Abstract: The invention relates to a method for synthesizing a peptide by enzymatically preparing an ester or thioester from (i) an N-terminal protected amino acid or an N-terminal protected peptide where either can have a protected C-terminal ester group and (ii) an alcohol represented by the formula HO—CX2—Z or a thiol represented by the formula HS—CX2—Z, each X independently representing a halogen atom or a hydrogen atom; and Z represents an electron withdrawing group comprising at least one sp3-hybridized carbon comprising at least two substituents comprising a heteroatom directly attached to the at least one sp3-hybridized carbon or at least one sp2-hybridized carbon comprising one or two substituents comprising a heteroatom directly attached to the at least one sp2-hybridized carbon, and enzymatically coupling the prepared ester or thioester with an optionally C-terminal protected amino acid or with an optionally C-terminal protected peptide in a medium comprising 2 wt. % water or less.

Abstract: A use of a signal peptide for producing a recombinant polypeptide of interest in an expression system, the signal peptide includes at least 12 amino acids of formula (I): (X1)iX2X3X4SX5X6X7, wherein: X1 is a peptide containing from 3 to 6 amino acids, i equal to 0 or 1, X2 is a peptide containing from 3 to 9 hydrophobic amino acids, X3 is a peptide containing from 3 to 5 amino acids, the peptide including at least 3 contiguous or non-contiguous leucines X4 is a peptide containing from 2 to 5 amino acids chosen from Ala, Thr, Ser, Gln, Ile, Met, X5 is Ala or Val, X6 is Gln, Asn or His, X7 is Ala or Cys, provided that when the signal peptide originates from a natural precursor of a specific protein, the polypeptide of interest is different from the protein.

Abstract: The invention relates to a dried enzymically hydrolyzed plant material, which is capable of immunmodulation, immunoregulation, reducing inflammation and reducing tumor progression. The material can be used in the manufacture of pharmaceutical compositions, dietary supplements, dietary foods for medical purposes, nutraceuticals, in feed materials, and in supplementary feeds. The process for producing the material is also described, as are its use in the modulation, and regulation of immune reactions, reducing inflammation and/or tumor progression. The material can also be used in treatment of several inflammatory disorders.

Abstract: A non-therapeutic method of accumulating a polymeric or high molecular weight molecular product within a bacterial microcompartment in bacterial cytoplasm, which method employs a recombinant bacteria which is transformed to express a microcompartment containing an enzyme capable of converting a low molecular weight substrate into a polymeric or high molecular weight product, the method comprising the steps of: incubating the recombinant bacteria with the low-molecular weight substrate, or a precursor of the low molecular weight substrate which is capable of being metabolised to the substrate within the recombinant bacteria, such that the substrate or precursor is taken up by the bacteria, wherein the substrate enters the microcompartment and the enzyme within the microcompartment converts the substrate to a polymeric or high molecular weight molecular product, and wherein the polymeric or high molecular weight molecular product is accumulated within the microcompartment due to its size.

Abstract: The invention provides a polypeptide containing at least one IgG Fc region region, said polypeptide having a higher anti-inflammatory activity and a lower cytotoxic activity as compared to an unpurified antibody and methods of production of such polypeptide.

Abstract: The present invention provides a method for labeling or detecting a protein with certain glycosyl groups. The methods are particularly useful for detecting cancer cells comprising the detected glycosyl groups. The present invention further provides labeling agents and detection agents, labeled proteins and mixtures, and kits and arrays thereof.

Abstract: Method for incorporating a lysine derivative (particularly an N?-benzyloxycarbonyl-lysine (Z-Lys) derivative) having useful functional group such as heavy atom, selenium, reactive functional group, fluorescent group or crosslinker, which is suitable as a non-natural amino acid, into a desired protein in a site-specific manner. A mutant pyrrolysyl-tRNA synthetase has substitution of at least one amino acid residue selected from tyrosine residue at position 306, leucine residue at position 309 and cysteine residue at position 348 each constituting a pyrrolysine-binding site in the amino acid sequence for pyrrolysyl-tRNA synthetase of SEQ ID NO:2. The substitution of the amino acid residue is: of tyrosine residue at position 306 by glycine or alanine residue, of leucine residue at position 309 by glycine or alanine residue, and/or of a cysteine residue at position 348 by valine, serine or alanine residue.

Abstract: A fungal strain Beauveria species bearing accession number MTCC 5184 is disclosed. The process for the preparation of an enzyme mix including at least one enzyme selected from, but not limited to protease, carbohydrase, and lipase from the disclosed Beauveria species and uses of the enzyme mix in various areas also disclosed.

Abstract: Methods are provided for a one step synthesis of polypeptide polymers or co-polymers. The polymers or co-polymers can be linear or branched. In the methods of the invention, the coding sequence for the polypeptide(s) to be polymerized is altered by introducing one or more codons for an nonnatural amino acid, which coding sequence is then utilized to produce the cognate polypeptide. The nonnatural amino acids are selected to be reactive with each other in a bioorthogonal reaction, and are combined in a conjugation reaction with the desired components of the polymer.

Abstract: An active small-molecule donkey-hide gelatin mixture and a preparation method and application thereof. The active small-molecule donkey-hide gelatin mixture is prepared by using the compound protease comprising the proline protease to perform the enzymatic hydrolysis on the donkey-hide gelatin juice to which no auxiliary material is added. For the active small-molecule donkey-hide gelatin mixture, the weight-average molecular weight ranges from 580 Da to 1300 Da, the peptide segments are distributed from 200 Da to 3000 Da, the dissolution rate in cold water is high, and the content of free amino acid is low; the active small-molecule donkey-hide gelatin mixture may be used for manufacturing small-molecule peptide food and health care food of donkey-hide gelatin.

Abstract: The subject of the present invention are peptide preparations obtained via the enzymatic digestion of hair, wool, bristles, animal fur and individual peptides with sequences corresponding to individual components of a peptide preparation with antitumour activity, for use in the treatment of tumours or oncological prophylaxis as basal components or components of compositions of substances for treating tumours or components of substances used in oncological prophylaxis.

Abstract: The present disclosure describes methods and systems for improving the expression of a properly folded, biologically active protein of interest in a cell free synthesis system. The methods and systems use a bacterial cell free extract having an active oxidative phosphorylation system, and include an exogenous protein chaperone. The exogenous protein chaperone can be expressed by the bacteria used to prepare the cell free extract. The exogenous protein chaperone can be a protein disulfide isomerase and/or a peptidyl-prolyl cis-trans isomerase. The inventors discovered that the combination of a protein disulfide isomerase and a peptidyl-prolyl cis-trans isomerase produces a synergistic increase in the amount of properly folded, biologically active protein of interest.

Abstract: The present invention provides a novel endo-?-N-acetylglucosaminidase (Endo-Om) using a transformant produced by cloning an endo-?-N-acetylglucosaminidase (Endo-Om) gene originated from a methylotrophic yeast Ogataea minuta IFO10746 strain. The Endo-Om according to the present invention has a specific activity 13-fold higher than that of known Endo-M and a Vmax value 55-fold higher than that of the known Endo-M, and is useful for the analysis of the structures of sugar chains, including complex type sugar chains, in glycoproteins and the modification of the sugar chains.

Abstract: To provide effective use from a yeast extract residue, an excess of which is produced as a byproduct of yeast extract, or reduction in amount of the yeast extract residue. Also to obtain various useful substances. A cell wall lytic enzyme having no protease is allowed to act on a yeast extract residue, after which a heat treatment is performed at 70 to 80° C. for 10 to 20 minutes, thereby enabling separation into a fraction of primarily cell walls and a fraction of primarily protein. A yeast protein having a protein content of 60% or more is obtained from the fraction of primarily protein and, by subjecting the yeast protein to enzymatic degradation, a seasoning having a high amount of total nitrogen is obtained.

Abstract: A compound may generally comprise the formula: wherein R1 is independently selected from C2-C10 alkyl or substituted alkyl, R2 is independently selected from the group consisting of —H and C1-C6 alkyl or substituted alkyl, X is selected from the group consisting of —NH— and —O—, Y is a carbohydrate, and m is an integer from 1 to 8. The compound may comprise a non-ionic acid labile surfactant. The compound may be used to facilitate solubilization of proteins and other molecules in an aqueous environment.

Abstract: The present invention includes compounds that are useful in perturbing or disrupting the function of a transmembrane or intracellular protein, whereby binding of a compound to the transmembrane or intracellular protein induces proteasomal degradation of the transmembrane or intracellular protein. The present invention further includes a method of inducing proteasomal degradation of a transmembrane or intracellular protein. The present invention further includes a method of identifying or validating a protein of interest as a therapeutic target for treatment of a disease state or condition.

Abstract: Disclosed herein is pyrroline-carboxy-lysine (PCL), a pyrrolysine analogue, which is a natural, biosynthetically generated amino acid, and methods for biosynthetically generating PCL. Also disclosed herein are proteins, polypeptides and peptides that have PCL incorporated therein and methods for incorporating PCL into such proteins, polypeptides and peptides. Also disclosed herein is the site-specific derivatization of proteins, polypeptides and peptides having PCL or pyrrolysine incorporated therein. Also disclosed herein is the crosslinking of proteins, polypeptides and peptides having PCL or pyrrolysine incorporated therein.

Abstract: Production of protein conjugate vaccines by use of transpeptidase enzymes, such as sortase enzymes. For example, homogenous immunoconjugates (e.g., a population of molecules having the same structure) formed by conjugating an antigenic polypeptide and a bacterial capsule component are provided. In certain aspects, methods for generating an immune response to B. anthracis by use of protective antigen-PDGA immunoconjugates are provided.

Abstract: The invention provides an endoglycosidase, referred to as EndoS49 and having the amino acid sequence of SEQ ID NO: 1. EndoS49 was isolated from Streptococcus pyogenes strain NZ131 and is a homolog of EndoS. EndoS49 has specific endoglycosidase activity on native IgG and cleaves a larger variety of Fc glycans than EndoS. A mutant thereof where the glutamic acid at position 186 of SEQ ID NO: 1 was substituted was produced: said mutant lacks endoalvcosidase activity but is capable of binding to IgG. Methods using EndoS49, deletions thereof and said mutant, especially for assessing glycosylation of IgG or for isolating IgG are disclosed.

Abstract: This invention describes a new analytical method to determine the quantity and distribution of fucose per Fc within an antibody preparation.

Abstract: Methods for producing and identifying fragments of proteins, and more particularly to methods for generating and identifying soluble protein domains are disclosed based on a method for generating a library of nucleic acid fragments from nucleic acid encoding a desired polypeptide, and more especially a library of essentially, randomly sampled fragments of coding DNA sequence predominantly of defined size range and a method for selecting cloned gene fragments from the library that encode soluble protein domains.

Abstract: The invention provides for genetically modified Bacillus host cells having decreased cytoplasmic reductase activity, such as decreased TrxA activity, and a nucleic acid encoding a heterologous Staphylococcal oxidase. The invention also provides for methods of producing di-sulfide bond containing proteins using the host cells of the invention and methods of improving protein folding using the host cell of the invention.

Abstract: The present invention relates to modified SAK gene having amino acid SEQ ID 2. The present invention further relates to process for cloning and expressing modified SAK gene fusion protein which imparts improved stability to the heterologous protein of interest. Further the invention relates to process of purification of recombinant heterologous proteins from bacterial inclusion bodies using modified SAK.

Abstract: Cell-free protein synthesis systems and methods of using the same for producing in vitro protein materials in high yield are disclosed. The cell-free protein synthesis platform includes (a) a Saccharomyces cerevisiae cellular extract prepared from mid-exponential to late-exponential batch cultures in the range from about 6 OD600 to about 18 OD600 or fed-batch cultures harvested in mid-exponential to late-exponential phase; (b) a reaction buffer; and (c) a translation template or (c?) a transcription template from which a translation template can be prepared in situ with an RNA polymerase. A method of performing high-throughput protein synthesis in vitro is also provided that utilizes a combined transcription/translation reaction with the cell-free protein synthesis platform from Saccharomyces cerevisiae, an RNA polymerase and a transcription template prepared from a source DNA using an amplification procedure.

Abstract: The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and/or the addition of a modifying group to a peptide.

Abstract: The invention relates to a process of preparing low molecular weight peptides from gelatine. The gelatine is broken down into low molecular weight peptides using enzymes from Bacillus sp. to obtain peptides with low ash content. The gelatine is clarified by adjusting the pH to 7 using alkali hydroxide and filtered. The filtered solution is desalted at 20 to 55° C. in a diafiltration mode till the salt content decreases below 0.05% (w/v) The product so obtained is hydrolysed with an enzyme maintaining pH in the range of 7 to 10 and at temperature in the range of 25 to 75° C. The hydrolysed product is filtered through ultrafiltration membrane at temperature in the range of 15 to 55° C. and permeate is concentrateds at temperature in the range of 50 to 100° C. under vacuum to obtain the peptide solution of the invention.

Abstract: The current disclosure provides binding polypeptides (e.g., antibodies), and effector moiety conjugates thereof (e.g., antibody-drug conjugates or ADCs), comprising a site-specifically engineered drug-glycan linkage within native or engineered glycans of the binding polypeptide. The current disclosure also provides nucleic acids encoding the antigen-binding polypeptides, recombinant expression vectors and host cells for making such antigen-binding polypeptides. Methods of using the antigen-binding polypeptides disclosed herein to treat disease are also provided.

Abstract: The present invention relates to kits and methods for efficiently generating 5? capped RNA having a modified cap nucleotide and for use of such modified-nucleotide-capped RNA molecules. In particular, the present invention provides kits and methods for capping RNA using a modified cap nucleotide and a capping enzyme system, such as poxvirus capping enzyme. The present invention finds use for in vitro production of 5?-capped RNA having a modified cap nucleotide and for in vitro or in vivo production of polypeptides by in vitro or in vivo translation of such modified-nucleotide-capped RNA. The invention also provides methods and kits for capturing or isolating uncapped RNA comprising primary RNA transcripts or RNA having a 5?-diphosphate, and methods and kits for using a capping enzyme system and modified cap nucleotides for labeling uncapped RNA comprising primary RNA transcripts or RNA having a 5?-diphosphate with detectable dye or enzyme moieties.

Abstract: The invention relates to a composition comprising a Vigna unguiculata seed extract. The composition is advantageously cosmetic, pharmaceutical, dermatological or nutraceutical. The invention further relates to a method for extracting a Vigna unguiculata seed extract, as well as to the resulting extract. The invention further relates to one such composition or one such extract for use in the prevention or treatment of disorders or pathologies of the skin, mucosae or appendages, for use in the prevention or treatment of vascular disorders, or for use in the prevention or treatment of adipose tissue alterations. Finally, the invention relates to a method for cosmetic care of the skin, appendages or mucosae, in order to improve the condition or appearance thereof, comprising the administration of one such composition or one such extract.

Abstract: Reagents and methods for the digital analysis of proteins or peptides are provided. Specifically provided herein are proteins for identifying the N-terminal amino acid or N-terminal phosphorylated amino acid of a polypeptide. Also, an enzyme for use in the cleavage step of the Edman degradation reaction and a method for using this enzyme are described.

Abstract: This invention relates to novel compounds, methods for selective chemical conjugation of protractor molecules and the use thereof for diagnostic and/or therapeutic purposes.

Abstract: Antibody variants having decreased or increased ability to mediate CDC due to modifications at the C-terminus of their heavy chains are described. Methods of generating such antibodies, as well as nucleotide constructs and host cells suitable for the production of said antibodies are also described.

Abstract: Methods of synthetically producing, formulating and using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 are provided. Methods of using secretoglobins SCGB1A1, SCGB3A2, and SCGB3A1 as therapeutic agents to affect long term patient outcomes, such as preventing severe respiratory exacerbations of underlying conditions that require medical intervention, including hospitalization are provided. Methods of producing recombinant human secretoglobins, analytical methods, pharmaceutical compositions, and methods of use to prevent the long term sequelae of acute and chronic respiratory conditions are provided.

Abstract: Disclosed is a Thermoanaerobacter sp. bacterial strain (BKHI) isolated from a hot spring, a purified protein (bioremediase) isolated from bacterial strain BKH1, as well as concrete compositions comprising BKH1 and/or the protein, and methods of using the protein and/or composition. Also disclosed are nucleic acids encoding the protein isolated from BKHI, as well as expression vectors, host cells, cell lines, and methods for generating and purifying the bioremediase protein.

Abstract: The invention provides a method of incorporating nonstandard amino acids into a protein by utilizing a modified aminoacyl-tRNA synthetase to charge the nonstandard amino acid to a modified tRNA, which forms strict Watson-Crick base-pairing with a codon that normally forms wobble base-pairing with natural tRNAs.

Abstract: The present invention related to a method of producing polypeptides in transformed host cells by expressing a precursor molecule of the desired polypeptide which are to be acylated in a subsequent in vitro step. The invention is also related to DNA-sequences, vectors and transformed host cells for use in the claimed method. Further, the present invention is related to certain precursors of the desired polypeptides and certain acylation methods. The invention provides a method for making polypeptides being preferentially acylated in certain lysine ?-amino groups.

Abstract: The invention relates to a novel 3D structure encoding a Nocardiopsis protease, as well as to variants of parent protease homologous to Nocardiopsis proteases, preferably of improved thermostability and/or with an amended temperature activity profile. The invention also relates to DNA sequences encoding such variants, their production in a recombinant host cell, as well as methods of using the variants, in particular within the field of animal feed and detergents. The invention furthermore relates to methods of generating and preparing protease variants of amended properties.

Abstract: An in vitro process of preparing virus-like particles (VLPs) from recombinant papaya mosaic virus coat protein and ssRNA, which allows for large scale production of VLPs in high yields, is provided. Also provided are VLPs comprising ssRNA prepared by the in vitro process. The VLPs can be used as adjuvants and when fused to an antigen, as vaccines. The use of the VLPs for stimulation of the innate immune response is also provided.

Abstract: The present invention is directed to a cell-free system for producing a glycosylated protein. This system comprises an isolated oligosaccharyltransferase capable of transferring a glycan from a lipid carrier molecule to a glycoprotein target, one or more isolated glycans, where each glycan is linked to a lipid carrier molecule, and a glycoprotein target comprising one or more glycan acceptor amino acid residues or a nucleic acid molecule encoding said glycoprotein target. The present invention further relates to kits and methods for producing a glycosylated protein in this cell-free system.

Abstract: Disclosed herein are methods of treating a subject with pulmonary disease including administering to the subject a therapeutically effective amount of a polypeptide including at least one arginine residue susceptible to ADP-ribosylation and nicotinamide adenine dinucleotide (NAD). In some embodiments, the polypeptide and/or NAD is administered via inhalation. Also disclosed is a pharmaceutical composition including at least one polypeptide (such as HNP-1) and NAD. The disclosure also provides in vitro methods of producing a polypeptide with altered activity, including contacting the polypeptide with NAD and an arginine-specific mono-ADP-ribosyltransferase (for example, ART1) to produce a polypeptide including at least one ADP-ribosylated arginine residue, incubating the ADP-ribosylated polypeptide under conditions sufficient for conversion of at least one ADP-ribosylated arginine residue to ornithine, and isolating the ornithine-containing polypeptide.

Abstract: Methods and reagents for the installation of click chemistry handles on target proteins are provided, as well as modified proteins comprising click chemistry handles. Further, chimeric proteins, for example, bi-specific antibodies, that comprise two proteins conjugated via click chemistry, as well as methods for their generation and use are disclosed herein.

Abstract: The present invention provides methods of refolding mammalian glycosyltransferases that have been produced in bacterial cells, and methods to use such refolded glycosyltransferases, including glycosyltransferase mutants that have enhanced ability to be refolded. The invention also provides methods of refolding more than one glycosyltransferase in a single vessel, methods to use such refolded glycosyltransferases, and reaction mixtures comprising the refolded glycosyltransferases.

Abstract: This invention pertains to methods for producing homogeneous recombinant proteins that contain polymer initiators at defined sites. The unnatural amino acid, 4-(2?-bromoisobutyramido)phenylalanine of formula 1, was designed and synthesized as a molecule comprising a functional group further comprising an initiator for an atom-transfer radical polymerization (‘ATRP”) that additionally would provide a stable linkage between the protein and growing polymer. We evolved a Methanococcus jannaschii (Mj) tyrosyl-tRNA synthetase/tRNACUA pair to genetically encode this unnatural amino acid in response to an amber codon. To demonstrate the utility of this functional amino acid, we produced Green Fluorescent Protein with the unnatural amino acid initiator of formula 1 site-specifically incorporated on its surface (GFP-1).

Abstract: This invention provides compositions and methods for producing translational components that expand the number of genetically encoded amino acids in eukaryotic cells. The components include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, orthogonal pairs of tRNAs/synthetases and unnatural amino acids. Proteins and methods of producing proteins with unnatural amino acids in eukaryotic cells are also provided.

Abstract: Provided herein are, inter alia, methods for linking an mRNA molecule to a polypeptide (e.g., a peptide or a protein) by linking the mRNA molecule to a linking amino acid in the polypeptide, or by linking the mRNA molecule to a linking tRNA to which the polypeptide is attached, via reactions not catalyzed by the ribosome, and methods for making polypeptide libraries. Also provided are mRNA-protein complexes and mRNA-tRNA-protein complexes, libraries containing these complexes, and methods of using these complexes.

Abstract: The current invention comprises a method for producing an immunoglobulin or immunoglobulin fragment or immunoglobulin fusion with G1 glycostructure comprising incubating with a galactosyltransferase, a sialyltransferase, a beta-1,4-galactosidase and a sialidase, whereby the galactosyltransferase is added in more than one aliquot during the incubating.

Abstract: The present invention relates to isolated polypeptides having protease activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: There is provided a method for producing free polyubiquitin chains linked through a single desired lysine residue, comprising the steps of: (a) selecting an E3 ubiquitin ligase enzyme which is homologous to mammalian HECT E3 ligases and possesses the desired lysine residue specificity; (b) incubating the E3 enzyme with an E1 ubiquitin activating enzyme, an E2 ubiquitin conjugating enzyme and monomeric ubiquitin; and (c) if undesired linkages are present, removing the undesired linkages by exposure to a DUB enzyme having the appropriate specificity.

Abstract: A method for the production of hydrolyzed allergens from allergens comprising the steps of: a) extracting a source of allergens comprising allergenic proteins to form an extract, b) purifying the extract to remove non-protein components to form a purified extract, c) denaturing the purified extract with a first denaturing agent to form a purified denatured extract, d) refining the purified denatured extract to remove impurities to form a refined denatured extract, e) denaturing the refined denatured extract with a second denaturing agent to form denatured allergen mixture, and f) hydrolyzing the denatured allergen mixture to form the hydrolyzed allergens.

Abstract: The present invention relates to kits and methods for efficiently generating 5? capped RNA having a modified cap nucleotide and for use of such modified-nucleotide-capped RNA molecules. In particular, the present invention provides kits and methods for capping RNA using a modified cap nucleotide and a capping enzyme system, such as poxvirus capping enzyme. The present invention finds use for in vitro production of 5?-capped RNA having a modified cap nucleotide and for in vitro or in vivo production of polypeptides by in vitro or in vivo translation of such modified-nucleotide-capped RNA. The invention also provides methods and kits for capturing or isolating uncapped RNA comprising primary RNA transcripts or RNA having a 5?-diphosphate, and methods and kits for using a capping enzyme system and modified cap nucleotides for labeling uncapped RNA comprising primary RNA transcripts or RNA having a 5?-diphosphate with detectable dye or enzyme moieties.