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 provides a breath test for assessing a metabolic phenotypeand/or assessing a disease state.

Described in this application are enzymes (e.g., cucurbitadienol synthases (CDS), UDP-glycosyltransferases (UGT), C11 hydroxylases, epoxide hydrolases (EPH), squalene epoxidases, and/or cytochrome P450 reductases), host cells expressing the enzymes, and methods of producing mogrol precursors, mogrol, and/or mogrosides using such host cells.

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.

Provided herein are compositions and methods for improved production of steviol glycosides in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding a Stevia rebaudiana kaurenoic acid hydroxylase. In some embodiments, the host cell further comprises one or more heterologous nucleotide sequence encoding further enzymes of a pathway capable of producing one or more steviol glycosides in the host cell. The compositions and methods described herein provide an efficient route for the heterologous production of steviol glycosides, including but not limited to, rebaudioside D and rebaudioside M.

Viral vectors to deliver a heterologous CYP4V2 gene to the retina, e.g., RPE cells of the retina, are provided herein to treat subjects with Bietti crystalline dystrophy.

Viral vectors to deliver a heterologous CYP4V2 gene to the retina, e.g., RPE cells of the retina, are provided herein to treat subjects with Bietti crystalline dystrophy.

The present invention provides transgenic animals (e.g., transgenic porcine animals), organs, tissues, and cells derived from the transgenic animals that are particularly useful for xenotransplantation therapies. In particular, the present invention provides transgenic porcine animals, as well as organs, tissues and cells derived from the transgenic porcine animals, which lack any expression of a functional alpha 1,3 galactosyltransferase (GTKO) gene and comprise at least six transgenes under the control of at least three promoters within a single multi-gene expression vector, and further comprise at least four additional genetic modifications. Also provided are methods of making the transgenic animals (e.g., transgenic porcine animals), and methods of using the transgenic animals, organs, tissues, and cells derived from the transgenic animals for xenotransplantation therapies and treating a disease or condition.

The invention relates to a long-chain dibasic acid with low content of monobasic acid impurity and a production method thereof, in particular to the preparation of a long-chain dibasic acid producing strain by means of directed evolution and homologous recombination, and to the production of a long-chain dibasic acid with low content of monobasic acid impurity by fermentation of said strain. The invention relates to a mutated CYP52A12 gene, homologous gene or variant thereof, which, relative to GenBank Accession Number AY230498 and taking the first base upstream of the start codon ATG as −1, comprises a mutation. The invention relates to a strain comprising said mutated CYP52A12 gene, homologous gene or variant thereof wherein when the strain is fermented to produce a long-chain dibasic acid, the content of monobasic acid impurity in the fermentation product is significantly reduced.

The invention is directed to a method of preparing a long chain dicarboxylic acid producing strain by using directed evolution and homologous recombination, a strain obtained by this method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use of the strain. In particular, the invention is directed to a method of preparing a long chain dicarboxylic acid producing strain by using directed evolution of CYP52A12 gene and homologous recombination, a strain obtained by this method that is capable of producing a long chain dicarboxylic acid under an acidic condition and the use of the strain.