Engineering Nannochloropsis oceanica for the production of diterpenoid compounds

Du, Zhi‐Yan; Bhat, Wajid W.; Poliner, Eric; Johnson, Sean; Bertucci, Conor; Farre, Eva; Hamberger, Bjoern

doi:10.1002/mlf2.12097

Title: Engineering Nannochloropsis oceanica for the production of diterpenoid compounds

Journal Article · 25 December 2023 · mLife

DOI: https://doi.org/10.1002/mlf2.12097 · OSTI ID:2263331

^[1]; Bhat, Wajid W. ^[2]; Poliner, Eric ^[3]; Johnson, Sean ^[2]; Bertucci, Conor ^[2]; Farre, Eva ^[3]; Hamberger, Bjoern ^[2]

Department of Molecular Biosciences and Bioengineering University of Hawaii at Manoa Honolulu Hawaii USA
Department of Biochemistry and Molecular Biology Michigan State University East Lansing Michigan USA
Department of Plant Biology Michigan State University East Lansing Michigan USA

Abstract Photosynthetic microalgae like Nannochloropsis hold enormous potential as sustainable, light‐driven biofactories for the production of high‐value natural products such as terpenoids. Nannochloropsis oceanica is distinguished as a particularly robust host with extensive genomic and transgenic resources available. Its capacity to grow in wastewater, brackish, and sea waters, coupled with advances in microalgal metabolic engineering, genome editing, and synthetic biology, provides an excellent opportunity. In the present work, we demonstrate how N. oceanica can be engineered to produce the diterpene casbene—an important intermediate in the biosynthesis of pharmacologically relevant macrocyclic diterpenoids. Casbene accumulated after stably expressing and targeting the casbene synthase from Daphne genkwa (DgTPS1) to the algal chloroplast. The engineered strains yielded production titers of up to 0.12 mg g ⁻¹ total dry cell weight (DCW) casbene. Heterologous overexpression and chloroplast targeting of two upstream rate‐limiting enzymes in the 2‐C‐methyl‐ d ‐erythritol 4‐phosphate pathway, Coleus forskohlii 1‐deoxy‐ d ‐xylulose‐5‐phosphate synthase and geranylgeranyl diphosphate synthase genes, further enhanced the yield of casbene to a titer up to 1.80 mg g ⁻¹ DCW. The results presented here form a basis for further development and production of complex plant diterpenoids in microalgae.

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Sponsoring Organization:: USDOE

Grant/Contract Number:: SC0018409

OSTI ID:: 2263331

Journal Information:: mLife, Journal Name: mLife Journal Issue: 4 Vol. 2; ISSN 2097-1699

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: China

Language:: English

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