This disclosure relates to the engineering of phototrophic microorganisms for conversion of alkanes into higher-value products. Recombinant phototrophic organisms such as cyanobacteria can be engineered, optionally in a modular format, to express enzymes involved in converting methane to methanol, methanol to formaldehyde, formaldehyde to central metabolic pathway intermediates, and such intermediates to n-butanol.

Described herein are fusion proteins including methanol dehydrogenase (MeDH) and at least one other polypeptide such as 3-hexulose-6-phosphate dehydrogenase (HPS) or 6-phospho-3-hexuloisomerase (PHI), such as DHAS synthase or fructose-6-Phosphate aldolase or such as DHA synthase or DHA kinase. In a localized manner, the fusion protein can promote the conversion of methanol to formaldehyde and then to a ketose phosphate such as hexulose 6-phosphate or then to DHA and G3P. When expressed in cells, the fusion proteins can promote methanol uptake and rapid conversion to the ketose phosphate or to the DHA and D3P, which in turn can be used in a pathway for the production of a desired bioproduct. Beneficially, the rapid conversion to the ketose phosphate or to the DHA and G3P can avoid the undesirable accumulation of formaldehyde in the cell. Also described are engineered cells expressing the fusion protein, optionally include one or more additional metabolic pathway transgene(s), methanol metabolic pathway genes, target product pathway genes, cell culture compositions including the cells, methods for promoting production of the target product or intermediate thereof from the cells, compositions including the target product or intermediate, and products made from the target product or intermediate.

An object of the present invention is to provide a series of techniques for producing 1,4-butanediol from methanol or the like. Provided is a recombinant cell prepared by introducing a gene encoding at least one enzyme selected from the group consisting of succinate semialdehyde dehydrogenase, succinyl-CoA synthase, CoA-dependent succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, 4-hydroxybutyryl-CoA transferase, 4-hydroxybutyryl-CoA reductase, 4-hydroxybutyraldehyde dehydrogenase, and alcohol dehydrogenase, into a host cell which is a methylotroph, wherein the gene is expressed in the host cell, and the recombinant cell is capable of producing 1,4-butanediol from at least one C1 compound selected from the group consisting of methane, methanol, methylamine, formic acid, formaldehyde, and formamide.

The present disclosure provides compositions, methods of use, and methods of creation for a population of transgenic plants derived from plant cells transformed with recombinant DNA for expression of heterologous proteins. In particular, the present disclosure provides compositions comprising indoor ornamental plants suited for the removal of volatile organic compounds such as formaldehyde, benzene, toluene, ethylbenzene and/or xylene from air. Also disclosed are transgenic seeds for growing a transgenic plant having the recombinant DNA in its genome and exhibiting enhanced VOC removal from air. Also disclosed are methods for generating seed and plants based on the transgenic events. Also disclosed are microbes selected for during directed evolution to have enhanced VOC removal from air capabilities. Also disclosed are methods and compositions for generating plant-microbiome pairings for enhanced VOC removal from air.

A combination of an electrochemical device for delivering reducing equivalents to a cell, and engineered metabolic pathways within the cell capable of utilizing the electrochemically provided reducing equivalents is disclosed. Such a combination allows the production of commodity chemicals by fermentation to proceed with increased carbon efficiency.

An object of the present invention is to provide a series of techniques for producing isoprene from methanol or the like. Provided is a recombinant cell prepared by introducing a gene encoding isoprene synthase, into a host cell which is a methylotroph, wherein the gene is expressed in the host cell, and the recombinant cell is capable of producing isoprene from at least one C1 compound selected from the group consisting of methane, methanol, methylamine, formic acid, formaldehyde, and formamide. Preferably, it has at least one C1 carbon assimilating pathway selected from the group consisting of a serine pathway, a ribulose monophosphate pathway, and a xylulose monophosphate pathway as a fixing pathway of formaldehyde. Also provided is a method for producing isoprene using the recombinant cell.

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 engineered cells including ones having synthetic methylotrophy which include an NADH-dependent enzyme capable of converting G3P to 3PG (e.g., B. methanolicus gapN) and/or fructose-1,6-bisphosphatase, along with hexulose-6-phosphate synthase, 6-phospho-3-hexuloisomerase, a phosphoketolase, or a combination thereof. Engineered cells of the disclosure beneficially maintain adequate pool sizes of phosphorylated C3 and/or C4 compounds, and/or provide increased levels of NADPH. As such, the modifications allow for the generation of C6 compounds from C1 (e.g., a methanol feedstod) and C5 compounds, the regeneration of C5 compounds from C6 compounds by carbon rearrangement, and an improved balance between regeneration of C5 compounds and lower glycolysis. In turn, this allows the engineered microorganism to generate sufficient quantities of metabolic precursors (e.g., acetyl-CoA) which can be used in a bioproduct pathway, and the engineered cells can include further modifications to those pathway enzymes allowing for production of a desired bioproduct.

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.

An engineered pathway for carbon fixation without rubisco incorporates carbon dioxide into fructose-6-phosphate.