This invention relates to genetically engineered strains of yeast and methods for producing recombinant protein (e.g., collagen). Recombinant protein of the present invention is used to produce biofabricated leather or a material having leather-like properties containing recombinant or engineered collagen. The yeast strains are engineered to produce ascorbate and/or increased production of ketoglutarate.

This invention relates to genetically engineered strains of yeast and methods, for producing recombinant protein (e.g., collagen). Recombinant protein of the present invention is used to produce biofabricated leather or a material having leather-like properties containing recombinant or engineered collagen. The yeast strains are engineered to produce ascorbate and/or increased production of ketoglutarate.

The inventors previously demonstrated that mitochondrial VDAC1 directly induces Schwann cell demyelination via MAPK and c-jun activation after sciatic nerve injury and diabetic neuropathy and CMT1A. They found that reduction of mitochondrial calcium release by VDAC1 blocking strongly reduces the number of demyelinating Schwann cell in vivo and improve nerve conduction and neuromuscular activity in diabetic, Guillain-Barre syndrome and Charcot-Marie Tooth disease models. Herein, the inventors precisely map the binding region of the N-terminal HK-1 helix through an ala scan completed by a deletion study. Furthermore, they optimized the HK-derived peptide through stabilization of the helix by replacement of non-essential amino acids by the a-aminoisobutyric acid (Aib) known as a helix inducer. Additionally, they described an in-house cellular screening assay based on the ability of MJ to detach HK from VDAC that allows to determine the peptide potency. Overall, their data confirm that N-terminal HK derived peptides acting on VDAC are promising tools for the study of the demyelination process. Thus, the present invention refers to optimized HK-derived peptide and its use for treating peripheral demyelinating disease, myocardium diseases.sup.10 11, cancer.sup.12,13-15, diabetes.sup.14 14-16, lupus-like diseases.sup.17, non-alcoholic fatty liver disease.sup.24,25, chemoinduced neuropathy9 Alzheimer disease.sup.18 19, Parkinson disease.sup.20, Huntington disease.sup.21, ALS.sup.22,23 and more generally all neurodegenerative diseases linked to a protein aggregation.sup.28.

Object of the invention is a synthetic peptide, in particular a synthetic peptide to be used as medicament, in particular to be used in the treatment of neurodegenerative diseases and Amyotrophic Lateral Sclerosis (ALS), and compositions comprising such synthetic peptide. Furthermore, the invention concerns processes for the preparation of said synthetic peptide.

A reagent for glutamine synthetase reaction comprising a chelating agent and glutamine synthetase, and a reagent for quantification of ammonia comprising a chelating agent, ATP, glutamic acid, glutamine synthetase, glucose, an oxidized NAD compound, ADP-dependent hexokinase, and glucose-6-phosphate dehydrogenase, are provided.

The present invention discloses anti-cancer compositions, and associated methods, including an anti-cancer composition comprising: a cellular energy inhibitor having the structure according to formula I

##STR00001## wherein X is selected from the group consisting of: a nitro, an imidazole, a halide, sulfonate, a carboxylate, an alkoxide, and amine oxide; and R is selected from the group consisting of: OR, N(R).sub.2, C(O)R, C1-C6 alkyl, C6-C12 aryl, C1-C6 heteroalkyl, a C6-C12 heteroaryl, H, and an alkali metal; where R represents H, alkali metal, C1-C6 alkyl, C6-C12 aryl or C(O)R, R represents H, C1-C6 alkyl, or C6-C12 aryl, and R represents H, C1-C20 alkyl or C6-C12 aryl. The anti-cancer composition can additionally comprise at least one sugar, which stabilizes the cellular energy inhibitor by substantially preventing the inhibitor from hydrolyzing. Also, the anti-cancer composition can comprise a hexokinase inhibitor. Further, the anti-cancer composition can comprise a biological buffer that is present in an amount sufficient to at least partially deacidify the cellular energy inhibitor and neutralize metabolic by-products of the cellular energy inhibitor.

Compositions and methods for protecting a subject against, or treating a subject with, a pathogenic infection are presented. A method includes administering a composition including a cellular energy inhibitor having the structure according to formula I

##STR00001## wherein X is selected from the group consisting of: a nitro, an imidazole, a halide, sulfonate, a carboxylate, an alkoxide, and amine oxide; and R is selected from the group consisting of: OR, N(R).sub.2, C(O)R, C1-C6 alkyl, C6-C12 aryl, C1-C6 heteroalkyl, a C6-C12 heteroaryl, H, and an alkali metal; where R represents H, alkali metal, C1-C6 alkyl, C6-C12 aryl or C(O)R, R represents H, C1-C6 alkyl, or C6-C12 aryl, and R represents H, C1-C20 alkyl or C6-C12 aryl, at least one sugar to stabilize the cellular energy inhibitor and a biological buffer present in an amount sufficient to at least partially deacidify the cellular energy inhibitor and neutralize metabolic by-products of the cellular energy inhibitor.

The present invention relates to a process of preparing a rubber-like material containing a protein immobilized therein, as well as a corresponding rubber-like material, the process comprising the steps of (a) mixing a protein, a branched polymer such as trimethylolpropane ethoxylate and a linear polymer such as poly(ethylene oxide) in an aqueous solution to form a gel, and (b) drying the gel to obtain a rubber-like material containing the protein immobilized therein, wherein the branched polymer comprises at least three polymeric branches bound to a central branching unit. The rubber-like material allows the immobilization and stabilization of a wide range of different proteins, including luminescent proteins as well as enzymes, and can particularly advantageously be used as down-converting material for light-emitting diodes (LEDs), for diagnostic applications, and in bioreactors.

Disclosed herein are genetically engineered yeast cells capable of producing lactate from sucrose. The genetically engineered yeast cells comprise a polynucleotide encoding an exogenous lactate dehydrogenase enzyme: a polynucleotide encoding an exogenous invertase enzyme: a deletion or disruption of a native pyruvate decarboxylase (PDC) gene: and a genetic modification resulting in overexpression of a native hexokinase gene.

This disclosure provides a composition containing a conjugate with a modified insulin molecule. The conjugate has an insulin molecule, which can be insulin or an insulin analog, glucagon, GLP-1, GLP-2 or a GLP-1 agonist. The conjugate also contains one or more polymers. Each of the one or more polymers is covalently linked to the insulin molecule. Additionally, each of the one or more polymers is covalently linked to between 0 to 50 copies of a decoy ligand, and to between 0 to 50 copies of a glucose-binding agent, such that the combined total number of glucose-binding agents and decoy ligands covalently linked to each of the one or more polymers is at least 1. The conjugate can reversibly bind to soluble glucose and in which the extent of its glucose-binding controls the extent to which the modified insulin is able to bind to and activate the insulin receptor. Methods of making the conjugate, as well as use of the conjugate in treatment, are also provided.