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

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.
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
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12079/51629
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 11
social impact