Abstract: The current invention relates to methods of targeted genetic alteration in cells, preferably plant cells, as well as to plant cells and plants thus obtained using at least a fusion protein comprising a site-specific nuclease domain and a deaminase domain, or a construct encoding the same. The method also provides for a composition and a kit comprising a combination of a first fusion protein comprising a cytosine deaminase domain and a second fusion protein comprising an adenine deaminase domain, preferably for use in the method of the invention. The method provides for targeted alteration of a DNA duplex in plant cells with increased efficacy.

Abstract: The present invention provides a nitrilase mutant protein with increased thermal stability and its application in the synthesis of an anti-epileptic drug intermediate, wherein the mutant is obtained by mutating one or two of the amino acids at position 151, 223 and 250 of the amino acid sequence shown in SEQ ID No. 2. the thermal stability of the nitrilase mutant AcN-T151V/C223A/C250G was increased by up to 1.73 folds. The yield of the final product was up to 95% using the recombinant Escherichia coli containing the nitrilase mutant to hydrolyze 1M 1-cyanocyclohexylacetonitrile to produce 1-cyanocyclohexyl acetic acid at 35° C. And the yield of the final product was up to 97% when hydrolyzing 1.2M 1-cyanocyclohexylacetonitrile at 35° C. The final yield was up to 80% when using the nitrilase mutants obtained by the present invention to synthesize gabapentin.

Abstract: The present invention is directed to methods and compositions for adding tails of specific lengths to a substrate polynucleotide. The invention also contemplates methods and compositions for immobilization of tailed substrates to a solid support. The disclosure contemplates that the attenuator molecule is any biomolecule that associates with a tail sequence added to a substrate polynucleotide and controls the addition of a tail sequence to the 3? end of the substrate polynucleotide. The sequence that is added to the substrate polynucleotide is referred to herein as a tail sequence, or simply a tail, and the process of adding a nucleotide to a substrate polynucleotide is referred to herein as tailing.

Abstract: Compositions and methods are provided for genome modification of a target sequence in the genome of a cell, using a novel Cas endonuclease. The methods and compositions employ a guide polynucleotide/endonuclease system to provide an effective system for modifying or altering target sequences within the genome of a cell or organism. Also provided are novel effectors and endonuclease systems and elements comprising such systems, such as guide polynucleotide/endonuclease systems comprising an endonuclease. Compositions and methods are also provided for guide polynucleotide/endonuclease systems comprising at least one endonuclease, optionally covalently or non-covalently linked to, or assembled with, at least one additional protein subunit, and for compositions and methods for direct delivery of endonucleases as ribonucleotide proteins.

Abstract: The invention provides a method of modifying a targeted site of gram-positive bacterium of a double stranded DNA. The method includes contacting the double-stranded DNA with a complex of a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a given double stranded DNA and a nucleic acid base converting enzyme to convert, delete, or insert one or more nucleotides in the targeted site without cleaving at least one strand of the double stranded DNA in the targeted site, by introducing the nucleic acid encoding the complex into the gram-positive bacterium. The invention also provide a nucleic acid-modifying enzyme complex of a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a double stranded DNA of a gram-positive bacterium and a nucleic acid base converting enzyme bonded to each other, which complex is used for the method.

Abstract: The present invention is directed to methods and compositions for adding tails of specific lengths to a substrate polynucleotide. The invention also contemplates methods and compositions for immobilization of tailed substrates to a solid support. The disclosure contemplates that the attenuator molecule is any biomolecule that associates with a tail sequence added to a substrate polynucleotide and controls the addition of a tail sequence to the 3? end of the substrate polynucleotide. The sequence that is added to the substrate polynucleotide is referred to herein as a tail sequence, or simply a tail, and the process of adding a nucleotide to a substrate polynucleotide is referred to herein as tailing.

Abstract: Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. In some embodiments, fusion proteins of Cas9 and nucleic acid editing proteins or protein domains, e.g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e.g., fusion proteins of Cas9 and nucleic acid editing proteins or domains, are provided.

Abstract: Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. In some embodiments, fusion proteins of Cas9 and nucleic acid editing enzymes or enzyme domains, e.g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e.g., fusion proteins of Cas9 and nucleic acid editing enzymes or domains, are provided.

Abstract: The invention provides functional mutants of activation-induced cytidine deaminase (AID) protein that have increased activity as compared to a wild-type AID protein. The invention also provides nucleic acids encoding the functional AID mutants, and vectors and cells comprising the nucleic acids. The invention further provides methods of using the functional mutant AID proteins.

Abstract: The present invention relates to newly identified asparaginase polypeptide variants of SEQ ID NO: 3 and to polynucleotide sequences that encode such novel asparaginase variants. Furthermore the invention relates to the use of these novel asparaginase variants in industrial processes.

Abstract: The present invention provides a method of expressing at least one heterologous nucleic acid sequence in a cell, the method comprising introducing at least one heterologous nucleic acid sequence into a cell by infecting said cell with a recombinant negative-strand RNA virus vector comprising said at least one heterologous nucleic acid sequence, wherein the recombinant negative-strand RNA virus vector includes a viral genome coding for a mutated P protein, which leads to a loss of the viral genome replication ability without a loss of the viral transcription ability, and wherein said at least one heterologous nucleic acid sequence encodes a cellular reprogramming or programming factor or a therapeutic protein. In addition, the present invention provides a cell or a population of cells prepared in vitro by said method as well as a pharmaceutical composition comprising said cell or population of cells.

Abstract: The invention relates to nitrilases and to nucleic acids encoding the nitrilases. In addition methods of designing new nitrilases and method of use thereof are also provided. The nitrilases have increased activity and stability at increased pH and temperature.

Abstract: The present invention relates to methods of promoting the survival of cells by treating the cells with acid ceramidase. A kit for promoting ex vivo cell survival is also disclosed, as is a method of predicting in vitro fertilization outcome of a female subject.

Abstract: The subject of the invention is a modified gene of human activation-induced cytidine deaminase (AID), a method of modification of human AID gene, a composition showing AID activity, and a method of preparation of such composition. In particular, the invention concerns a modified gene of human AID for the production of an active enzyme in a bacterial system and use of the composition in the analyses of DNA/RNA amination/deamination and/or methylation/demethylatiori.

Abstract: The present invention provides polypeptides that bind to immunoglobulin G and methods for their use.

Abstract: The invention relates to a nitrilase having improved activity in the reaction of a nitrile to form the corresponding carboxylic acid, in particular with respect to reacting 2-methylglutaronitrile, 1-(cyanomethyl)cyclohexane-1-carbonitrile, and benzonitrile. The nitrilase according to the invention is related to nitrilase from acidovorax facilis.

Abstract: The present invention provides a site-directed mutated arginase and the preparation method thereof, and the use of said site-directed mutated arginase in preparing a medicament for treating an arginase-related disease. The present invention also provides a pegylated arginase and the preparation method thereof, and the use of said pegylated arginase in preparing a medicament for treating an arginase-related disease.

Abstract: The invention relates to nitrilases and to nucleic acids encoding the nitrilases. In addition methods of designing new nitrilases and method of use thereof are also provided. The nitrilases have increased activity and stability at increased pH and temperature.

Abstract: The invention provides histone deacetylase class II nucleic acids and polypeptides, methods and reagents for their use, and related compounds including small molecule libraries containing class II histone deacetylase inhibitors.

Abstract: The present invention relates to a polynucleotide that is active to an acetyl glutamate synthase and acetyl ornithinase which are associated with ornithine or arginine biosynthesis from Corynebacterium glutamicum. The present invention also relates to a polypeptide encoded by said polynucleotide, a recombinant vector comprising said polynucleotide, to a transformant obtained by introducing said recombinant vector to a host microorganism for producing L-ornithine or L-arginine, and transforming the recombinant vector, and to a method for producing L-ornithine or L-arginine by culturing said transformant. The activity of the transformant of the present invention to an acetyl glutamate synthase and acetyl ornithinase is increased as compared to an intrinsic activity, and thus L-ornithine or L-arginine can be produced, at a high yield rate, from the transformant of the present invention.

Abstract: This disclosure provides recombinant replication competent retroviral vectors having increased stability. The disclosure further relates compositions and uses of such vectors in the treatment of disease and disorders.

Abstract: Disclosed herein are polypeptides which exhibit asparaginase activity and little to no glutaminase activity. The polypeptides have sequences which correspond to W. succinogenes asparaginase, but have an amino acid residue other than proline at amino acid position 121. The polypeptides exhibit higher kinetic rates of asparaginase activity to glutaminase activity as compared to W. succinogenes asparaginase and Elspar®, an E. coli asparaginase.

Abstract: The present invention provides for novel compositions and methods for recycling or recovering ionic liquid used in IL pretreated cellulose and/or lignocellulosic biomass (LBM).

Abstract: The invention relates to nitrilases and to nucleic acids encoding the nitrilases. In addition methods of designing new nitrilases and method of use thereof are also provided. The nitrilases have increased activity and stability at increased pH and temperature.

Abstract: The present disclosure describes reproducible methods for the Agrobacterium-mediated production of stable, genetically transformed, fertile tarwi plants (Lupinus mutabilis Sweet) having seed-specific expression of human adenosine deaminase enzyme (hADA) or a functional variant thereof. The method involves slicing a tarwi seed embryo to produce an explant; infecting the explant in co-cultivation medium containing an agrobacterium having a polynucleotide sequence encoding hADA or variant; thereby generating a transformed explant; elongating a transformed shoot from the transformed explant; and regenerating a transformed tarwi plant from the elongated shoot. Seeds and plants so formed are also described herein. Further, methods for the recovery and purification of recombinant hADA, or functional variant from a transformed tarwi plant are described.

Abstract: What is provided is a method of treating a patient having a tumor comprising administering an effective amount of adenosine deaminase, preferably polyalkylene oxide conjugated, to a patient in need thereof.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: An object is to provide a novel method for improving an enzyme capable of deamidating a protein. A mutant protein deamidase is designed by the following steps: (A) identifying one or more amino acid in an amino acid sequence for a protein deamidase which corresponds to the amino acid at position 35, 38 to 43, 45, 46, 49, 79 to 84, 103 to 106, 117, 142, 143, 146, 166, or 185 in the amino acid sequence set forth in SEQ ID NO: 2; and (B) constructing a mutant amino acid sequence of the protein deamidase by substituting the one or more amino acid identified in step (A) with another amino acid or other amino acids or by deleting the one or more amino acid identified in step (A).

Abstract: The present invention provides a method for producing a cytosine deaminase (CD)-expressing, 5-fluorouracil (5-FU)-resistant microorganism which can grow in anaerobic tumor tissues, can express CD, and has a resistance to 5-FU at a concentration that is at least effective for antitumor activity. More specifically, the method is a method (A) comprising performing subculture or acclimation culture of a CD-expressing microorganism which can grow in anaerobic tumor tissues, in the presence of 5-fluorocytosine (5-FC), or a method (B) comprising (1) performing subculture or acclimation culture of a microorganism which can grow in anaerobic tumor tissues and does not express CD, in the presence of 5-FU to produce a 5-FU-resistant microorganism and (2) transforming the 5-FU-resistant microorganism by introducing a CD gene. The present invention also provides the CD-expressing, 5-FU-resistant microorganism and a pharmaceutical composition comprising the microorganism.

Abstract: This disclosure provides modified cytosine deaminases (CDs). The disclosure further relates to cells and vector expressing or comprising such modified CDs and methods of using such modified CDs in the treatment of disease and disorders.

Abstract: The invention relates to functionalized, telechelic polymers synthesized by enzymatic catalysis and methods, and the functionalization of polymers via Michael addition with a lipase catalyst, and the crosslinking of mono- or difunctional (telechelic) polymers made by enzymatic catalysis, such as by using multifunctional coupling agents and enzyme catalysts. Quantitative transesterification of vinyl methacrylate with poly(ethylene glycol), poly(isobutylene) and poly(dimethylsiloxane) was achieved using Candida antarctica lipase B. In addition, methacrylate-functionalized poly(ethylene glycol) monomethyl ether has been successfully coupled to aminoethoxy poly(ethylene glycol) monomethyl ether via Michael addition using Candida antarctica lipase B. Amine-functionalized poly(ethylene glycol)s have also been used for the preparation of poly(ethylene glycol)-based dendrimers and gels through Michael addition of the polymer onto triacryloyl hexahydro-triazine using the same enzyme.

Abstract: The present disclosure relates to a deacetylation hydrolase of a hyaluronic acid a hyaluronic acid deacetylated by same and a derivative thereof. The deacetylated hyaluronic acid and the derivative thereof have the following characteristic: a delayed initial decomposition rate on a living body; minimized decrease of molecular weight and viscosity; accelerated gelation due to a lower gelation temperature than the gelation temperature for a non-deacetylated hyaluronic acid; and an hMSC survival rate that is hardly affected by increased concentration of the deacetylated hyaluronic acid and the derivative thereof in a culture medium. As a result, the deacetylated hyaluronic acid and the derivative thereof can be useful as a bioingredient such a delivery system for a cell, gene, drug, and the like, or a support for tissue engineering, etc.

Abstract: The object of the present invention is to provide a fluorescent substrate for detecting the enzymatic activity of a nitrile-related enzyme. The present invention provides a compound represented by formula (I) and a fluorescent substrate for detecting the enzymatic activity of a nitrile-related enzyme, which comprises the compound.

Abstract: The present invention provides an isolated and substantially purified recombinant human arginase having sufficiently high enzymatic activity and stability to maintain Adequate Arginine Depletion in a patient. The present invention also provides a pharmaceutical composition comprising the modified invention enzyme and method for treatment of diseases using the pharmaceutical composition.

Abstract: The present invention relates to an arginine deiminase mutant with partial lysine-deficient and preparation and application thereof. The arginine deiminase mutant of the present invention has enzymatic activity of degrading arginine into citruline; compared with the arginine deiminase with the amino acid sequence of SEQ ID NO: 1, the amino acid sequence comprises one or more of K9N, T, K59Q, K66R, A, K93E, A, Q, K111R, A, K119Q, L, M, K121Q, I, K122E, L, K126E, S, R, K178I, E, D, K196I, R, K209G, T, D, K243E, V, R, K249D, Q, K263N, Q, K279Y, T, K293R, H, E, K325V, I, K380T, R, E, and K406E, D, S substitutions. Compared with PEG modified natural derived arginine deiminase, the PEG modified arginine deiminase mutant of the present invention retain better bioactivity; and because the quantity of lysine in arginine deiminase is reduced, the PEG modified products are more uniform and can be applied to clinical treatment of hepatoma, melanoma and the like.

Abstract: The present invention relates to novel nucleic acid molecules for producing target polypeptides in plant cells. More specifically, the novel nucleic acid molecules comprise a minireplicon derived from a Closteroviridae virus and heterologous polynucleotides encoding the target polypeptides. Also provided are compositions comprising the target polypeptides and uses thereof.

Abstract: The present invention is intended to provide a quality improving agent useful for improving the quality of steamed buns, and a quality improving agent containing glucoamylase. The quality improving agent preferably has low protease activity.

Abstract: The present invention relates to newly identified asparaginase polypeptide variants of SEQ ID NO: 3 and to polynucleotide sequences that encode such novel asparaginase variants. Furthermore the invention relates to the use of these novel asparaginase variants in industrial processes.

Abstract: The present disclosure relates to polypeptides having nitrilase activity, polynucleotides encoding the polypeptides, and methods of using the polypeptides.

Abstract: This invention is related to the field of the prevention and treatment of kidney disease. The treatment of kidney disease may be tailored depending upon the need for, or expectation of, renal recovery. For example, renal recovery can be determined by monitoring urine biomarkers related to the development of chronic kidney disease. For example, a normalized time course of approximately fourteen Days measuring urinary proteins can be used to establish the risk of recovery versus non-recovery in patient's having suffered an acute kidney injury. Alternatively, the invention describes signature protein expression profiles to establish the probability of renal to recovery and/or renal non-recovery.

Abstract: Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods of at least partially degrading, cleaving, or removing polysaccharides, lignocellulose, cellulose, hemicellulose, lignin, starch, chitin, polyhydroxybutyrate, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius.

Abstract: The invention relates to nitrilases and to nucleic acids encoding the nitrilases. In addition methods of designing new nitrilases and method of use thereof are also provided. The nitrilases have increased activity and stability at increased pH and temperature.

Abstract: The invention relates to a nitrilase having improved activity in the reaction of a nitrile to form the corresponding carboxylic acid, in particular with respect to reacting 2-methylglutaronitrile, 1-(cyanomethyl)cyclohexane-1-carbonitrile, and benzonitrile. The nitrilase according to the invention is related to nitrilase from acidovorax facilis.

Abstract: Compositions and methods are provided for discrimination between cytosine and modifications thereof using cytidine deaminases and/or oxygenases. Variants of wild type cytidine deaminases are described which show reduced bias with respect to adjacent nucleotides upstream of the cytosine. The methods provide a rapid and convenient use of enzymes to obtain methylomes.

Abstract: This invention relates to a novel, bacterial GTP Cyclohydrolase Type IB enzyme, and the crystal structure thereof.

Abstract: The present invention provides diagnostic methods for determining the risk of developing an autism spectrum disorder (ASD) in a fetus or child by detecting in a biological sample from the mother antibodies that bind to one or more biomarkers selected from the group consisting of lactate dehydrogenase (LDH), guanine deaminase (GDA), collapsin response mediator protein 1 (CRMP1), stress-induced phosphoprotein 1 (STIP1), alpha subunit of the barbed-end actin binding protein Cap Z (CAPZA2), Y Box Binding Protein 1 (YBX1), eukaryotic translation and elongation factor 1A1 (EEF1A1), microtubule-associated protein Tau (MAPT), dihydropyrimidinase-like protein 2 (DPYSL2), dynamin 1-like protein (DNM1L), radixin (RDX), moesin (MSN), and ezrin (EZR).

Abstract: Mono-pegylated arginase conjugate and method producing thereof. The mono-pegylated arginase is homogeneous in molecular weight and shows therapeutic effect for treating cancers and viral infections. The method of producing such arginase conjugate has a main step of genetically modifying the gene encoding an arginase so that the PEG moiety can attach to the enzyme at a predetermined, specific intended site. This is achieved by removing the PEG attaching amino acid residues at undesirable sites while keeping (or adding, if necessary) the one at the desirable site of the enzyme. Two exemplary mono-pegylated arginase conjugates so produced are human arginase I (HAI) where a polyethylene glycol (PEG) moiety is site-specific covalently bonded to Cys45 of the enzyme and Bacillus caldovelox arginase (BCA) where a polyethylene glycol (PEG) moiety is site-specific covalently bonded to Cys161 of the enzyme.

Abstract: This disclosure concerns compositions and methods for targeting peptides, polypeptides, and proteins to plastids of plastid-containing cells. In some embodiments, the disclosure concerns chloroplast transit peptides that may direct a polypeptide to a plastid, and nucleic acid molecules encoding the same. In some embodiments, the disclosure concerns methods for producing a transgenic plant material (e.g., a transgenic plant) comprising a chloroplast transit peptide, as well as plant materials produced by such methods, and plant commodity products produced therefrom.

Abstract: The invention provides nitrilases and methods for making and using them, and in one aspect, provides methods for producing enantiomerically pure ?-substituted carboxylic acids, such as, for example, ?-amino acids and ?-hydroxy acids. In one aspect, methods of the invention combine an aldehyde or ketone with a cyanide and ammonia or an ammonium salt or an amine, in the presence of a nitrilase or a polypeptide having nitrilase activity, to stereoselectively hydrolyze the amino nitrile or cyanohydrin intermediate under conditions sufficient to produce the carboxylic acid.

Abstract: The present invention relates to an agent for use as a modulator of apoptosis-factor-associated cell death, apoptosis, cell survival, migration and/or proliferation, a method of diagnosing or monitoring apoptosis-factor-associated conditions or disorders as well as a method of identifying a modulator of apoptosis-factor-associated cell death, apoptosis, cell survival, migration and/or proliferation. Preferably, the invention relates to TRAIL-induced cell death and/or TRAIL-induced apoptosis. Preferably the agents are used to stimulate and/or enable TRAIL-induced cell death or to inhibit TRAIL-induced cell death. The preferable use of the diagnostic tools is to diagnose sensitivity or resistance to TRAIL-induced cell death or induction of sensitivity or resistance to TRAIL-induced cell death by an agent.