Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin
Abstract Background Crocetin is a multifunctional apocarotenoid natural product derived from saffron, holding significant promises for protection against various diseases and other nutritional applications. Historically, crocetin has been extracted from saffron stigmas, but this method is hindered b...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2025-01-01
|
| Series: | Biotechnology for Biofuels and Bioproducts |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-024-02598-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841544899669786624 |
|---|---|
| author | Tingan Zhou Young-Kyoung Park Jing Fu Piotr Hapeta Cinzia Klemm Rodrigo Ledesma-Amaro |
| author_facet | Tingan Zhou Young-Kyoung Park Jing Fu Piotr Hapeta Cinzia Klemm Rodrigo Ledesma-Amaro |
| author_sort | Tingan Zhou |
| collection | DOAJ |
| description | Abstract Background Crocetin is a multifunctional apocarotenoid natural product derived from saffron, holding significant promises for protection against various diseases and other nutritional applications. Historically, crocetin has been extracted from saffron stigmas, but this method is hindered by the limited availability of high-quality raw materials and complex extraction processes. To overcome these challenges, metabolic engineering and synthetic biology can be applied to the sustainable production of crocetin. Results We constructed a Yarrowia lipolytica strain using hybrid promoters and copy number adjustment, which was able to produce 2.66 g/L of β-carotene, the precursor of crocetin. Next, the crocetin biosynthetic pathway was introduced, and we observed both the production and secretion of crocetin. Subsequently, the metabolite profiles under varied temperatures were studied and we found that low temperature was favorable for crocetin biosynthesis in Y. lipolytica. Therefore, a two-step temperature-shift fermentation strategy was adopted to optimize yeast growth and biosynthetic enzyme activity, bringing a 2.3-fold increase in crocetin titer. Lastly, fermentation media was fine-tuned for an optimal crocetin output of 30.17 mg/L, bringing a 51% higher titer compared with the previous highest report in shake flasks. Concomitantly, we also generated Y. lipolytica strains capable of achieving substantial zeaxanthin production, yielding 1575.09 mg/L, doubling the previous highest reported titer. Conclusions Through metabolic engineering and fermentation optimization, we demonstrated the first de novo biosynthesis of crocetin in the industrial yeast Yarrowia lipolytica. In addition, we achieved a higher crocetin titer in flasks than all our known reports. This work not only represents a high production of crocetin, but also entails a significant simultaneous zeaxanthin production, setting the stage for sustainable and cost-effective production of these valuable compounds. |
| format | Article |
| id | doaj-art-a2f9cca7574f49be84e1f3fcfbf77fe3 |
| institution | Kabale University |
| issn | 2731-3654 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj-art-a2f9cca7574f49be84e1f3fcfbf77fe32025-01-12T12:11:40ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542025-01-0118111210.1186/s13068-024-02598-yMetabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetinTingan Zhou0Young-Kyoung Park1Jing Fu2Piotr Hapeta3Cinzia Klemm4Rodrigo Ledesma-Amaro5Department of Bioengineering, Imperial College Centre for Synthetic Biology, Bezos Centre for Sustainable Protein, UKRI Engineering Biology Mission Hub on Microbial Food, Imperial College LondonDepartment of Bioengineering, Imperial College Centre for Synthetic Biology, Bezos Centre for Sustainable Protein, UKRI Engineering Biology Mission Hub on Microbial Food, Imperial College LondonDepartment of Bioengineering, Imperial College Centre for Synthetic Biology, Bezos Centre for Sustainable Protein, UKRI Engineering Biology Mission Hub on Microbial Food, Imperial College LondonDepartment of Bioengineering, Imperial College Centre for Synthetic Biology, Bezos Centre for Sustainable Protein, UKRI Engineering Biology Mission Hub on Microbial Food, Imperial College LondonDepartment of Bioengineering, Imperial College Centre for Synthetic Biology, Bezos Centre for Sustainable Protein, UKRI Engineering Biology Mission Hub on Microbial Food, Imperial College LondonDepartment of Bioengineering, Imperial College Centre for Synthetic Biology, Bezos Centre for Sustainable Protein, UKRI Engineering Biology Mission Hub on Microbial Food, Imperial College LondonAbstract Background Crocetin is a multifunctional apocarotenoid natural product derived from saffron, holding significant promises for protection against various diseases and other nutritional applications. Historically, crocetin has been extracted from saffron stigmas, but this method is hindered by the limited availability of high-quality raw materials and complex extraction processes. To overcome these challenges, metabolic engineering and synthetic biology can be applied to the sustainable production of crocetin. Results We constructed a Yarrowia lipolytica strain using hybrid promoters and copy number adjustment, which was able to produce 2.66 g/L of β-carotene, the precursor of crocetin. Next, the crocetin biosynthetic pathway was introduced, and we observed both the production and secretion of crocetin. Subsequently, the metabolite profiles under varied temperatures were studied and we found that low temperature was favorable for crocetin biosynthesis in Y. lipolytica. Therefore, a two-step temperature-shift fermentation strategy was adopted to optimize yeast growth and biosynthetic enzyme activity, bringing a 2.3-fold increase in crocetin titer. Lastly, fermentation media was fine-tuned for an optimal crocetin output of 30.17 mg/L, bringing a 51% higher titer compared with the previous highest report in shake flasks. Concomitantly, we also generated Y. lipolytica strains capable of achieving substantial zeaxanthin production, yielding 1575.09 mg/L, doubling the previous highest reported titer. Conclusions Through metabolic engineering and fermentation optimization, we demonstrated the first de novo biosynthesis of crocetin in the industrial yeast Yarrowia lipolytica. In addition, we achieved a higher crocetin titer in flasks than all our known reports. This work not only represents a high production of crocetin, but also entails a significant simultaneous zeaxanthin production, setting the stage for sustainable and cost-effective production of these valuable compounds.https://doi.org/10.1186/s13068-024-02598-yCrocetinZeaxanthinβ-CaroteneYarrowia lipolyticaSecretory biosynthesisMetabolic engineering |
| spellingShingle | Tingan Zhou Young-Kyoung Park Jing Fu Piotr Hapeta Cinzia Klemm Rodrigo Ledesma-Amaro Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin Biotechnology for Biofuels and Bioproducts Crocetin Zeaxanthin β-Carotene Yarrowia lipolytica Secretory biosynthesis Metabolic engineering |
| title | Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin |
| title_full | Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin |
| title_fullStr | Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin |
| title_full_unstemmed | Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin |
| title_short | Metabolic engineering of Yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin |
| title_sort | metabolic engineering of yarrowia lipolytica for the production and secretion of the saffron ingredient crocetin |
| topic | Crocetin Zeaxanthin β-Carotene Yarrowia lipolytica Secretory biosynthesis Metabolic engineering |
| url | https://doi.org/10.1186/s13068-024-02598-y |
| work_keys_str_mv | AT tinganzhou metabolicengineeringofyarrowialipolyticafortheproductionandsecretionofthesaffroningredientcrocetin AT youngkyoungpark metabolicengineeringofyarrowialipolyticafortheproductionandsecretionofthesaffroningredientcrocetin AT jingfu metabolicengineeringofyarrowialipolyticafortheproductionandsecretionofthesaffroningredientcrocetin AT piotrhapeta metabolicengineeringofyarrowialipolyticafortheproductionandsecretionofthesaffroningredientcrocetin AT cinziaklemm metabolicengineeringofyarrowialipolyticafortheproductionandsecretionofthesaffroningredientcrocetin AT rodrigoledesmaamaro metabolicengineeringofyarrowialipolyticafortheproductionandsecretionofthesaffroningredientcrocetin |