Disclosed herein is a method for expressing phytase in a filamentous fungus by using an optimized Escherichia coli phytase gene having a nucleotide sequence as shown in SEQ ID NO. 7 and a signal peptide having a nucleotide sequence as shown in SEQ ID NO. 12.

The present invention generally relates to methods of activating cells for use in therapy. For example, the invention relates to preparing cells ex vivo for use in immunotherapy, particularly cancer immunotherapy. More specifically, the invention relates to methods for the preparation of leukocytes, particularly T cells through PTP1B inhibition, exhibiting cytotoxic properties for use in adoptive cell transfer. The invention also relates to cells and compositions including them for cancer immunotherapy. The invention also relates to methods of immunotherapy, particularly cancer immunotherapy.

A process for producing a protein isolate from an oilseed meal, and the isolate thus obtained, said isolate comprising proteins and an amount of 4 wt. % or less of phytic acid, said amount of phytic acid being by weight of proteins in said isolate. The process may comprise the following steps: a) providing an oilseed meal; b) mixing the oilseed meal with a first aqueous solvent to form a slurry at a pH ranging from 6 to 7.8, said slurry having a solid phase; c) separating said solid phase from said slurry, d) mixing said separated solid phase with a second aqueous solvent at a pH ranging from 1 to 3.5, preferably from 2 to 3, to form a mixture said mixture having a liquid phase; e) separating said liquid phase from said mixture formed in step d); f) f1) mixing the separated liquid phase to a phytase at a temperature and a pH suitable for phytase activity to obtain a mixture having a liquid phase and a solid phase; and/or f2) mixing the separated liquid to a salt, to obtain a resulting liquid composition having a molar concentration of said salt ranging from 0.05M to 0.5M, at a temperature ranging from 40 C. to 70 C., to obtain a mixture having a liquid phase and a solid phase; g) precipitating a solid phase from the liquid of step f) for example by a cooling down step of the mixture to a temperature of 30 C. or less; h) separating said solid precipitate from the liquid of step g) said liquid comprising a water-rich liquid phase and an oil-rich liquid phase; i) separating said water-rich liquid phase from said oil-rich liquid phase, j) subjecting said water-rich liquid phase obtained in step i) to one or several membrane filtration(s) to obtain a protein isolate; and k) optionally, drying said protein isolate to obtain a dry protein isolate.

Novel cell penetrating agents for intracellular delivery of desired cargo, including proteins. Use of cell penetrating agents to deliver cargos to the interior of cells and cellular compartments and organelles is transformative for diagnostic, therapeutic, and research processes.

The present disclosure relates to methods for using one or more polypeptides with phytase activity in grain processing, ethanol, and biofuel production.

Methods and compositions are described for producing a phytase in transgenic maize plants and then incorporating parts of the transgenic maize plants in animal feed. The feed phytase enzyme displays activity across a broad pH range, and tolerance to temperatures that are often encountered during the process of preparing animal feeds. Methods of producing an animal feed that incorporate the transgenic maize plants, parts thereof or plant derived phytases, as well as methods of promoting the release of inorganic phosphate from a phytic acid in an animal, producing an animal meat, or reducing the ratio of intake of an animal feed per weight of the animal meat by feeding an animal with the animal feed incorporating transgenic maize plants are provided.

The present invention provides enzymes that have been optimized by implementation of Protein Repair One Stop Shop (PROSS), an algorithm that generates protein design(s) for enhanced stability without changing either enzymatic properties or enzyme active site conformation of the respective enzyme. The protein design(s) generated by PROSS introduce mutations to the amino acid sequence of a wild-type protein, resulting in a mutated amino acid sequence that encodes a variant of the wild-type enzyme, i.e., an enzyme variant, which has an enhanced stability, core packing, surface polarity and backbone rigidity, a higher functional expression, and/or a combination thereof, compared to the stability core packing, surface polarity and backbone rigidity, functional expression and/or a combination thereof, of the wild-type enzyme.

Methods for enhancing phytase thermal stability by fusing binding elements to target phytases are provided. Engineered phytases that include binding elements fused to target phytases to cause cyclization of the engineered phytases and enhance thermal stability of the target phytases are described. Engineered nucleic acids encoding engineered phytases and hosts engineered to express engineered nucleic acids are also provided. Methods for incorporating engineered phytases in animal feed and animal feed including the same are described.

The present invention relates to the field of genetic engineering, particularly to phytase variants YkAPPA having amino acid sequence substituting Leucine at the 162.sup.th site of the sequence set forth in SEQ ID NO.1 with glycine or proline, or having amino acid sequence substituting glutamic acid at the 230.sup.th site of the sequence set forth in SEQ ID NO.1 with glycine, alanine, serine, threonine, aspartic acid, proline, or arginine, and having improved pepsin resistance and increased catalytic efficiency of 2.1 times of that of the wild phytase, in the benefit of the development of economical feed enzyme industry.

The present disclosure provides recombinant microorganisms and methods for the production of 4-HMF, 2,4-furandimethanol, furan-2,4-dicarbaldehyde, 4-(hydroxymethyl)furoic acid, 2-formylfuran-4-carboxylate, 4-formylfuran-2-carboxylate, and/or 2,4-FDCA from a carbon source. The method provides for engineered microorganisms that express endogenous and/or exogenous nucleic acid molecules that catalyze the conversion of a carbon source into 4-HMF, 2,4-furandimethanol, furan-2,4-dicarbaldehyde, 4-(hydroxymethyl)furoic acid, 2-formylfuran-4-carboxylate, 4-formylfuran-2-carboxylate, and/or 2,4-FDCA. The disclosure further provides methods of producing polymers derived from 4-HMF, 2,4-furandimethanol, furan-2,4-dicarbaldehyde, 4-(hydroxymethyl)furoic acid, 2-formylfuran-4-carboxylate, 4-formylfuran-2-carboxylate, and/or 2,4-FDCA.