Methods for Algal Protein Isolation and Proteome Analysis.

Eric P Knoshaug, Alida T Gerritsen, Calvin A Henard, Michael T Guarnieri

Methods in molecular biology (Clifton, N.J.) 2020

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Microalgae present promising feedstocks to produce renewable fuel and chemical intermediates, in part due to high storage carbon flux capacity to triacylglycerides or storage carbohydrates upon nutrient deprivation. However, the mechanism(s) governing deprivation-mediated carbon partitioning remain to be fully elucidated, limiting targeted strain engineering strategies in algal biocatalysts. Though genomic and transcriptomic analyses offer key insights into these mechanisms, active post-transcriptional regulatory mechanisms, ubiquitous in many microalgae, necessitate proteomic and post-translational (e.g., phospho- and nitroso-proteomic) analyses to more completely evaluate algal responsiveness to nutrient deprivation. Herein, we describe methods for isolating total algal protein and conducting proteomic, phosphoproteomic, and nitrosoproteomic analyses. We focus on methods deployed for the chlorophyte, Chlorella vulgaris, a model oleaginous alga with high flux to renewable fuel and chemical precursors.

Citation

Eric P Knoshaug, Alida T Gerritsen, Calvin A Henard, Michael T Guarnieri. Methods for Algal Protein Isolation and Proteome Analysis. Methods in molecular biology (Clifton, N.J.). 2020;2096:51-59