The present invention relates to an enzme and method for the hydroxylation of a 7-deoxysteroid having the general formula (I)

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at position 7 to a steroid having the general formula (II)

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The present invention relates to an enzme 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##

In certain aspects, the present invention provides methods for producing terminally hydroxylated alkenes and alkynes by contacting an unsaturated or saturated hydrocarbon substrate with a hydroxylase enzyme. Exemplary terminal hydroxylases useful for carrying out the methods of the invention exhibit strong selectivity towards one terminal carbon of a hydrocarbon substrate and include, but are not limited to, non-heme diiron alkane monooxygenases, cytochromes P450 (e.g., cytochromes P450 of the CYP52 and CYP153 family), as well as long chain alkane hydroxylases. In some embodiments, the terminally hydroxylated alkene or alkyne is further converted to a terminal alkenal. In certain embodiments, terminally hydroxylated alkenes and alkynes are useful as insect pheromones which modify insect behavior. In other embodiments, terminally hydroxylated alkenes and alkynes are useful intermediates for producing pheromones via acetylation or oxidation of the alcohol moiety.

The invention relates to novel 7-hydroxysteroid dehydrogenases which are obtainable from bacteria of the genus Collinsella, especially of the strain Collinsella aerofaciens, to the sequences encoding said enzymes, to methods for producing said enzymes and to their use in the enzymatic conversion of cholic acid compounds, and especially in the production of ursodeoxycholic acid (UDCS). The invention also relates to novel methods for the synthesis UDCS.

The invention relates to novel 7-hydroxysteroid dehydrogenases which are obtainable from bacteria of the genus Collinsella, especially of the strain Collinsella aerofaciens, to the sequences encoding said enzymes, to methods for producing said enzymes and to their use in the enzymatic conversion of cholic acid compounds, and especially in the production of ursodeoxycholic acid (UDCS). The invention also relates to novel methods for the synthesis UDCS.

The present inventors have found that the content ratios of isoalloLCA, 3-oxoLCA, alloLCA, and 3-oxoalloLCA in the feces of centenarians are higher than those of younger ones, and have also identified gut microbiomes peculiar to centenarians involved in the production of these bile acids. Furthermore, it has been found that these bile acids, enzymes involved in the production thereof, and bacteria producing the enzymes reduce the risk of infection with pathogens, prostate cancer, and the like, and are involved in longevity.

The present inventors have found that the content ratios of isoalloLCA, 3-oxoLCA, alloLCA, and 3-oxoalloLCA in the feces of centenarians are higher than those of younger ones, and have also identified gut microbiomes peculiar to centenarians involved in the production of these bile acids. Furthermore, it has been found that these bile acids, enzymes involved in the production thereof, and bacteria producing the enzymes reduce the risk of infection with pathogens, prostate cancer, and the like, and are involved in longevity.

The disclosure provides compositions and methods related to engineered microbial cells, enzymes, and methods for producing dammarenediol, as well as compounds derived from dammarenediol. Microbial host cells are engineered to express a heterologous biosynthetic pathway that produces dammarenediol, or a derivative thereof. The host cell can optionally express a heterologous uridine diphosphate-dependent glycosyltransferase (UGT) enzyme producing natural or non-natural glycosylated forms of dammarenediol, protopanaxadiol or protopanaxatriol.

The disclosure provides compositions and methods related to engineered microbial cells, enzymes, and methods for producing dammarenediol, as well as compounds derived from dammarenediol. Microbial host cells are engineered to express a heterologous biosynthetic pathway that produces dammarenediol, or a derivative thereof. The host cell can optionally express a heterologous uridine diphosphate-dependent glycosyltransferase (UGT) enzyme producing natural or non-natural glycosylated forms of dammarenediol, protopanaxadiol or protopanaxatriol.

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.