Biosynthesis of retinoids
AssigneeDsm IP Assets B.V.
InventorsNathalie Balch, Paul Blomquist, Reed Doten, Peter Houston, Ethan Lam, Jenna McMahon, Joshua Trueheart, Celine Viarouge
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Abstract
The present invention is related to a novel enzymatic process for production of retinoids via a multi-step process, which process includes the use of heterologous enzymes having activity in a carotene-producing host cell, particularly wherein such process results in high percentage of retinoids, in trans-isoform.
Claims17
A process for production of trans-retinyl acetate comprising cultivating a retinoid-producing fungal host cell in an aqueous medium under suitable culture conditions, wherein
- (a) the host cell expresses
- (i) a heterologous stereo-selective beta-carotene oxidizing enzyme (BCO) capable of catalyzing the conversion of beta-carotene into a retinal mix comprising cis- and trans-retinal,
- (ii) a heterologous retinol dehydrogenase (RDH) enzyme [EC 1.1.1.105] capable of converting retinal into retinol, and
- (iii) a heterologous fungal acetyl transferase 1 enzyme (ATF1) [EC 2.3.1.84] capable of catalysing the conversion of trans-retinol to a retinyl acetate, and
- (b) the host cell is capable of
- (i) producing a retinal mix comprising cis- and trans-retinal, wherein the percentage of trans-retinal in the mix is at least 90%,
- (ii) converting the retinal mix into retinol with a total conversion of at least 90% towards generation of retinol, and
- (iii) converting the retinol into retinyl acetate, wherein at least 40% of the total amount of retinoids produced by said host cell is retinyl acetate.
A process for production of vitamin A comprising the steps of:
- (a) introducing a nucleic acid molecule encoding stereo-selective BCO, acetyl transferase [EC 2.3.1.84], and retinol dehydrogenase [EC 1.1.1.105] enzymes into a suitable fungal host cell;
- and
- (b) expressing said introduced enzymes to enable enzymatic conversion of beta-carotene into retinal comprising at least a percentage of 90% in trans isoform based on the total amount of retinoids produced by the host cell, to enable enzymatic conversion of said retinal into retinol comprising a percentage of at least 90% towards generation of retinol and to enable enzymatic conversion of retinol into retinyl acetate, wherein at least 40% of the total amount of retinoids produced by said host cell is retinyl acetate, said retinyl acetate comprising at least a percentage of 90% in trans isoform.
Dependencies are inferred from the claim text.