The present invention relates to the production of chemical species in bacterial host cells. Particularly, the present invention provides for the production of chemical species in Escherichia coli (E. coli) host cells that functionally express engineered P450 enzymes.

The subject of the present invention is a process for preparing a genetically modified yeast by multicopy integration of at least four expression cassettes, allowing the production of a molecule of interest at high titre. The subject of the present invention is also yeasts transformed according to said process, and the use thereof for producing hydrocortisone.

This invention relates generally to methods and compositions for producing a sesquiterpene alcohol comprising contacting a sesquiterpene with a P450 polypeptide with monooxygenase activity.

The subject of the present invention is a process for preparing a genetically modified yeast by multicopy integration of at least four expression cassettes, allowing the production of a molecule of interest at high titer. The subject of the present invention is also yeasts transformed according to said process, and the use thereof for producing hydrocortisone.

This invention relates generally to methods and compositions for producing a sesquiterpene alcohol comprising contacting a sesquiterpene with a p450 polypeptide with monooxygenase activity.

The present invention relates to novel monooxygenases which are useful in the hydroxylation of aromatic hydrocarbons. They are particularly useful for the production of 1-naththol and 7-hydroxycoumarin from naphthol and 7-Ethoxycoumarin, respectively.

The present invention pertains to an isolated P450 enzyme comprising or consisting of an amino acid sequence at least 80% identical to SEQ ID NO: 1, wherein said sequence comprises a threonine at position corresponding to position 225 and/or an aspartic acid mutation at position corresponding to position 289. The invention also concerns an isolated nucleic acid comprising a sequence encoding said enzyme, a vector comprising said nucleic acid, and a host cell containing said nucleic acid or said vector. Methods for preparing said enzyme and methods for producing steroid hormone precursors using the enzyme or the host cells featured in the invention are also provided.

Described herein are CB5 polypeptides for the biosynthesis of oxygenated hydrocarbons in modified host cells.

The present invention relates to a fusion protein, to a nucleic acid coding for the protein, to a vector comprising the nucleic acid, to a host cell comprising the nucleic acid and/or vector, to a process for producing a fusion protein and to a process for bioconverting a substrate. The fusion protein of the present invention successively comprises (i) at least one polypeptide for targeting, and anchoring to, the bacterial membrane, (ii) at least one polypeptide corresponding to the hydrophilic domain of a plant P450 cytochrome, (iii) at least one binding polypeptide comprising at least 47 amino acids, and (iv) at least one polypeptide corresponding to the hydrophilic domain of a plant NADPH-dependent cytochrome P450 reductase.