Genetically engineered bacteria, pharmaceutical compositions thereof, and methods of modulating and treating diseases associated with hyperphenylalaninemia are disclosed.

Provided herein are recombinant polynucleotides including a first nucleic acid sequence encoding an antigen, and a second nucleic acid sequence encoding an enzyme of an amino acid catabolic pathway. The provided recombinant polynucleotides are particularly useful for inducing long-lived immune responses having improved memory characteristics. Also provided are pharmaceutical compositions, viral particles, and host cells including the disclosed recombinant polynucleotides, and methods for using the disclosed materials.

The invention is directed to methods of promoting myelin formation in central nervous system (CNS) tissue in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of interleukin-4 induced gene 1 (IL4-i1) protein. The invention is also directed to methods of monitoring the progression of conditions marked by an impairment of myelin formation in the CNS comprising assessing the levels or activity of IL4-i1 in activated macrophages obtained from the subject.

Aspects of the present disclosure are drawn to methods of improving the expression of secreted cuproenzymes from host cells by manipulating the expression level of one or more proteins involved in copper transport in the host cell, e.g., membrane-bound copper transporting ATPases and soluble copper transporters. The present disclosure also provides compositions containing such improved host cells as well as products derived from the improved host cells that contain one or more cuproenzymes of interest.

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells comprising a heterologous gene encoding a branched chain amino acid catabolism enzyme. The disclosure further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders involving the catabolism of branched chain amino acids using the pharmaceutical compositions disclosed herein.

The present disclosure discloses a genetically engineered strain, belonging to the technical field of bioengineering. L-amino acid oxidase genes, α-keto acid decarboxylase genes, alcohol dehydrogenase genes, and enzyme genes capable of reducing NAD(P) to NAD(P)H are introduced into the genetically engineered strain of the present disclosure. The present disclosure further discloses a construction method and application of a recombinant Escherichia coli genetically engineered strain. When being applied to the biosynthesis of phenylethanoids, the method of the present disclosure has the characteristics of simple operation, low cost, and high synthesis efficiency and optical purity of the product, and has good industrialization prospects.

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells comprising a heterologous gene encoding a branched chain amino acid catabolism enzyme. The disclosure further provides pharmaceutical compositions comprising the recombinant bacteria, and methods for treating disorders involving the catabolism of branched chain amino acids using the pharmaceutical compositions disclosed herein.

The disclosure discloses preparation of an L-amino acid deaminase mutant and application thereof, belonging to the technical field of gene engineering. Through pmirLAAD protein modification, analysis of a flexible loop structure around a binding site of the pmirLAAD product, and design of the best mutant, the modification method of the disclosure overcomes the defect that the catalytic efficiency of the previous wild-type enzyme is reduced due to product inhibition, and is tested by experiments. Compared with the control, the catalytic efficiency (1.61 mM.sup.−1.Math.min.sup.−1) and the product inhibition constant (5.4 mM) of the finally obtained best mutant pmirLAAD.sup.M4 are respectively increased by 5.2 times and 5.7 times. The yield of α-ketoisovaleric acid can reach 96.5 g/L, and the transformation rate is greater than 97%. By adopting the method of the disclosure, the cost can be greatly reduced, and the industrialization process of production of α-ketoisovaleric acid by an enzymatic transformation method is accelerated.

Genetically engineered microorganisms, e.g., genetically engineered bacteria, compositions and formulations thereof, as well as methods for characterizing, dosing, and determining the activity of the bacteria, compositions, and formulations, e.g., using a live cell counting method are disclosed.

Provided is a use of an agent for regulating an amino acid level in preparing a drug for treating Alzheimer's disease in a subject.