Reusable microsomal biocatalytic systems (bioreactors) constructed on carbon nanostructure modified electrodes are provided. The bioreactors comprise stable, biologically active immobilized enzymes such as human cytochromes P 450 (CYPs) and their redox partner proteins, e.g. CYP-NADPH (reduced nicotinamide adenine dinucleotide phosphate) reductases (CPR), on the carbon nanostructure surface. The immobilized enzymes may be present in liver microsomes, such as human liver microsomes (HLM) or as bactosomes, S9 fractions, etc. The bioreactors are used, for example, for synthesizing metabolites of interest from compounds such as drugs that are catabolized by the enzymes.

It is intended to provide a novel terpenoid derivative that exhibits anti-inflammatory action and a cytoprotective action by activating the Keap1/Nrf2/ARE signaling pathway. The present invention provides terpenoid derivative A represented by the following formula (I):

##STR00001##

It is intended to provide a novel terpenoid derivative that exhibits anti-inflammatory action and a cytoprotective action by activating the Keap1/Nrf2/ARE signaling pathway. The present invention provides terpenoid derivative A represented by the following formula (I):

##STR00001##

This invention relates to the production of 7-dehydrocholesterol, 25-hydroxy-7-dehydrocholesterol, and 25-hydroxy ergosterol in yeast such as Saccharomyces cerevisiae. It also relates to various enzymes catalyzing the reduction of the double bond at position 24 of lanosterol, dimethyl zymosterol, methyl zymosterol, zymosterol, cholesta-7, 24-dienol, or cholesta-5,7,24-trienol; or the hydroxylation at position 25 of ergosterol, 7-dehydrocholesterol, cholesta-8-enol, and cholesta-7-enol. It also relates to various nucleic acids encoding cholesterol C25-hydroxylases and sterol 24-reductases and their use to produce and hydroxylate 7-dehydrocholesterol or ergosterol. It also relates to the yeast strains so produced, and methods of making these sterols that include the steps of cultivaton a transformed yeast cell, and harvesting the resulting sterol(s).

This invention relates to the production of 7-dehydrocholesterol, 25-hydroxy-7-dehydrocholesterol, and 25-hydroxy ergosterol in yeast such as Saccharomyces cerevisiae. It also relates to various enzymes catalyzing the reduction of the double bond at position 24 of lanosterol, dimethyl zymosterol, methyl zymosterol, zymosterol, cholesta-7, 24-dienol, or cholesta-5,7,24-trienol; or the hydroxylation at position 25 of ergosterol, 7-dehydrocholesterol, cholesta-8-enol, and cholesta-7-enol. It also relates to various nucleic acids encoding cholesterol C25-hydroxylases and sterol 24-reductases and their use to produce and hydroxylate 7-dehydrocholesterol or ergosterol. It also relates to the yeast strains so produced, and methods of making these sterols that include the steps of cultivaton a transformed yeast cell, and harvesting the resulting sterol(s).

The present invention relates to an enzyme and method for the hydroxylation of a 7-deoxysteroid having the general formula (I) ##STR00001## at position 7 to a steroid having the general formula (II) ##STR00002##

The present invention relates to an enzyme and method for the hydroxylation of a 7-deoxysteroid having the general formula (I) ##STR00001## at position 7 to a steroid having the general formula (II) ##STR00002##

A chemoenzymatic method for synthesizing steroid HE3286 is provided, comprising: screening cytochrome P450 mutant LG-23 capable of catalyzing 7-hydroxylation of dehydroepiandrosterone; enzymatically converting dehydroepiandrosterone to 7-hydroxy-dehydroepiandrosterone using the P450 BM3 mutant enzyme; and chemically performing alkynylation at the C17th position carbonyl group to generate steroid HE3286. The steroid HE3286 synthesis method not only features simplified synthetic steps and high catalytic selectivity, but also offers mild reaction conditions, low cost, and environmentally friendly efficiency. This approach holds significant application value for advancing the development of steroid pharmaceuticals.

A chemoenzymatic method for synthesizing steroid HE3286 is provided, comprising: screening cytochrome P450 mutant LG-23 capable of catalyzing 7-hydroxylation of dehydroepiandrosterone; enzymatically converting dehydroepiandrosterone to 7-hydroxy-dehydroepiandrosterone using the P450 BM3 mutant enzyme; and chemically performing alkynylation at the C17th position carbonyl group to generate steroid HE3286. The steroid HE3286 synthesis method not only features simplified synthetic steps and high catalytic selectivity, but also offers mild reaction conditions, low cost, and environmentally friendly efficiency. This approach holds significant application value for advancing the development of steroid pharmaceuticals.

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.