The subject of the present invention is a whole-cell catalysis process for converting substrates of cytochrome P450 monooxygenases of eukaryotic origin into valuable biotechnological products. The subject of the present invention is also microorganisms genetically engineered to achieve those biotransformations with high rates and processes to prepare these microorganism strains.

Hybrid proteins with cytochrome P450 activity and which are encapsulated in a nanocapsid (nanoparticles charged with cytochrome P450 activity) are designed and synthesized, these hybrid proteins being immunologically inert and recognized by breast cancer cells.

The present invention relates to novel process for the production of ketoisophorone via biocatalytic conversion of isophorone, in particular a one-pot biocatalytic system for conversion of ?-isophorone in a two-step oxidation process, with a first oxidation being catalyzed by a heme containing oxidoreductase such as a cytochrome P450 monooxygenase followed by another oxidation which can be either a chemical reaction or a biocatalytic reaction, in particular wherein the oxidation is catalyzed by an NAD(P) or NADP(H)-dependent oxidoreductase. The invention further provides polypeptides and nucleic acid sequences coding for cytochrome P450 monooxygenase with modified (higher) substrate selectivity, total turnover numbers and/or (re)activity compared to the wild-type enzyme. Ketoisophorone is useful as building block in the synthesis of vitamins and carotenoids.

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.

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.

Provided are a cytochrome P450 enzyme mutant and an application thereof. The enzyme activity and anti-Markov oxidation selectivity of P450 derived from a wild-type strain of Bacillus megaterium undergo protein modification by means of directed evolution, thus improving enzyme activity and selectivity, and developing a series of P450 enzyme mutants that may be used for industrial production.