Abstract: By using engineered sequence specific DNA nuclease (“SSDN”), the composition, reagent kit and method of the present invention can cut and release a DNA sequence of interest 1×104-1×107-base pairs long from a source DNA as large as the whole genome. The SSDN further includes an affinity tag or is bound to a solid support that facilitates the isolation of the DNA sequence of interest. The SSDN can include a RecA and Ref combination, a transcription activator like effector nuclease, or a sequence specific chemical nuclease. When applied to genomic sequencing, specific region(s) of interest in the genome can be cut and isolated. Because the irrelevant part of the genome is removed from the sequencing reaction, the speed, cost, and accuracy of genomic sequencing can be improved.

Abstract: The present invention consists of methods for joining DNA molecules (parts) together to form larger DNA molecules (assemblies) of specified sequence and organization. The invention exhibits three necessary characteristics. Firstly, the invention enables 2 or more parts to be joined in a single reaction. Secondly, the seam between joined parts is scarless, producing no residual sequence dependencies like restriction enzyme recognition sites. Thirdly, parts are modular and can easily be reused in novel assemblies without modification. Prior technologies have exhibited no more than two of the three necessary characteristics, limiting their utility in synthesizing and editing DNA molecules of arbitrary sequence.

Abstract: Polypeptide and recombinant protein having esterase activity which exhibit the amino acid sequence SEQ. ID. No. 1 and the use thereof.

Abstract: The present invention provides AP that has a high specific activity and preferably has superior reactivity with respect to various luminescent substrates generally used for high-sensitivity immunoassay analysis. Further preferably, the present invention provides AP having a thermal stability higher than that of CIAP. The alkaline phosphatase of the present invention is derived from the genus Shewanella and has the characteristics below: (A) molecular weight: about 104,000; (B) optimum reaction pH: about 9.5; (C) stable pH range: 5.5 to 10.4; (D) thermal stability: 65° C.; and (E) specific activity: 5,000 U/mg or more.

Abstract: The present invention relates to variants (mutants) of parent Termamyl-like alpha-amylases, which variant has alpha-amylase activity and exhibits altered stability, in particular at high temperatures and/or at low pH relative, and/or low Ca2+ to the parent alpha-amylase.

Abstract: The present invention relates to novel polynucleotide sequences comprising genes that encode novel lipolytic enzymes, as well as functional equivalents of the gene or the amino acid sequences with high homology thereto. The invention also relates to methods of using these lipolytic enzymes in industrial processes, for example in the dairy or baking industry.

Abstract: The present invention provides fungal xylanase and/or xylosidase enzymes suitable for use in saccharification reactions. The present invention provides xylanase and xylosidase enzymes suitable for use in saccharification reactions. The present application further provides genetically modified fungal organisms that produce xylanase(s) and/or xylosidase(s), as well as enzyme mixtures exhibiting enhanced hydrolysis of cellulosic material to fermentable sugars, enzyme mixtures produced by the genetically modified fungal organisms, and methods for producing fermentable sugars from cellulose using such enzyme mixtures. In some embodiments, the xylanase and xylosidase enzyme(s) are M. thermophila enzymes.

Abstract: The present invention provides, among other things, methods and compositions for production of recombinant I2S protein with improved potency and activity using cells co-express I2S and FGE protein. In some embodiments, cells according to the present invention are engineered to simultaneously over-express recombinant I2S and FGE proteins. Cells according to the invention are adaptable to various cell culture conditions. In some embodiments, cells of the present invention adaptable to a large-scale suspension serum-free culture.

Abstract: The present invention provides, among other things, methods and compositions for large-scale production of recombinant I2S protein using suspension culture of mammalian cells in serum-free medium. In particular, the present invention uses mammalian cells co-express a recombinant I2S protein and a formylglycine generating enzyme (FGE).

Abstract: The present invention provides, among other things, improved methods for purifying I2S protein produced recombinantly for enzyme replacement therapy. The present invention is, in part, based on the surprising discovery that recombinant I2S protein can be purified from unprocessed biological materials, such as, I2S-containing cell culture medium, using a process involving as few as four chromatography columns.

Abstract: The present invention provides polypeptide derivatives of IGFBP-3 that are resistant to proteolytic cleavage. These IGFBP-3 derivatives are useful in a variety of therapeutic and diagnostic applications. Also provided are pharmaceutical compositions and kits comprising such IGFBP-3 derivatives and methods for using these derivatives for the treatment of a variety of disorders.

Abstract: Antibodies binding to sites on the alpha-subunit of the (Na++K+)-ATPase increase cardiac contraction of both ventricular myocytes and mouse heart. In particular, antibodies binding to the RSATEEEPPNDD (SEQ ID NO: 1) or DVEDSYGQQWTYEQR (SEQ ID NO: 2) peptides (or isoforms/derivatives thereof) of the alpha-subunit of the (Na++K+)-ATPase, have been found to be highly inotropic. Both the antibodies and the peptides are important for the treatment of human heart failure and other contractile disorders.

Abstract: A class of anionic oligomers and polymers that function for inhibition of nucleases, particularly RNase. Specific inhibitors include mixtures of oligomers of vinyl sulfate. Methods for inhibition or inactivation of one or more nucleases in vitro which comprises the step of contacting the one or more nucleases in a biological medium with one or more of the anionic oligomeric or polymeric inhibitors of this invention. Kits for carrying out a biological procedure, biological reaction and/or a biological assay containing one or more inhibitors of this invention. The use of oligomers and/or polymers of this invention as additives in buffers or reagents. The inhibitors of the invention can also be attached to surfaces to provide for removal of nucleases from media, solutions or other liquids in contact with the solid.

Abstract: A heat labile alkaline phosphatase enzyme and methods of using the same and kits including the same are disclosed. Specifically, a nucleotide sequence of, peptide sequence of, methods of using, and kits comprising, a heat labile alkaline phosphatase isolated from Colwellia psychrerythraea are provided. Methods of over-expression and purification of the recombinant alkaline phosphatase and mutants thereof are also disclosed. Methods of over-expressing and purifying commercially useful quantities of active recombinant heat labile alkaline phosphatase fusion enzymes from C. psychrerythraea, wherein the fusion enzymes comprise one or more heterologous leader sequences are disclosed. The disclosed C. psychrerythraea heat labile alkaline phosphatase has properties similar to shrimp alkaline phosphatase and can be substituted for shrimp alkaline phosphatase in assays involving the same.

Abstract: A method includes combining a polynucleotide and an amplification reagent mixture to form a reaction mixture, wherein the reaction mixture comprises reversibly bound divalent ions in solution, and adjusting the pH of the reaction mixture to release the reversibly bound divalent ions, thereby initiating amplification of the polynucleotide.

Abstract: Disclosed is a method for production of recombinant human iduronate 2-sulfatase (rhI2S) in a large scale, with a high purity, and mannose 6-phosphate residues. The method comprises the steps of (a) culturing rhI2S-producing mammalian cells in a serum-free medium, (b) collecting culture supernatant, (c) subjecting the culture supernatant to cation-exchange column chromatography, (d) to dye affinity column chromatography, (e) to anion-exchange column chromatography, and (f) to a column chromatography employing as solid phase a material having affinity for phosphate group, and (g) to gel filtration column chromatography, in the order.

Abstract: A LAGLIDADG homing endonuclease variant, having mutations in two separate subdomains, each binding a distinct part of a modified DNA target half-site, said LAGLIDADG homing endonuclease variant being able to cleave a chimeric DNA target sequence comprising the nucleotides bound by each subdomain. Use of said herodimeric meganuclease and derived products for genetic engineering, genome therapy and antiviral therapy.

Abstract: The present invention provides methods, compositions, and kits for the treatment of neurocutaneous syndromes, such as neurofibromatosis type I; disorders associated with overactivation of FGFR3, such as achondroplasia; bone or cartilage disorders; or vascular smooth muscle disorders; or for the elongation of bone. In some embodiments, the present invention provides polypeptides having an alkaline phosphatase peptide fused to an Fc domain of an immunoglobulin or a natriuretic peptide fused to an Fc domain of an immunoglobulin. Such polypeptides can be administered to subjects, e.g., subcutaneously, to treat a neurocutaneous syndrome, a disorder associated with overactivation of FGFR3, a bone or cartilage disorder, or a vascular smooth muscle disorder, or to elongate bone.

Abstract: The invention provides methods, compositions, and kits for removal of biofilms from surfaces. The methods described herein comprise simultaneous or sequential application of a perhydrolase enzyme and a mixture of other enzymes, such as proteases, glucanases, esterases, mannanases, phospholipases, cellulases, and/or amylases, to a biofilm on a surface, to effect removal of the biofilm.

Abstract: Butyrylcholinesterase (BChE) polypeptide variants of the presently-disclosed subject matter have enhanced catalytic efficiency for (?)-cocaine, as compared to wild-type BChE. Pharmaceutical compositions of the presently-disclosed subject matter include a BChE polypeptide variant having an enhanced catalytic efficiency for (?)-cocaine. A method of the presently-disclosed subject matter for treating a cocaine-induced condition includes administering to an individual an effective amount of a BChE polypeptide variant, as disclosed herein, to lower blood cocaine concentration.

Abstract: The invention provides methods for enriching methyl-CpG sequences from a DNA sample. The method makes use of conversion of cytosine residues to uracil under conditions in which methyl-cytosine residues are preserved. Additional methods of the invention enable to preservation of the context of me-CpG dinucleotides. The invention also provides a recombinant, full length and substantially pure McrA protein (rMcrA) for binding and isolation of DNA fragments containing the sequence 5?-CMeCpGG-3?. Methods for making and using the rMcrA protein, and derivatives thereof are provided.

Abstract: The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).

Abstract: The invention relates to the field of medicine and in particular to the use of alkaline phosphatase in the treatment of renal diseases, such as reduced renal function. The present invention also relates to the field of pharmacy and in particular to the pharmaceutical use of alkaline phosphatase. The present invention provides an alternative treatment to improve a situation in which the renal function is reduced by using alkaline phosphatase.

Abstract: A two-stage heat treatment process is provided to improve the processability of recombinant microbial biomass comprising an enzyme having perhydrolytic activity. More specifically, a process is provided to treat the recombinant microbial biomass comprising a Thermotoga sp. acetyl xylan esterase having perhydrolytic activity such that microfiltration may be used to partially-purify and/or concentrate protein preparations. The acetyl xylan esterase may be used to produce peroxycarboxylic acids suitable for use in a variety of applications such as cleaning, disinfecting, sanitizing, bleaching, wood pulp processing, and paper pulp processing applications.

Abstract: Ethylenically unsaturated glycosides of the formula I: wherein n, A, X, R3 and R4 have the meanings given in the description are produced by reacting an ethylenically unsaturated alcohol of formula II: with a saccharide of formula III: in the presence of a glucosidase at a molar ratio of the alcohol to the saccharide of from 2:1 to 30:1 in the presence of a solvent mixture of water and 1,4-dioxane at a weight ratio of 0.1:1 to 9:1 and a weight ratio of solvent mixture to saccharide of from 3:1 to 30:1.

Abstract: The invention relates to agents containing esterases, and to the use thereof for dressing fibres, in particular, artificial fibres, washing and cleaning agents comprising esterases and corresponding washing and cleaning methods, in addition to additional technical areas of application. The invention also relates to the use of esterases for protecting against or reducing and/or preventing pilling, preferably in textiles, particularly plastic fibres, more preferably polyester fibres, in addition to the use of esterases for separating the plastics, in particular, polyester compounds. The invention further relates to novel esterases and to sufficiently related proteins and to derivatives thereof, agents containing them and to the use thereof.

Abstract: The present invention relates to polypeptides having phytase activity. These polypeptides have an amino acid sequence which has at least 70% identity to either of three phytases derived from the bacterium Buttiauxella, and which comprises at least one of the following amino acids at the position indicated: 119N, 120L, and/or 121E. These phytases have an improved specific activity. Additional specific amino acid substitutions are also disclosed which characterize and distinguish additional phytases of the invention having improved properties such as temperature and/or pH stability, pH activity profile, temperature activity profile, substrate profile, improved performance in animal feed in vitro or in vivo. The invention also relates to isolated polynucleotides encoding the polypeptides, nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods for producing and using the polypeptides.

Abstract: A DNA library, and a preparing method thereof, a method of determining DNA sequence information, an apparatus and a kit for detecting SNPs, and a method for genotyping may be provided. The method for preparing the DNA library may comprise the steps of: digesting a genomic DNA sample using a restriction endonuclease to obtain a digested product, wherein the restriction endonuclease comprises at least one selected from the group consisting of Mbo II and Tsp 45I; separating the digested product to obtain DNA fragments having a length of 100 bp to 1,000 bp; end-repairing the DNA fragments to obtain an end-repaired DNA fragments; adding a base A to the end of the end-repaired DNA fragments to obtain DNA fragments having a terminal base A; and ligating the DNA fragments having the terminal base A with sequencing adaptors to obtain the DNA library.

Abstract: Provided herein are methods and compositions for treating a subject suffering from a deficiency in arylsulfatase A in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an arylsulfatase A.

Abstract: The invention features methods for producing isoprene from cultured cells. The invention also provides compositions that include these cultured cells. The invention provides isoprene compositions, such as compositions with increased amount of isoprene or increased purity. Additionally, the invention provides methods of producing isoprene by culturing cells under conditions suitable for isoprene production while maintaining cell viability and/or metabolic activity.

Abstract: The present invention relates to novel enzymes and the uses thereof. The invention also relates to methods of producing such enzymes, coding nucleic acid molecules, recombinant cells and methods of transforming biomass from such materials. The invention is particularly suited to degrade biomass and/or to improve biomass degradation, and to produce bioenergy products or recombinant proteins. This invention also relates to various applications of the enzymes in the field of paper industry, textile industry as well as in the chemical and medical fields.

Abstract: The present invention relates to modified filamentous fungal organisms having improved activity profiles with respect to the conversion of complex carbohydrates into simple sugars from cellulosic materials, including fungal organisms belonging to a genus selected from the group consisting of: Chrysosporium, Thielavia, Talaromyces, Thermomyces, Thermoascus, Neurospora, Aureobasidium, Filibasidium, Piromyces, Corynascus, Cryplococcus, Acremonium, Tolypocladium, Scytalidium, Schizophyllum, Sporotrichum, Penicillium, Gibberella, Myceliophthora, Mucor, Aspergillus, Fusarium, Humicola, Trichoderma, and Talaromyces, plus anamorphs and teleomorphs thereof.

Abstract: A computer implemented method for generating nucleotide sequences containing candidate homing endonuclease genes (HEGs). A search is performed in a database stored on a storage medium of nucleotide sequences for amino acid sequences having a subsequence having a homology level with the translation of a subsequence of one or more predetermined HEGs. For each amino acid sequence generated by the search, one or more nucleotide sequences are retrieved encoding the amino acid sequence. The results of this search used in a second search of a database stored on a storage medium to generate the HEG containing sequences.

Abstract: The invention relates to a synthetic phytase with elevated thermostability, elevated stability to acids at pH 2, elevated stability to pepsin and with a broadened active pH range, and to an isolated nucleic acid sequence coding for a synthetic phytase and to the use of the phytase in an animal feed for reducing the phosphate content in the slurry and to animal feed additives and animal feeds comprising the synthetic phytase.

Abstract: A method is described for improving the nutritional value of a foodstuff comprising a source of myo-inositol hexakisphosphate by feeding the foodstuff in combination with a phytase expressed in yeast. The method comprises the step of feeding the animal the foodstuff in combination with a phytase expressed in yeast wherein the phytase can be selected from the group consisting of AppA1, AppA2 and a site-directed mutant of AppA. The invention also enables reduction of the feed to weight gain ratio and an increase bone mass and mineral content of an animal. A foodstuff and a feed additive comprising AppA2 or a site-directed mutant of AppA are also described.

Abstract: The present invention relates to processes for the preparation of 5-methyl-3-nitromethyl-hexanoic acid ester and its salts. Also disclosed are processes for the preparation of 5-methyl-3-nitromethyl-hexanoic acid salt and a process for the preparation of 3-(aminomethyl)-5-methylhexanoic acid. (S)-5-Methyl-3-nitromethyl-hexanoic acid or (R)-5-methyl-3-nitromethyl-hexanoic acid in enantioenriched form or enantiopure form as well as salts thereof, (S)-5-methyl-3-nitromethyl-hexanoic acid ester or (R)-5-methyl-3-nitromethyl-hexanoic acid ester in enantioenriched form or enantiopure form and a compound, namely Formula (XIII), in racemic form, enantioenriched form or enantiopure form are also disclosed.

Abstract: This invention provides an alkaline phosphatase or an active derivative of placental alkaline phosphatase to prevent or reduce one, two, or all three of interrelated metabolic disorders, i.e. insulin resistance, hyperinsulinemia, and dyslipidemia. Alkaline phosphatase or an active derivative of placental alkaline phosphatase is suitable to prevent or reduce diseases and disorders relating to insulin resistance, hyperinsulinemia, and dyslipidemia including type 2 diabetes, epithelial cancers, myopia, acne, acanthosis nigricans, polycystic ovary syndrome, cutaneous papillomas, hypertension, renal dysfunction, microvascular and macrovascular diseases, atherosclerosis, nonalcoholic fat liver, dyslipidemia, sleep-disordered breathing, atherosclerosis, coronary artery (heart) disease, cardiovascular disease, heart failure, accelerated aging, stroke, and neurological diseases such as Alzheimer.

Abstract: Provided are a bladder cancer diagnosis composition containing APE1/Ref-1, a bladder cancer diagnostic kit containing an antibody which specifically binds to the APE1/Ref-1, and a method of measuring APE1/Ref-1 concentration in biological samples through an antigen-antibody binding reaction using the antibody which specifically binds to the APE1/Ref-1. According to the invention, the APE1/Ref-1 protein concentration in serum of bladder cancer patients is significantly higher than in healthy subjects, and more particularly, it significantly increases in the serum of patients with stage 2 or later bladder cancer.

Abstract: This invention relates generally to enzymes, polynucleotides encoding the enzymes, the use of such polynucleotides and polypeptides and more specifically to enzymes having isomerase activity, e.g., racemase activity, e.g., amino acid racemase activity, alanine racemase activity, and/or epimerase activity, and/or catalyze the re-arrangement of atoms within a molecule, catalyze the conversion of one isomer into another, catalyze the conversion of an optically active substrate into a raceme, which is optically inactive, catalyze the interconversion of substrate enantiomers, catalyze the stereochemical inversion around the asymmetric carbon atom in a substrate having only one center of asymmetry, catalyze the stereochemical inversion of the configuration around an asymmetric carbon atom in a substrate having more than one asymmetric center, and/or catalyze the racemization of amino acids.

Abstract: The invention concerns compositions and methods combining at least two phytases for hydrolyzing phytic acid (myo-inositol hexakis phosphate) into inorganic monophosphates, into myo-inositols with lower degree of phosphorylation and into free myo-inositol. Said compositions and methods are of particular interest for animal feeding.

Abstract: The invention provides variant phytase enzymes having increased thermal stability relative to their counterpart parent enzymes. The modifications to the enzymes include both single substitutions and various combinations of substitutions that provide improved stability and activity. The invention further provides nucleic acids encoding the variant phytase enzymes, host cells and vectors containing and expressing them, as well as feed compositions useful for providing improved nutrition, particularly with respect to the bioavailability of dietary phosphate, calcium, iron and zinc, among others.

Abstract: The present invention relates to a method for genetically modifying a filamentous fungus host for improved protein production. The method comprises that a filamentous fungus host is genetically modified to be deficient or to overexpress specific genes. The invention relates also to the modified hosts. Furthermore, the invention relates to a method for improved production or for producing an improved composition of proteins, such as cellulases, hemicellulases, other proteins involved in the degradation of lignocellulosic material, or other proteins, in a filamentous fungus host.

Abstract: The present invention relates to a method of producing a heterologous protein or polypeptide having phytase activity in a yeast system. The invention also provides proteins having phytase activity which have increased thermostability. Yeast strains which produce a heterologous phytase and the vectors used to produce the phytase are also provided.

Abstract: The present invention pertains to a process for production of recombinant arylsulfatase A in a cell culture system, the process comprising culturing a mammalian cell capable of producing rASA in liquid medium in a system comprising one or more bio-reactors; and concentrating, purifying and formulating the rASA by a purification process comprising one or more steps of chromatography. Other aspects of the invention provides a pharmaceutical composition comprising rASA, which is efficiently endocytosed via the mannose-6-phosphate receptor pathway in vivo as well as a rhASA a medicament and use of a rhASA for the manufacture of a medicament for reducing the galactosyl sulphatide levels within target cells in the peripheral nervous system and/or within the central nervous system in a subject.

Abstract: Modified DNase polypeptides and methods of their use are provided.

Abstract: The invention is directed to treatment of delayed-type hypersensitivity reactions associated with changes of qualitative and/or quantitative composition of blood extracellular DNA and treatment of systemic DNA mutation diseases accompanied with development of somatic mosaicism and elevation of blood extracellular DNA. The inventive method comprises introducing a DNASE enzyme into the systemic blood circulation of a patient in doses and regimens which are sufficient to decrease average molecular weight of circulating extracellular blood DNA in the blood of said patient.

Abstract: Disclosed is a modified iduronate-2-sulfatase (IDS) gene constructed by inserting the nucleotide of SEQ ID NO: 2 into a wild-type IDS gene. In addition to being negatively charged, the improved IDS enzyme encoded by the modified gene exhibits a sufficient retention time in blood to target the bone, so that it is more effective for treating Hunter syndrome.

Abstract: An I-CreI variant, wherein one of the two I-CreI monomers has at least two substitutions, one in each of the two functional subdomains of the LAGLIDADG core domain situated respectively from positions 26 to 40 and 44 to 77 of I-CreI, said variant being able to cleave a DNA target sequence from the mouse ROSA26 locus. Use of said variant and derived products for the engineering of transgenic mice and recombinant mouse cell lines expressing an heterologous protein of interest.

Abstract: This invention relates to altered forms of members of the RNase A superfamily. An RNase A can be modified to be cytotoxic by altering its amino acid sequence so that it is not bound easily by the ribonuclease inhibitor while still retaining catalytic properties. While earlier work had identified some modifications to RNase A that would result in cytotoxicity, the use of the FADE algorithm for molecular interaction analysis has led to several other locations that were candidates for modification. Some of those modifications did result in RNase A variants with increase cytotoxicity.

Abstract: Rationally-designed LAGLIDADG meganucleases and methods of making such meganucleases are provided. In addition, methods are provided for using the meganucleases to generate recombinant cells and organisms having a desired DNA sequence inserted into a limited number of loci within the genome, as well as methods of gene therapy, for treatment of pathogenic infections, and for in vitro applications in diagnostics and research.