DOI: 10.5281/zenodo.15411934
title: Sustainable Production of Plastic-Degrading Enzymes in Chlamydomonas pacifica

description: 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.