Labdane diterpenes (LDs), and especially sclareol, are important feedstocks for the pharmaceutical and cosmetic industries, and therefore several lines of research have led to their heterologous production in non-photosynthetic microbes and higher plants. The potential of microalgae as bioreactors of natural products has been established for a variety of bioactive metabolites, including terpenes. In this work, a codon optimized sequence encoding a key plant labdane-type diterpene (LD) cyclase, copal-8-ol diphosphate synthase from Cistus creticus (CcCLS), was introduced into the chloroplast genome of Chlamydomonas reinhardtii. Of 49 transplastomic algal lines, 12 produced variable amounts of four LD compounds, namely ent-manoyl oxide, sclareol, labda-13-ene-8α,15-diol and ent-13-epi-manoyl oxide. The total LD concentrations measured in the transplastomic lines reached 1.172 ± 0.05 μg/mg cell DW for the highest overall producer, while the highest yield for sclareol was 0.038 ± 0.001 μg/mg cell DW. Thus, transplastomic expression of a key plant labdane diterpene cyclase in the C. reinhardtii chloroplast genome enabled the production of important plant-specific LD compounds.

Heterologous production of labdane-type diterpenes in the green alga Chlamydomonas reinhardtii

Diretto G.;Demurtas O. C.;Mini P.;Ferrante P.;Giuliano G.;
2019-01-01

Abstract

Labdane diterpenes (LDs), and especially sclareol, are important feedstocks for the pharmaceutical and cosmetic industries, and therefore several lines of research have led to their heterologous production in non-photosynthetic microbes and higher plants. The potential of microalgae as bioreactors of natural products has been established for a variety of bioactive metabolites, including terpenes. In this work, a codon optimized sequence encoding a key plant labdane-type diterpene (LD) cyclase, copal-8-ol diphosphate synthase from Cistus creticus (CcCLS), was introduced into the chloroplast genome of Chlamydomonas reinhardtii. Of 49 transplastomic algal lines, 12 produced variable amounts of four LD compounds, namely ent-manoyl oxide, sclareol, labda-13-ene-8α,15-diol and ent-13-epi-manoyl oxide. The total LD concentrations measured in the transplastomic lines reached 1.172 ± 0.05 μg/mg cell DW for the highest overall producer, while the highest yield for sclareol was 0.038 ± 0.001 μg/mg cell DW. Thus, transplastomic expression of a key plant labdane diterpene cyclase in the C. reinhardtii chloroplast genome enabled the production of important plant-specific LD compounds.
2019
Chlamydomonas reinhardtii P.A.Dangeard (Chlamydomonadaceae); Cistus creticus L. (Cistaceae); Copal-8-ol diphosphate synthase; Ent-13-epi-manoyl oxide; Ent-manoyl oxide; Heterologous production; Labda-13-ene-8α,15-diol; Labda-14-ene-8,13-diol (sclareol); Labdane diterpenes; Microalgae; Alkyl and Aryl Transferases; Chlamydomonas reinhardtii; Chloroplasts; Cistus; Diterpenes; Plant Proteins; Transformation, Genetic; Genetic Engineering
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12079/51629
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