beta.-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.

Abstract: The instant invention provides nucleic acid molecules encoding cephalosporin esterase from Rhodosporidium toruloides. In addition, the invention provides isolated cephalosporin esterase, expression vectors, host cells, and methods of producing cephalosporin esterase.

Abstract: The present invention provides novel aerobic, Gram-positive alkaliphilic bacteria which have been isolated from in and around alkaline soda lakes. These alkaliphiles have been analyzed according to the principles of numerical taxonomy with respect to each other and also to a collection of known bacteria. In addition, these bacterial taxa are further circumscribed by an analysis of the lipid components which serve as chemotaxonomic markers. The alkaliphiles of the present invention produce alkalitolerant enzymes which are capable of performing their functions at high pH which makes them uniquely suited for applications requiring such extreme conditions.

Abstract: An oxidized phospholipid degrading enzyme is provided, which plays an important role in the oxygen stress preventive mechanism in organisms. The enzyme hydrolyzes a 1-acyl-2-.omega.-carboxyfatty acid acyl-3-phosphatidylcholine as a substrate at the 2-ester bond thereof to form 1-acyl-2-lyso-3-phosphatidylcholine. The activity of the enzyme is slightly enhanced by 4 mM calcium chloride. The molecular mass of the enzyme is 95.+-.5 kDa (by gel filtration). The enzyme is composed of three subunits whose molecular masses have been found to be 29 kDa, 30 kDa and 45 kDa, respectively, by SDS-polyacrylamide electrophoresis. A gene coding the enzyme is also provided. This gene is important for the synthesis of the enzyme by genetic engineering.

Abstract: The lipopolysaccharide of bacteria associated with chronic inflammatory diseases is unable to induce expression of leukocyte adhesion molecules, or selectins, on endothelial cells, and is also capable of inhibiting the induction of selectin expression by bacteria normally associated with acute endotoxin disease. New approaches to treatment of these diseases, and the diagnosis of susceptibility to chronic bacterial-associated inflammatory diseases, are provided.

Abstract: An isolated and purified acetyl esterase with activity towards acetylated xylan and acetylated mannan obtained from Aspergillus aculeatus and having amino acid sequence that comprises Seq ID No 1. An enzyme can be used for the modification of plant cell wall components.

Abstract: The present invention is directed toward efficient, high-yield processes for making ascorbic acid, 2-keto-L-gulonic acid, and esters of 2-keto-L-gulonic acid. The processes comprise reacting the appropriate starting materials with a hydrolase enzyme catalyst such as a protease, an esterase, a lipase or an amidase.

Abstract: The present invention is directed to a process for improving the property of proanthocyanidins comprising contacting the proanthocyanidin containing solution with tannase and a process for increasing the yield of proanthocyanidins comprising contacting the proanthocyanidin containing solution with tannase in the extraction of the proanthocyanidins.

Abstract: A process is presented for producing a stable carboxylesterase in dry form wherein a saccharide selected from maltose, maltotriose, maltotetrose, maltopentaose, maltohexaose, maltoheptaose and maltitol is added to the solution of carboxylesterase to be spray dried. The dry enzyme composition is added to food and drink (e.g., bread, brewed seasonings, liquors, processed meat products) during the production process to thereby impart thereto a good aroma (ester aromas).

Abstract: A method for genetically engineering cells to produce soluble and secretable Golgi processing enzymes instead of naturally occurring membrane-bound enzymes. Cells are genetically engineered to express glycosyltransferases which lack both a membrane anchor and a retention signal. The resulting altered enzyme becomes soluble and secretable by the cell without losing its catalytic activity. Secretion of the soluble glycosyltransferase by the cell provides for increased production and simplified recovery of glycosyltransferase.

Abstract: The regioselective and chemoselective hydrolysis of an .alpha.-ester group of an amino acid diester using pig liver esterase enzyme (PLE) is disclosed. The amino acid diesters may be either N-protected or unprotected and the diester groups may be the same or different. In particular, the preparation of a number of .gamma.-ester glutamates and .beta.-ester aspartates are provided.

Abstract: Two previously undescribed human cdc25 genes, designated cdc25 A and cdc25 B, which have been shown to have an endogenous tyrosine phosphatase activity that can be specifically activated by B-type cyclin, in the complete absence of cdc2 are described. As a result of this work, new approaches to regulating the cell cycle in eukaryotic cells and, particularly, to regulating the activity of tyrosine specific phosphatases which play a key role in the cell cycle are available. Applicant's invention relates to methods of regulating the cell cycle and, specifically, to regulating activation of cdc2-kinase, through alteration of the activity and/or levels of tyrosine phosphatases or through alteration of the interaction of components of MPF. The present invention also relates to agents or compositions useful in the method of regulating (inhibiting or enhancing) the cell cycle.

Abstract: Methods and DNA constructs are provided for the expression of a fungal acetyl xylan esterase gene in microbial hosts. A purified fungal acetyl xylan esterase is obtained which is suited for the use as an accessory enzyme in the degradation of acetylated xylans.

Abstract: Two previously undescribed human cdc25 genes, designated cdc25 A and cdc25 B, which have been shown to have an endogenous tyrosine phosphatase activity that can be specifically activated by B-type cyclin, in the complete absence of cdc2 are described. As a result of this work, new approaches to regulating the cell cycle in eukaryotic cells and, particularly, to regulating the activity of tyrosine specific phosphatases which play a key role in the cell cycle are available. Applicant's invention relates to methods of regulating the cell cycle and, specifically, to regulating activation of cdc2-kinase, through alteration of the activity and/or levels of tyrosine phosphatases or through alteration of the interaction of components of MPF. The present invention also relates to agents or compositions useful in the method of regulating (inhibiting or enhancing) the cell cycle.

Abstract: A process for efficiently producing (S,S)-2-alkoxycyclohexanols in a single step by using (.+-.)-trans-2-alkoxycyclohexanols which are inexpensive and can be easily obtained. The process comprises treating a (.+-.)-trans-2-alkoxycyclohexanol with a hydrolase originating in a microorganism and being capable of esterifying stereospecifically the R-isomer in the presence of an acyl donor under such conditions that no hydrolysis occurs substantially to thereby give (S,S)-2-alkoxycyclohexanols and (R,R)-2-alkoxycyclohexanol carboxylate and then taking up the (S,S)-2-alkoxycyclohexanols.

Abstract: A cloned DNA encoding phospholipase D originated from a plant and a cloned DNA which regulates expression of phospholipase D gene originated from a plant are disclosed.

Abstract: A method for isolating and identifying modified para-nitrobenzyl esterases which exhibit improved stability and/or esterase hydrolysis activity toward selected substrates and under selected reaction conditions relative to the unmodified para-nitrobenzyl esterase. The method involves preparing a library of modified para-nitrobenzyl esterase nucleic acid segments (genes) which have nucleotide sequences that differ from the nucleic acid segment which encodes for unmodified para-nitrobenzyl esterase. The library of modified para-nitrobenzyl nucleic acid segments is expressed to provide a plurality of modified enzymes. The clones expressing modified enzymes are then screened to identify which enzymes have improved esterase activity by measuring the ability of the enzymes to hydrolyze the selected substrate under the selected reaction conditions.

Abstract: A process for the production of monic acids which comprises contacting a corresponding pseudomonic acid or an ester thereof with a hydrolase enzyme from a suitable microorganism, in particular Streptomyces sp.

Abstract: Methods for identifying organisms capable of degrading fumonisin. Fumonisin can be incorporated into culture medium for selection of organisms resistant to fumonisin. Using this method, several organisms have been identified. These organisms can be used to isolate the enzyme and the gene responsible for conferring fumonisin-resistance. The gene can be cloned and inserted into a suitable expression vector so that the protein can be further characterized. Additionally, the DNA encoding for fumonisin-resistance can be used to transform plant cells normally susceptible to Fusarium infection. Plants can be regenerated from the transformed plant cells. In this way, a transgenic plant can be produced with the capability of degrading fumonisin.

Abstract: The individual enantiomers of the compounds (Ia), (Ib), (IIa) or (IIb), optionally substituted by non-interfering substituent(s). The novel enantiomers can be obtained by biotransformation. The (Ia) or (IIa) compounds can be used for the synthesis of chiral carbocyclic nucleosides.

Abstract: An enzyme exhibiting pectin methyl esterase activity, wherein the enzyme a) is derived from Aspergillus aculeatus; b) is encoded by Seq ID No:1; or c) has the amino acid sequence of Seq ID No:2 or a sequence which is at least 95% homologous thereto. An enzyme can be used for the modification of plant cell wall components.

Abstract: A norbornane type ester hydrolase that enantio-selectively hydrolyzes a (.+-.) -exo-norbornane type ester represented by Formula I is provided: ##STR1## wherein R is acyl, and A and B are hydrogens, respectively, or where A and B are absent, resulting in a carbon-carbon double bond between the carbons to which A and B are attached in Formula I. The norbornane type ester hydrolase has an optimal pH of approximately 8 and a stable pH range of approximately 6 to 8.

Abstract: Methods and DNA constructs are provided for the expression of a fungal acetyl xylan esterase gene in microbial hosts. A purified fungal acetyl xylan esterase is obtained which is suited for the use as an accessory enzyme in the degradation of acetylated xylans.

Abstract: Fatty acid esters of alkyl glycosides are enzymatically prepared by forming a micro-emulsion from a fatty acid, an alkyl glycoside and a nonionic or anionic surface active agent. The viscosity of the micro-emulsion is adjusted with tertiary butanol 2-methyl-1-butanol acetone or 2-butanone. After the enzymatic esterification has been effected, the reaction mixture is subjected to pervaporation.

Abstract: A gene activation/inactivation and site-specific integration system has been developed for mammalian cells. The invention system is based on the recombination of transfected sequences by FLP, a recombinase derived from Saccharomyces. In several cell lines, FLP has been shown to rapidly and precisely recombine copies of its specific target sequence. For example, a chromosomally integrated, silent .beta.-galactosidase reporter gene was activated for expression by FLP-mediated removal of intervening sequences to generate clones of marked cells. Alternatively, the reverse reaction can be used to target transfected DNA to specific chromosomal sites. These results demonstrate that FLP can be used, for example, to mosaically activate or inactivate transgenes for a variety of therapeutic purposes, as well as for analysis of vertebriate development.

Abstract: A process for preparing the sex pheromone of Lymantria dispar L. effectively which comprises the steps of reacting 1-bromodecane and propargyl alcohol tetrahydropyranylether in the presence of sodium hydroxide to give 1-tetrahydropyranyloxy-2-tridecyne, treating the tetrahydropyranyloxy-2-tridecyne with p-toluenesulfonic acid to give 2-tridecyn-1-ol, catalystic-hydrogenating the 2-tridecyn-1-ol in the presence of Lindlar catalyst to give (Z)-2-tridecen-1-ol, oxidizing the (Z)-2-tridecen-1-ol with peroxide to give (.+-.)-cis-2,3-epoxy-1-tridecanol, reacting the (.+-.

Abstract: The present invention relates to a DNA fragment of 3,563 base pairs containing a gene coding for tannase and derived from a microorganism belonging to the genus Aspergillus, with the following restriction enzyme map: ##STR1## B: Bam HI, H: Hind III, K: Kpn I, S: Sal I, X: Xba I; a DNA fragment containing a tannase gene coding for the amino acid sequence of (SEQ ID NO:4); a recombinant plasmid comprising the DNA fragment containing the tannase gene inserted into a plasmid vector; a process for producing tannase, comprising culturing a microorganism belonging to the genus Aspergillus capable of producing tannase in medium with the recombinant plasmid, and recovering tannase from the culture; and a promoter represented by the nucleotide sequence of (SEQ ID NO:1). Tannase can be efficiently produced according to the present invention.

Abstract: The present invention provides purified and isolated polynucleotide sequences encoding human plasma platelet-activating factor acetylhydrolase. Also provided are materials and methods for the recombinant production of platelet-activating factor acetylhydrolase products which are expected to be useful in regulating pathological inflammatory events.

Abstract: Conjugates of chlorophyll (Chl) and bacteriochlorophyll (Bchl) derivatives with amino acids, peptides and proteins are provided by the invention. The amino acid, peptide or protein residue is linked to the 17-propionic acid group of a Chl or Bchl residue directly or through a chain. The conjugates are for use as photosensitizers in photodynamic therapy and in diagnostics of tumors. Conjugation with cell-specific ligands, such as hormones, growth factors or tumor-specific antibodies, will target the Chl or Bchl moiety to the tumor site. Thus, conjugates with melanocyte stimulating hormones are suitable for photodynamic therapy of melanoma tumors.