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.

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.

Genetically-modified cell capable of producing UD CA, cholic acid, and/or another UDCA precursor comprising at least one heterologous polynucleotide encoding an enzyme involved in a metabolic pathway that converts sugar to UDCA, cholic acid, and/or another UDCA precursor. Method of making UDCA, cholic acid, and/or another UDCA precursor using such a cell. Use of UDCA or UDCA precursor produced using such a method for the manufacture of a medicament for the treatment of a disease or symptom of a disease. Medicament comprising UDCA or UDCA precursor made using such a method. Method of treating a disease or symptom of a disease comprising administering UDCA or a UDCA precursor made using such a method. Isolated nucleic acid encoding at least one enzyme involved in a metabolic pathway that converts sugar to UDCA, cholic acid, and/or another UDCA precursor. Vector comprising a nucleic acid encoding at least one enzyme involved in a metabolic pathway that converts sugar to UDCA, cholic acid and/or another UDCA precursor. Method of making a genetically-modified cell capable of synthesizing UDCA, cholic acid, and/or another UDCA precursor. Composition comprising UDCA or a UDCA precursor, a free acid or CoA thereof, or a pharmaceutically-acceptable derivative or prodrug thereof.

Genetically-modified cell capable of producing UD CA, cholic acid, and/or another UDCA precursor comprising at least one heterologous polynucleotide encoding an enzyme involved in a metabolic pathway that converts sugar to UDCA, cholic acid, and/or another UDCA precursor. Method of making UDCA, cholic acid, and/or another UDCA precursor using such a cell. Use of UDCA or UDCA precursor produced using such a method for the manufacture of a medicament for the treatment of a disease or symptom of a disease. Medicament comprising UDCA or UDCA precursor made using such a method. Method of treating a disease or symptom of a disease comprising administering UDCA or a UDCA precursor made using such a method. Isolated nucleic acid encoding at least one enzyme involved in a metabolic pathway that converts sugar to UDCA, cholic acid, and/or another UDCA precursor. Vector comprising a nucleic acid encoding at least one enzyme involved in a metabolic pathway that converts sugar to UDCA, cholic acid and/or another UDCA precursor. Method of making a genetically-modified cell capable of synthesizing UDCA, cholic acid, and/or another UDCA precursor. Composition comprising UDCA or a UDCA precursor, a free acid or CoA thereof, or a pharmaceutically-acceptable derivative or prodrug thereof.

A microbiological process is provided for the preparation of ursocholic acid, which includes the biotransformation of β-sitosterol in the presence of specific microorganisms.

A microbiological process is provided for the preparation of ursocholic acid, which includes the biotransformation of β-sitosterol in the presence of specific microorganisms.

The present invention relates to a recombinant yeast strain having sterol productivity, a preparation method therefor and a use thereof, and more specifically, to a recombinant yeast strain capable of producing cholesterol and cholesterol precursors in a high yield through the deletion of ERG5 and ERG6 genes and the introduction of DHCR24 and DHCR7 genes by codon-optimizing same in multiple or with a codon context method; and a production method therefor and a use thereof. In addition, disclosed are: a method for producing a recombinant yeast strain with increased production yields of cholesterol and cholesterol precursors by the additional introduction of gene tHMG1, ERG2, ERG3, ERG27, or UPC2-1 in the prepared recombinant yeast strain; and a use thereof

The present invention relates to a recombinant yeast strain having sterol productivity, a preparation method therefor and a use thereof, and more specifically, to a recombinant yeast strain capable of producing cholesterol and cholesterol precursors in a high yield through the deletion of ERG5 and ERG6 genes and the introduction of DHCR24 and DHCR7 genes by codon-optimizing same in multiple or with a codon context method; and a production method therefor and a use thereof. In addition, disclosed are: a method for producing a recombinant yeast strain with increased production yields of cholesterol and cholesterol precursors by the additional introduction of gene tHMG1, ERG2, ERG3, ERG27, or UPC2-1 in the prepared recombinant yeast strain; and a use thereof

The present disclosure discloses a method for producing 9α-hydroxy androstane-4-alkene-3,17-diketone by enzymatic conversion, and belongs to the fields of gene engineering and enzyme engineering. According to the present disclosure, oxidation subunit KshA, reduction subunit KshB and unknown active subunit KshC of 3-ketosteroid-9α-hydroxylase sourcing from Mycobacterium sp. Strain VKM Ac-1817D are successfully expressed in E. coli BL21, and KshC is identified as an oxidation subunit, the enzyme activity of which is far higher than that of KshA. BL21/pET-28a(+)-fdh constructed in the laboratory is used for expressing formate dehydrogenase (FDH), and by using crude enzyme liquid of KSH (KshB+KshC) and FDH engineering bacteria as a biocatalyst and a steroidal compound (AD) as a substrate, optimum reaction temperature is determined as 30° C. and optimum pH is determined as 7.0. In optimum conditions, AD is converted to produce a product 9-OH-AD, and within 20 hours, the output of 9-OH-AD is 4.7 g/L, and the molar conversion rate reaches 96.7%. According to the present disclosure, in production of 9-OH-AD, coupling of a 3-ketosteroid-9α-hydroxylase hydroxylation system and a coenzyme recycling system is realized, and the method has the advantages of being high in efficiency, low in cost, green, environmentally friendly and the like.

The present disclosure discloses a method for producing 9α-hydroxy androstane-4-alkene-3,17-diketone by enzymatic conversion, and belongs to the fields of gene engineering and enzyme engineering. According to the present disclosure, oxidation subunit KshA, reduction subunit KshB and unknown active subunit KshC of 3-ketosteroid-9α-hydroxylase sourcing from Mycobacterium sp. Strain VKM Ac-1817D are successfully expressed in E. coli BL21, and KshC is identified as an oxidation subunit, the enzyme activity of which is far higher than that of KshA. BL21/pET-28a(+)-fdh constructed in the laboratory is used for expressing formate dehydrogenase (FDH), and by using crude enzyme liquid of KSH (KshB+KshC) and FDH engineering bacteria as a biocatalyst and a steroidal compound (AD) as a substrate, optimum reaction temperature is determined as 30° C. and optimum pH is determined as 7.0. In optimum conditions, AD is converted to produce a product 9-OH-AD, and within 20 hours, the output of 9-OH-AD is 4.7 g/L, and the molar conversion rate reaches 96.7%. According to the present disclosure, in production of 9-OH-AD, coupling of a 3-ketosteroid-9α-hydroxylase hydroxylation system and a coenzyme recycling system is realized, and the method has the advantages of being high in efficiency, low in cost, green, environmentally friendly and the like.