Combinatorial engineering for photoautotrophic production of recombinant products from the green microalga Chlamydomonas reinhardtii
KAUST DepartmentBioengineering Program, Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Kingdom of Saudi Arabia
Marine Science Program
Biological and Environmental Science and Engineering (BESE) Division
Permanent link to this recordhttp://hdl.handle.net/10754/677956
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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 and transgene expression engineering 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 use combinatorial chloroplast and nuclear genome engineering to complement the C. reinhardtii strain UVM4 with publicly available genetic tools to enable the use of inorganic phosphite and nitrate as a sole the source phosphorous and nitrogen, respectively. We present recipes to create phosphite-buffered media solutions that enable high cell density algal cultivation. We then combine 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 under 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.
CitationAbdallah, M. N., Wellman, G. B., Overmans, S., & Lauersen, K. J. (2022). Combinatorial engineering for photoautotrophic production of recombinant products from the green microalga Chlamydomonas reinhardtii. https://doi.org/10.1101/2022.02.28.482248
SponsorsSubcloning 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 Lauersen (KAUST). The authors are grateful to Saul Purton 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. The research reported in this publication was supported by baseline funding from KAUST to KL.
PublisherCold Spring Harbor Laboratory