A gene editing system of Candida viswanathii includes a Candida viswanathii, a first gene editing fragment and a second gene editing fragment. The first gene editing fragment successively includes a first homology arm and a screening gene. The second gene editing fragment is connected to a C-terminus of the first gene editing fragment and includes a second homology arm, a Cas9 expression cassette and a sgRNA cassette. The Cas9 expression cassette successively includes a Cas9 promoter, a Cas9 gene and three nuclear localization sequences. The sgRNA cassette successively includes a sgRNA promoter, a first ribozyme, a targeting sequence, a scaffold and a second ribozyme. The first gene editing fragment and the second gene editing fragment are constructed as a linear fragment for gene editing of a chromosome of the Candida viswanathii.

Generally, the present invention relates to the field of steroid hydroxylation. More specifically, the present invention relates to a method for the 21-hydroxylation of steroids in cells. It also relates to cells expressing a steroid 21-hydroxylating enzyme or steroid 21-hydroxylase, expression vectors comprising a nucleic acid encoding for a steroid 21-hydroxylase and a kit for carrying out the method for the 21-hydroxylation of steroids in cells.

The invention relates to the directed evolution of CYP52A12 gene and the use thereof for the production of a dicarboxylic acid. In particular, it relates to a method of preparing a long chain dicarboxylic acid producing strain by using directed evolution and homologous recombination, a strain obtained by the method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use thereof. In particular, the invention relates to a method of preparing a long chain dicarboxylic acid producing strain by directed evolution of CYP52A12 gene and homologous recombination, a strain obtained by the method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use thereof. By directed evolution of CYP52A12 gene, one strain which has a base mutation at the promoter region of said gene and is capable of producing a long chain dicarboxylic acid under an acidic condition in a shortened fermentation time is screened out in the invention.

The present invention provides a method of producing ()-rotundone from -guaiene. The method includes the steps of: (1) allowing a cytochrome P450 protein to act on -guaiene, which cytochrome P450 belongs to the CYP152 family and is capable of oxidizing the methylene group at position 3 of -guaiene to the carbonyl group; and/or (2) allowing a cytochrome P450 protein to act on -guaiene in the presence of an electron transfer protein capable of transferring electrons to the cytochrome P450 protein, which cytochrome P450 belongs to the CYP152, CYP106, or CYP107 family and is capable of oxidizing the methylene group at position 3 of -guaiene to the carbonyl group.

Generally, the present invention relates to the field of steroid hydroxylation. More specifically, the present invention relates to a method for the 21-hydroxylation of steroids in cells. It also relates to cells expressing a steroid 21-hydroxylating enzyme or steroid 21-hydroxylase, expression vectors comprising a nucleic acid encoding for a steroid 21-hydroxylase and a kit for carrying out the method for the 21-hydroxylation of steroids in cells.

The present invention relates to a recombinant microorganism which is modified to be capable of producing diosmin and hesperidin and to the use thereof for producing diosmin and/or hesperidin.

The present invention relates to a method of preparing a steroid comprising the step of converting a 7-deoxysteroid with a cytochrome P450 enzyme or a functional variant thereof in the presence of at least one redox partner system and a system for regenerating the redox partner system.

Disclosed herein are recombinant adeno-associated viral vectors expressing 21-hydroxy lase (21OH) protein and related uses for treating 21OH deficiency.

The present invention relates a variant polypeptide having kaurenoic acid 13-hydroxylase activity, which variant polypeptide comprises an amino acid sequence which, when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1, comprises at least one substitution of an amino acid residue corresponding to any of amino acids 72, 85, 108, 127, 129, 141, 172, 195, 196, 197, 199, 226, 236, 291, 302, 361 or 464, said positions being defined with reference to SEQ ID NO: 1 and wherein the variant has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A variant polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

Embodiments described herein relate to suppressing the immune response locally within tissue transplants and certain conditions improperly affecting the immune system using optogenetically controlled cells. More specifically, embodiments described herein provide for localized immunosuppression surrounding tissue transplants and illness locations as an alternative to systemically suppressing a patient's entire immune system. Methods include implantation of optogenetically modified immunosuppressive cells that are configured to alter their biological activity to enhance their immunosuppressive activity in response to exposure of wavelengths of light in the red and near-infrared window spectral region (620-900 nm).