Engineering the green algae Chlamydomonas incerta for recombinant protein production.
Chlamydomonas incerta, a genetically close relative of the model green alga Chlamydomonas reinhardtii, shows significant potential as a host for recombinant protein expression. Because of the close genetic relationship between C. incerta and C. reinhardtii, this species offers an additional referenc...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2025-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0321071 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850176171784798208 |
|---|---|
| author | Kalisa Kang Évellin do Espirito Santo Crisandra Jade Diaz Aaron Oliver Lisa Saxton Lauren May Stephen Mayfield João Vitor Dutra Molino |
| author_facet | Kalisa Kang Évellin do Espirito Santo Crisandra Jade Diaz Aaron Oliver Lisa Saxton Lauren May Stephen Mayfield João Vitor Dutra Molino |
| author_sort | Kalisa Kang |
| collection | DOAJ |
| description | Chlamydomonas incerta, a genetically close relative of the model green alga Chlamydomonas reinhardtii, shows significant potential as a host for recombinant protein expression. Because of the close genetic relationship between C. incerta and C. reinhardtii, this species offers an additional reference point for advancing our understanding of photosynthetic organisms, and also provides a potential new candidate for biotechnological applications. This study investigates C. incerta's capacity to express three recombinant proteins: the fluorescent protein mCherry, the hemicellulose-degrading enzyme xylanase, and the plastic-degrading enzyme PHL7. We have also examined the capacity to target protein expression to various cellular compartments in this alga, including the cytosol, secretory pathway, cytoplasmic membrane, and cell wall. When compared directly with C. reinhardtii, C. incerta exhibited a distinct but notable capacity for recombinant protein production. Cellular transformation with a vector encoding mCherry revealed that C. incerta produced approximately 3.5 times higher fluorescence levels and a 3.7-fold increase in immunoblot intensity compared to C. reinhardtii. For xylanase expression and secretion, both C. incerta and C. reinhardtii showed similar secretion capacities and enzymatic activities, with comparable xylan degradation rates, highlighting the industrial applicability of xylanase expression in microalgae. Finally, C. incerta showed comparable PHL7 activity levels to C. reinhardtii, as demonstrated by the in vitro degradation of a polyester polyurethane suspension, Impranil® DLN. Finally, we also explored the potential of cellular fusion for the generation of genetic hybrids between C. incerta and C. reinhardtii as a means to enhance phenotypic diversity and augment genetic variation. We were able to generate genetic fusion that could exchange both the recombinant protein genes, as well as associated selectable marker genes into recombinant offspring. These findings emphasize C. incerta's potential as a robust platform for recombinant protein production, and as a powerful tool for gaining a better understanding of microalgal biology. |
| format | Article |
| id | doaj-art-df9289c770874e5182be39c511c19245 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-df9289c770874e5182be39c511c192452025-08-20T02:19:18ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01204e032107110.1371/journal.pone.0321071Engineering the green algae Chlamydomonas incerta for recombinant protein production.Kalisa KangÉvellin do Espirito SantoCrisandra Jade DiazAaron OliverLisa SaxtonLauren MayStephen MayfieldJoão Vitor Dutra MolinoChlamydomonas incerta, a genetically close relative of the model green alga Chlamydomonas reinhardtii, shows significant potential as a host for recombinant protein expression. Because of the close genetic relationship between C. incerta and C. reinhardtii, this species offers an additional reference point for advancing our understanding of photosynthetic organisms, and also provides a potential new candidate for biotechnological applications. This study investigates C. incerta's capacity to express three recombinant proteins: the fluorescent protein mCherry, the hemicellulose-degrading enzyme xylanase, and the plastic-degrading enzyme PHL7. We have also examined the capacity to target protein expression to various cellular compartments in this alga, including the cytosol, secretory pathway, cytoplasmic membrane, and cell wall. When compared directly with C. reinhardtii, C. incerta exhibited a distinct but notable capacity for recombinant protein production. Cellular transformation with a vector encoding mCherry revealed that C. incerta produced approximately 3.5 times higher fluorescence levels and a 3.7-fold increase in immunoblot intensity compared to C. reinhardtii. For xylanase expression and secretion, both C. incerta and C. reinhardtii showed similar secretion capacities and enzymatic activities, with comparable xylan degradation rates, highlighting the industrial applicability of xylanase expression in microalgae. Finally, C. incerta showed comparable PHL7 activity levels to C. reinhardtii, as demonstrated by the in vitro degradation of a polyester polyurethane suspension, Impranil® DLN. Finally, we also explored the potential of cellular fusion for the generation of genetic hybrids between C. incerta and C. reinhardtii as a means to enhance phenotypic diversity and augment genetic variation. We were able to generate genetic fusion that could exchange both the recombinant protein genes, as well as associated selectable marker genes into recombinant offspring. These findings emphasize C. incerta's potential as a robust platform for recombinant protein production, and as a powerful tool for gaining a better understanding of microalgal biology.https://doi.org/10.1371/journal.pone.0321071 |
| spellingShingle | Kalisa Kang Évellin do Espirito Santo Crisandra Jade Diaz Aaron Oliver Lisa Saxton Lauren May Stephen Mayfield João Vitor Dutra Molino Engineering the green algae Chlamydomonas incerta for recombinant protein production. PLoS ONE |
| title | Engineering the green algae Chlamydomonas incerta for recombinant protein production. |
| title_full | Engineering the green algae Chlamydomonas incerta for recombinant protein production. |
| title_fullStr | Engineering the green algae Chlamydomonas incerta for recombinant protein production. |
| title_full_unstemmed | Engineering the green algae Chlamydomonas incerta for recombinant protein production. |
| title_short | Engineering the green algae Chlamydomonas incerta for recombinant protein production. |
| title_sort | engineering the green algae chlamydomonas incerta for recombinant protein production |
| url | https://doi.org/10.1371/journal.pone.0321071 |
| work_keys_str_mv | AT kalisakang engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT evellindoespiritosanto engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT crisandrajadediaz engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT aaronoliver engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT lisasaxton engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT laurenmay engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT stephenmayfield engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction AT joaovitordutramolino engineeringthegreenalgaechlamydomonasincertaforrecombinantproteinproduction |