Investigating factors that influence microalgae production platforms for diterpenoids in laboratory and outdoor conditions
type: CreativeWork
description: Photosynthetic microalgae offer a biotechnological solution to current climate concerns and the growing demand for food, bioproducts and bulk chemicals. As naturally efficient producers of carbohydrates, lipids, proteins and pigments, microalgae are a sustainable platform in which to realise the demand for high-value compounds. Moreover, they are amenable to metabolic engineering to produce other such molecules, including those with medicinal or therapeutic potential. The challenge however is to find solutions to the current bottlenecks, a major one of which is the challenge of microalgal scale-up. This project uses strains of the green microalga *Chlamydomonas reinhardtii*, UVM4 which is cell-wall deficient and RSW2 that has an intact cell wall. Both strains had previously been engineered to produce the diterpenoid, casbene, by introduction of the *Jatropha curcas* gene encoding casbene synthase. Two constructs were used, PM50, where the casbene synthase gene is under the control of the HSP70/RBCS2 promoter and includes a chloroplast transit peptide, and PM51, which in addition includes a thiamine dependent riboswitch (CrTHI4_4N). Casbene-producing *C. reinhardtii* strains were characterised under standard laboratory conditions of 25ºC and 90 μmol s-1 m-2 in Tris-Acetate Phosphate media (TAP) and found to reliably produce casbene when measured by gas-chromatography mass-spectrometry. Casbene production in strains ranged from 0.064 ± 0.074 mg l-1 to 16.7 ± 0.66 mg l-1 with RSW2::PM50#B08 being the best. The results of this work provided a reference against which to compare diterpenoid production in outdoor conditions. Casbene-producing strains RSW2::PM50#B08, UVM4::PM50#B12 and UVM4::PM51#E09 were taken forward for experiments in the outdoor facility located in the Botanic Garden at the University of Cambridge. Strains were inoculated in 10 l hanging bags, and the impact of environmental conditions during Summer and Autumn, the walled versus cell wall deficient phen…
abstract: Photosynthetic microalgae offer a biotechnological solution to current climate concerns and the growing demand for food, bioproducts and bulk chemicals. As naturally efficient producers of carbohydrates, lipids, proteins and pigments, microalgae are a sustainable platform in which to realise the demand for high-value compounds. Moreover, they are amenable to metabolic engineering to produce other such molecules, including those with medicinal or therapeutic potential. The challenge however is to find solutions to the current bottlenecks, a major one of which is the challenge of microalgal scale-up. This project uses strains of the green microalga *Chlamydomonas reinhardtii*, UVM4 which is cell-wall deficient and RSW2 that has an intact cell wall. Both strains had previously been engineered to produce the diterpenoid, casbene, by introduction of the *Jatropha curcas* gene encoding casbene synthase. Two constructs were used, PM50, where the casbene synthase gene is under the control of the HSP70/RBCS2 promoter and includes a chloroplast transit peptide, and PM51, which in addition includes a thiamine dependent riboswitch (CrTHI4_4N). Casbene-producing *C. reinhardtii* strains were characterised under standard laboratory conditions of 25ºC and 90 μmol s-1 m-2 in Tris-Acetate Phosphate media (TAP) and found to reliably produce casbene when measured by gas-chromatography mass-spectrometry. Casbene production in strains ranged from 0.064 ± 0.074 mg l-1 to 16.7 ± 0.66 mg l-1 with RSW2::PM50#B08 being the best. The results of this work provided a reference against which to compare diterpenoid production in outdoor conditions. Casbene-producing strains RSW2::PM50#B08, UVM4::PM50#B12 and UVM4::PM51#E09 were taken forward for experiments in the outdoor facility located in the Botanic Garden at the University of Cambridge. Strains were inoculated in 10 l hanging bags, and the impact of environmental conditions during Summer and Autumn, the walled versus cell wall deficient phen…
author: Tran, Nhan-An
language: eng
keywords: biotechnology, diterpenoid, microalgae, scale-up, synthetic biology
license: https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/60d510c8-f519-49b9-8c8a-ae4256ea9eb8/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2023-01-01
date modified: 2026-07-22T22:24:30Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.99765