Combinatorial Engineering Enables Photoautotrophic Growth in High Cell Density Phosphite-Buffered Media to Support Engineered Chlamydomonas reinhardtii Bio-Production Concepts
KAUST DepartmentBioengineering Program
Biological and Environmental Science and Engineering (BESE) Division
Marine Science Program
Red Sea Research Center (RSRC)
Permanent link to this recordhttp://hdl.handle.net/10754/677872
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AbstractChlamydomonas reinhardtii has emerged as a powerful green cell factory for metabolic engineering of sustainable products created from the photosynthetic lifestyle of this microalga. Advances in nuclear genome modification and transgene expression are allowing robust engineering strategies to be demonstrated in this host. However, commonly used lab strains are not equipped with features to enable their broader implementation in non-sterile conditions and high-cell density concepts. Here, we used combinatorial chloroplast and nuclear genome engineering to augment the metabolism of the C. reinhardtii strain UVM4 with publicly available genetic tools to enable the use of inorganic phosphite and nitrate as sole sources of phosphorous and nitrogen, respectively. We present recipes to create phosphite-buffered media solutions that enable high cell density algal cultivation. We then combined previously reported engineering strategies to produce the heterologous sesquiterpenoid patchoulol to high titers from our engineered green cell factories and show these products are possible to produce in non-sterile conditions. Our work presents a straightforward means to generate C. reinhardtii strains for broader application in bio-processes for the sustainable generation of products from green microalgae.
CitationAbdallah, M. N., Wellman, G. B., Overmans, S., & Lauersen, K. J. (2022). Combinatorial Engineering Enables Photoautotrophic Growth in High Cell Density Phosphite-Buffered Media to Support Engineered Chlamydomonas reinhardtii Bio-Production Concepts. Frontiers in Microbiology, 13. https://doi.org/10.3389/fmicb.2022.885840
SponsorsFunding for this work was supported by the King Abdullah University of Science and Technology baseline research fund awarded to KL.
Subcloning of PcPS plasmid 1X was performed in the lab of Prof. Dr. Olaf Kruse by Dr. Julian Wichmann and Dr. Thomas Baier as part of an Institute for Innovation Transfer (IIT), Universität Bielefeld project funded by KL (KAUST). The authors are grateful to Saul Purton (UCL) for providing plasmid pPO3 and Prof. Dr. Ralph Bock for graciously providing strain UVM4 through MTA between the Max Planck Institute of Molecular Physiology and KAUST. We would like to express thanks to SSB group members for cooperation and collaboration during this project.
PublisherFrontiers Media SA
JournalFrontiers in Microbiology
Except where otherwise noted, this item's license is described as Archived with thanks to Frontiers in Microbiology under a Creative Commons license, details at: https://creativecommons.org/licenses/by/4.0/