DOI: 10.5281/zenodo.13981201
title: Dataset_Efficient secretion of a plastic degrading enzyme from the green algae Chlamydomonas reinhardtii

description: Manuscript Information:





Title: Efficient secretion of a plastic degrading enzyme from the green algae Chlamydomonas reinhardtii




Authors: João Vitor Dutra Molino, Barbara Saucedo, Kalisa Kang, Chloe Walsh, Crisandra Jade Diaz, Marissa Tessman, Ryan Simkovsky, Stephen Mayfield




Affiliation:






Division of Biological Sciences, University of California San Diego, La Jolla, California, United States of America




Algenesis Inc., 1238 Sea Village Dr., Cardiff, CA, United States of America





Correspondence: Stephen P. Mayfield (smayfield@ucsd.edu)




Keywords: Microalgae, Chlamydomonas reinhardtii, Environmental Biotechnology, Climate-Neutral Economy, Sustainable Plastic Recycling, PETase, secretion



Data Overview:

This dataset supports the manuscript, specifically regarding the efficient secretion of the plastic-degrading enzyme PHL7 from Chlamydomonas reinhardtii expressing the pJP32PHL7 vector. The data include various assays, results, and visual documentation of enzymatic activity, protein sequencing, colony growth, and time-lapse imagery.

CSV Files:





FDA_Screening_colonies_05_impranil.csv: Data from an FDA (fluorescein diacetate) enzymatic assay performed on colonies from plates with 0.5% Impranil®. This assay tracks enzyme activity related to plastic degradation.




FDA_Screening_colonies_075_impranil.csv: Similar FDA assay data, but from colonies on plates containing 0.75% Impranil®.




Impranil_Agarose_Abs_spectra.csv: Absorbance spectra data showing the activity of PHL7 in degrading Impranil® within an agarose medium (0.2% agarose, 0.25% Impranil®).




Impranil_Screening_colonies_05_impranil.csv: Data from enzymatic reactions, specifically tracking the degradation of PET plastic beads by PHL7 in colonies grown on 0.5% Impranil® plates.




PET_beads_acitivty_WT_vs_CrePHL7.csv: Comparison of enzymatic activity between wild-type (WT) Chlamydomonas reinhardtii and Cre PHL7 in degrading PET plastic beads.



Excel Files:





Plate_reader_settings.xlsx: Contains settings for the plate reader used to measure enzyme activity, absorbance, and fluorescence in the FDA and Impranil® assays.




Std_curve_TPA_100mM_and_500mM_phosphate_buffer.xlsx: Standard curve data for TPA quantification in phosphate buffer, essential for determining enzymatic degradation rates.




Transformation_cc1690_pJP32PHL7_Coloni&halo_count.xlsx: Data from transformation experiments involving the cc1690 strain expressing pJP32PHL7, including colony counts and halo formation.



Protein Sequencing Folder:

This folder contains detailed protein sequencing data from experiments analyzing the activity of the PHL7 enzyme, particularly in clearing zones of the plates. The subfolders PHL7_Bottom_clearing_zone and PHL7_Top_clearing_zone include visualizations, peptide reports, and data on protein identification from the enzyme secretion studies.

Raw Image Data:





Raw_Img/05_impr and Raw_Img/075_impr: Images documenting colony growth on 0.5% and 0.75% Impranil® plates. The images show clearing zones, indicative of enzymatic degradation of plastics by PHL7.



Time-Lapse Experiment:





Timelapse_Cre_PH7: This file contains a series of photos captured every 5 minutes over the course of 11 days, documenting a time-lapse experiment on TAP media agar plate with 0.5% Impranil®. The cells were arranged in a 96-well plate array. Throughout the experiment, the images show the gradual formation of clearing halos, especially in the later days, indicating secretion of active PHL7 enzyme by Chlamydomonas reinhardtii.