The invention provides an effective method for improving N-acetylglucosamine (GlcNAc) production by engineered B. subtilis Deletion of phosphoenolpyruvate carboxykinase encoding gene pckA and encoding pyruvate kinase gene pyK in recombinant GlcNAc-producing strain BSGNK-PxylA-glmS-P43-GNA1 (BSGNK) is first performed to enhance GlcNAc production, followed by overexpression of pyruvate carboxylase encoding gene pycA for facilitating cell growth. Finally, the GlcNAc production of the recombinant strain BPTS3 reached to 11.3 g/L, which was 1.84-fold of BSGNK. This method can be used for improve cellular property of engineered B. subtilis for GlcNAc production, which can be further applied to industrial production of GlcNAc.

The present disclosure provides polynucleotide cassettes, expression vectors and methods for the expression of a gene in mammalian cells to provide gene therapy for pyruvate kinase deficiency.

The present disclosure relates, in some aspects, to cell-free methods and systems for large-scale conversion of methane to isobutanol, comprising combining, in a bioreactor at elevated pressure, methane, oxygen, and cell lysates containing methane monooxygenase, methanol dehydrogenase, and enzymes that catalyze the conversion of formaldehyde to isobutanol, to form a cell-free reaction mixture, and incubating under suitable conditions the cell-free reaction to convert methane to isobutanol.

The invention provides an effective method for improving N-acetylglucosamine (GlcNAc) production by engineered B. subtilis Deletion of phosphoenolpyruvate carboxykinase encoding gene pckA and encoding pyruvate kinase gene pyK in recombinant GlcNAc-producing strain BSGNK-PxylA-glmS-P43-GNA1 (BSGNK) is first performed to enhance GlcNAc production, followed by overexpression of pyruvate carboxylase encoding gene pycA for facilitating cell growth. Finally, the GlcNAc production of the recombinant strain BPTS3 reached to 11.3 g/L, which was 1.84-fold of BSGNK. This method can be used for improve cellular property of engineered B. subtilis for GlcNAc production, which can be further applied to industrial production of GlcNAc.

The present disclosure relates, in some aspects, to cell-free methods and systems for large-scale conversion of methane to isobutanol, comprising combining, in a bioreactor at elevated pressure, methane, oxygen, and cell lysates containing methane monooxygenase, methanol dehydrogenase, and enzymes that catalyze the conversion of formaldehyde to isobutanol, to form a cell-free reaction mixture, and incubating under suitable conditions the cell-free reaction to convert methane to isobutanol.

Described herein are methods of treating pyruvate kinase deficiency (PKD), sickle cell disease or thalassemia with mitapivat or a pharmaceutically acceptable salt thereof, or use of the drug for the treatment of these conditions, in combination with or in the absence of with a secondary drug, such as an inducer or an inhibitor of cytochrome P450. Various doses and dosing regimens of mitapivat in monotherapy and in concomitant medications are described.

The present disclosure relates to genetically engineered hematopoietic cell/s, specifically, lymphocytes, and more specifically, cells of the T cell lineage or a cell population comprising at least one of the cell/s. The disclosed cells comprises and/or expresses at least one nucleic acid sequence encoding at least one molecule involved directly or indirectly in at least one metabolic pathway. The present disclosure provides compositions, methods and uses of the engineered cells.

Quantum dots (QDs) and nanoplatelets (NPLs) are two types of nanoparticles used as scaffolds for enzymes operating in enzymatic cascades. Combinations of QDs and NPLs were surprisingly found to operate synergistically to create a greater enhancement than either alone when operating as scaffolds for enzymatic cascade reactions. A process involves providing an enzymatic cascade including a cluster of nanoparticles including both QDs and NPLs and having a plurality of enzymes bound thereto, the enzymes configured as an enzymatic cascade, such that the product of a first enzyme is a substrate of a second enzyme; contacting the cascade cluster with a substrate of the first enzyme; and allowing a reaction to proceed so that each of the plurality of enzymes acts in succession to produce an end product. The enzymes are bound to the nanoparticles via metal affinity coordination between histidine tags on the enzymes and zinc-containing surfaces of the nanoparticles.

The present disclosure provides polynucleotide cassettes, expression vectors and methods for the expression of a gene in mammalian cells to provide gene therapy for pyruvate kinase deficiency.

Provided herein are methods for diagnosing and treating Alzheimer's disease in a subject comprising determining the expression level of three, four or five members of a panel of proteins in a biological sample obtained from the subject.