Sustainable Production of Plastic-Degrading Enzymes in Chlamydomonas pacifica

Sustainable production of plastic-degrading enzymes in Chlamydomonas pacifica Authors: Crisandra Jade Diaz1a, João Vitor Dutra Molino1a, Barbara Saucedo1, Kalisa Kang1, Évellin do Espirito Santo1,2, Marissa Tessman3, Michael D. Burkart3,4 Ryan Simkovsky1,4, Stephen Mayfield1,4 Affiliation: 1 - Division of Biological Sciences, University of California San Diego, La Jolla, California, United States of America. 2 - Department of Biochemical and Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil 3 - Department of Chemistry & Biochemistry, University of California San Diego, La Jolla, California, United States of America 4 - Algenesis Inc., 1238 Sea Village Dr., Cardiff, CA, United States of America a - Co-first authors 📁 code/ Description: Data analysis and figure-generation scripts in R; includes raw sensor data and curve-fitting files. File Description air_temp_sensor Raw air temperature data (used in Fig 3C). all sensors data Combined data from all pond sensors. light_sensor_DLI Daily Light Integral (DLI) calculations (used in Supp. Fig 2). light_sensor_MEAS Raw light intensity measurements (pre-processed). pH_temp_ponds Combined pH and temperature records from the raceway ponds. Pond_Impra Data from Impranil degradation assay (used in Fig 4C). production_curve, production_curve_pond Growth curve data from lab and pond experiments (used in Fig 2A and 3B). tpa_activity, TPA_standard Curve TPA release data from PET degradation assay and its calibration curve (used in Fig 2B). 📁 fig/Fig 1/ Figure 1 – Expression and activity of PHL7 in C. pacifica File Description 20240417 403 PHL7 PXL... Plate image showing halo formation (panel B). 20240509 Zymogram... (2 files) Zymogram gels showing polymer degradation by secreted PHL7 (panel C). 📁 fig/Fig 2/ Figure 2 – Growth and PET degradation File Description 20240429 Growth curve... Raw growth data comparing WT and PHL7 (panel A). 20240429 TPA activity... Raw absorbance data for TPA activity (panel B). growth_curve_plot Processed plot of growth over time (panel A). tpa_activity_prism_style Stylized bar graph of TPA absorbance (panel B). 📁 fig/Fig 3/ Figure 3 – Open pond cultivation File Description 20240314_ponds_growth Raw dry cell weight (DCW) data. all_data_merged_dataset Combined environmental and growth dataset. Light_sensor Light measurements over time (used in Fig 3C). pH_ponds_combined_results pH data across three ponds (used in Fig 3C). pond_growth_plot Processed DCW time-course figure (panel B). 📁 fig/Fig 4/ Figure 4 – Enzyme activity after pond cultivation File Description image2894, image2906 Impranil zymograms showing PHL7 activity from pond replicates (panels A & B). Pond_Impranil.xlsx Raw slope calculations and statistical analysis (panel C). 📁 Supplementary Figure 1/ Halo assay comparing wild-type and PHL7-expressing strains File Description PHL7_WT_plates_1 Photograph of plate showing halo formation by PHL7 strain and absence in WT. 📁 Supplementary Figure 2/ Daily light exposure (DLI) from two sensors File Description Light_sensor Averaged DLI values and variance between sensors. Used in bar chart. 📁 Supplementary Figure 3/ Vector map of plasmid pJP32PHL7 File Description pJP32PHL7 Map Annotated schematic of pJP32PHL7 plasmid. pJP32PHL7 (x2) Plasmid files in GenBank or SnapGene format (for cloning and annotation). 📁 vid 1/ Video 1 – Raceway pond operation File Description vid 1 Time-lapse or live video of 80 L open raceway pond with C. pacifica expressing PHL7. Demonstrates mixing, light exposure, and scale-up viability.