Patent classifications
C12N9/06
Thermostabilized Amadoriases and uses thereof
The present invention refers to Amadoriase enzyme protein variants having de-glycating activity and improved thermostability compared to the wild type Amadoriase. The present invention refers also to the use of the thermostabilized Amadoriase as deglycating agent, preferably in the food industry. Moreover, the present invention refers to the use of the thermostabilized Amadoriase as diagnostic and/or therapeutic tools. Preferably, the Amadoriase enzyme protein variants of the invention can be used for determining the level of glycated haemoglobin in a biological sample and therefore for monitoring diabetes.
In vitro glycosylation of proteins and enzymes
The present invention is broadly concerned with new in vitro glycosylation methods that provide rational approaches for producing glycosylated proteins, and the use of glycosylated proteins. In more detail, the present invention comprises methods of glycosylating a starting protein having an amino sidechain with a nucleophilic moiety, comprising the step of reacting the protein with a carbohydrate having an oxazoline moiety on the reducing end thereof, to covalently bond the amino sidechain of the starting protein with the oxazoline moiety, wherein the glycosylated protein substantially retains the structure and function of the starting protein. Target proteins include oxidase, oxidoreductase and dehydrogenase enzymes. The glycosylated proteins advantageously have molecular weights of at least about 7500 Daltons. In a further embodiment, the present invention concerns the use of glycosylated proteins, fabricated by the methods disclosed herein, in the assembly of amperometric biosensors.
MICROORGANISMS AND METHODS FOR ENHANCING THE AVAILABILITY OF REDUCING EQUIVALENTS IN THE PRESENCE OF METHANOL, AND FOR PRODUCING 3-HYDROXYISOBUTYRATE OR METHACRYLIC ACID RELATED THERETO
Provided herein is a non-naturally occurring microbial organism having a methanol metabolic pathway that can enhance the availability of reducing equivalents in the presence of methanol. Such reducing equivalents can be used to increase the product yield of organic compounds produced by the microbial organism, such as 3-hydroxyisobutyrate or MAA. Also provided herein are methods for using such an organism to produce 3-hydroxyisobutyrate or MAA.
METHODS OF TREATING VIRAL INFECTIONS AND HEALTH CONSEQUENCES
This invention relates to formulations of uric acid lowering agent(s) (UALA) designed to inhibit xanthine oxidase and/or decrease serum or tissue uric acid concentration for the treatment and prevention of morbidities and mortality during viral infection. For example, acute kidney injury due to coronavirus infection by administering a therapeutically effective amount of an agent capable of inhibiting xanthine oxidase and/or decreasing uric acid levels in a patient in need of such treatment. Additionally, the scope of the invention includes a method of treating and preventing acute kidney injury and health consequences due to coronavirus infection.
L-GLUTAMATE OXIDASE MUTANT
The present invention provides an alternative L-glutamate oxidase that allows for measurement of L-glutamate. More specifically, the present invention provides the following L-glutamate oxidase mutant (a) or (b) and the like: (a) an L-glutamate oxidase mutant including an amino acid sequence that has 90% or more identity to an amino acid sequence of SEQ ID NO: 3 and exhibits an activity of oxidizing L-glutamate, except an L-glutamate oxidase including an amino acid sequence of SEQ ID NO: 1; or (b) an L-glutamate oxidase mutant comprising a peptide linker consisting of 1 to 20 amino acid residues which is inserted into one or more sites selected from the group consisting of (1) a site in a region proximity to a boundary between α1 and α2 regions, (2) a site in a region proximity to a boundary between α2 and γ regions and (3) a site in a region proximity to a boundary between γ and β regions in the L-glutamate oxidase mutant (a), and having the activity of oxidizing L-glutamate.
TRANSGLUTAMINASE VARIANTS AND APPLICATIONS OF USE THEREOF
Variants of the transglutaminase enzyme of Streptomyces mobaraensis are provided. The disclosed variants exhibit at least about 2-fold increased enzymatic activity versus the wild-type enzyme. Methods and compositions are provided for biocidal applications of use and for covalently binding small organic molecules to a protein or material of interest. Methods are provided for increasing the self-life of products, such as personal care, household and industrial products, by incorporating an effective amount of the disclosed variant enzymes into the product. The transglutaminase variants may also be used to covalently bind functional ingredients, such as UV-blocking molecules, dyes, or pigments to proteins. The transglutaminase enzymes and functional ingredients may be incorporated into a cosmetic formulation for modifying skin, hair, or nail proteins or skin-derived proteins, such as collagen, keratin, and/or elastin.
AMADORIASE HAVING ENHANCED DEHYDROGENASE ACTIVITY
Provided are an amadoriase that is less likely to be influenced by oxygen concentration and a method and a reagent kit for measurement of HbA1c using such amadoriase. Provided are an amadoriase that is obtained via substitution of one or more amino acid residues at a position or positions corresponding to the position(s) selected from the group consisting of positions 280, 267, 269, 54, and 241 of the amadoriase derived from the genus Coniochaeta, a method for measurement of HbA1c, a reagent kit for measurement, and a sensor using such amadoriase. The modified amadoriase according to the invention has a lowered oxidase activity and an enhanced dehydrogenase activity, and this enables the use of an electron mediator, and this reduces the influence of oxygen concentration. Thus, HbA1c can be measured with high sensitivity.
ENGINEERED IMINE REDUCTASES AND METHODS FOR THE REDUCTIVE AMINATION OF KETONE AND AMINE COMPOUNDS
The present application provides engineered polypeptides having imine reductase activity, polynucleotides encoding the engineered imine reductases, host cells capable of expressing the engineered imine reductases, and methods of using these engineered polypeptides with a range of ketone and amine substrate compounds to prepare secondary and tertiary amine product compounds.
Method for producing objective substance
A method for producing an objective substance such as vanillin and vanillic acid is provided. An objective substance is produced from a carbon source or a precursor of the objective substance by using a microorganism that is able to produce the objective substance, which microorganism has been modified so that the activity of an enzyme involved in SAM cycle (SAM cycle enzyme) is increased.
Purification and isolation of recombinant oxalate degrading enzymes and spray-dried particles containing oxalate degrading enzymes
The present invention comprises methods and compositions for the reduction of oxalate in humans, and methods for the purification and isolation of recombinant oxalate reducing enzyme proteins. The invention provides methods and compositions for the delivery of oxalate-reducing enzymes in particle compositions. The compositions of the present invention are suitable in methods of treatment or prevention of oxalate-related conditions.