Production of Recombinant Protein in the Chloroplast of Chlamydomonas Renhardtii: A Systems Biology Approach
Year: 2019
Type: article
Abstract: Research into recombinant protein production revealed some attractiveness with expression system based on microalgae and plant, for significant cost-savings and production of stable, active and correctly folded recombinant proteins. In this work, we undertook to study the growth of the microalgae, C. reinhardtii, for the chloroplast expression of recombinant E. coli β-glucuronidase (gus). We characterized the growth and recombinant gus protein yield in autotrophic, heterotrophic and mixotrophic conditions to understand limitations to growth and the protein yields. The maximum specific growth rate for cultures grown at different conditions: mixotrophic cultures (~0.05 h-1), autotrophic cultures (~0.03 h-1) and heterotrophic cultures (~0.025 h-1). The recombinant gus yields determined on the basis of percentage dry cell weight were: mixotrophic cultures (0.038%), autotrophic cultures (0.032%), heterotrophic cultures (0.026%). No detectable recombinant protein burden on the growth rate was observed for the production of recombinant gus in autotrophic and mixotrophic conditions, but significant reduction observed in the heterotrophic condition (~15%). The understanding of the factors that affected the growth of the cultures was used in designing suitable growth strategies in the heterotrophic condition that reduced the limitations and significantly increased growth ~3 fold. Additionally, flux balance analysis of the genome-scale metabolic re-constructed network of algae was used to provide insights and identify limitations by sub-optimal amino acid steady-state fluxes for gus production. Using the cycle of FBA modelling simulation for gus production, experimental verifications of predictions and FBA corrections, a strategy that significantly increased recombinant gus yield in heterotrophic cells by 2 fold.
Authors: Oluwafemi Davies, Malkhey Verma
Institutions: Central University of Punjab, University of Manchester
Language: English
Field: Energy
Domain: Physical Sciences
Open Access status: diamond
OpenAlex ID: W4235097196