Abstract: Genetically engineered hosts and methods for their production and use in synthesizing hydrocarbons are provided.

Abstract: Nonnaturally occurring host cells altered to increase their ability to transfer genetic molecules into the host cells as compared to an unaltered host cell are provided. Also provided are methods for identifying endogenous loci of a host cell which inhibit transformation efficiency and/or electroporation of genetic molecules into the cell as well as methods for producing nonnaturally occurring host cells with enhanced transformation efficiency and/or the modified ability to allow for genomic integration of an exogenous DNA sequence via electroporation. Methods for producing biochemicals and products produced with the nonnaturally occurring host cells are also provided.

Abstract: Disclosed are methods for regulating biosynthesis of at least one of pimelic acid, 7-aminoheptanoic acid, 7-hydroxyheptanoic acid, heptamethylenediamine, 7-aminoheptanoland 1,7-heptanediol (C7 building blocks) using a pathway having a pimeloyl-ACP intermediate, the method including the step of downregulating the activity of BioF. Also disclosed are recombinant hosts by fermentation in which the above methods are performed. Further disclosed are recombinant hosts for producing pimeloyl-ACP, the recombinant host including a deletion of a bioF gene.

Abstract: This document describes biochemical pathways for producing a difunctional product having an odd number of carbon atoms in vitro or in a recombinant host, or salts or derivatives thereof, by forming two terminal functional groups selected from carboxyl, amine, formyl, and hydroxyl groups in an aliphatic carbon chain backbone having an odd number of carbon atoms synthesized from (i) acetyl-CoA and propanedioyl-CoA via one or more cycles of methyl ester shielded carbon chain elongation or (ii) propanedioyl-[acp] via one or more cycles of methyl ester shielded carbon chain elongation. The biochemical pathways and metabolic engineering and cultivation strategies described herein rely on enzymes or homologs accepting methyl ester shielded aliphatic carbon chain backbones and maintaining the methyl ester shield for at least one further enzymatic step following one or more cycles of methyl ester shielded carbon chain elongation.

Abstract: Methods and compositions for synthesizing dienes and derivative thereof, such as isoprene, in Cupriavidus necator are provided.

Abstract: Nonnaturally occurring host cells altered to increase their ability to transfer genetic molecules into the host cells as compared to an unaltered host cell are provided. Also provided are methods for identifying endogenous loci of a host cell which inhibit transformation efficiency and/or electroporation of genetic molecules into the cell as well as methods for producing nonnaturally occurring host cells with enhanced transformation efficiency and/or the modified ability to allow for genomic integration of an exogenous DNA sequence via electroporation. Methods for producing biochemicals and products produced with the nonnaturally occurring host cells are also provided. In particular, a host Cupriavidus necator cell is provided in which a chromosomal gene encoding a restriction endonuclease has been disrupted, leading to an increase in transformation efficiency.

Abstract: Methods and materials for the production of beta hydroxy acids, such as 3-hydroxypropanoic acid (3-HP) and/or derivatives thereof and/or compounds related thereto, are provided. Also provided are products produced in accordance with these methods and materials.

Abstract: Methods and compositions for synthesizing dienes and derivative thereof, such as isoprene, in Cupriavidus necator are provided.

Abstract: Genetically engineered hosts and methods for their production and use in synthesizing hydrocarbons are provided.

Abstract: Nonnaturally occurring host cells altered to increase their ability to transfer genetic molecules into the host cells as compared to an unaltered host cell are provided. Also provided are methods for identifying endogenous loci of a host cell which inhibit transformation efficiency and/or electroporation of genetic molecules into the cell as well as methods for producing nonnaturally occurring host cells with enhanced transformation efficiency and/or the modified ability to allow for genomic integration of an exogenous DNA sequence via electroporation. Methods for producing biochemicals and products produced with the nonnaturally occurring host cells are also provided.

Abstract: Methods and compositions for synthesizing dienes and derivative thereof, such as isoprene, in Cupriavidus necator are provided.

Abstract: This document describes biochemical pathways for producing a difunctional product having an odd number of carbon atoms in vitro or in a recombinant host, or salts or derivatives thereof, by forming two terminal functional groups selected from carboxyl, amine, formyl, and hydroxyl groups in an aliphatic carbon chain backbone having an odd number of carbon atoms synthesized from (i) acetyl-CoA and propanedioyl-CoA via one or more cycles of methyl ester shielded carbon chain elongation or (ii) propanedioyl-[acp] via one or more cycles of methyl ester shielded carbon chain elongation. The biochemical pathways and metabolic engineering and cultivation strategies described herein rely on enzymes or homologs accepting methyl ester shielded aliphatic carbon chain backbones and maintaining the methyl ester shield for at least one further enzymatic step following one or more cycles of methyl ester shielded carbon chain elongation.

Abstract: Disclosed are methods for regulating biosynthesis of at least one of pimelic acid, 7-aminoheptanoic acid, 7-hydroxyheptanoic acid, heptamethylenediamine, 7-aminohelptanol and 1,7-heptanediol (C7 building blocks) using a pathway having a pimeloyl-ACP intermediate, the method including the step of downregulating the activity of BioF. Also disclosed are recombinant hosts by fermentation in which the above methods are performed. Further disclosed are recombinant hosts for producing pimeloyl-ACP, the recombinant host including a deletion of a bioF gene.

Abstract: The invention relates to a method of double crossover homologous recombination in a host Clostridia cell comprising: a first homologous recombination event between a donor DNA molecule and DNA of the host cell to form a product of the first recombination event in the host cell, wherein the donor DNA molecule comprises a codA gene and at least two homology arms; and a second recombination event within the product of the first homologous recombination event, thereby to form a product of the second homologous recombination event in the host cell which is selectable by the loss of the codA gene; and a related vector and altered host cell.