Presentation Information
[3SBT-18]Fermentative Production of Olive-Derived Polyphenols Using a Chromosome-Engineered E. coli Platform Strain for Aromatic Compound Production
○Daisuke Koma1, Ryosuke Fujiwara1, Hiroyuki Ohashi1, Takashi Ohmoto1, Hayato Yamanaka1, Yasuharu Sato2, Takao Ohashi3, Ryo Misaki4 (1. Osaka Research Institute of Industrial Science and Technology (Japan), 2. Hokkaido Univ. (Japan), 3. Setsunan Univ. (Japan), 4. UOsaka (Japan))
Keywords:
aromatic compound,hydroxytyrosol,UGT,chromosome,olive
[Purpose]
3-hydroxytyrosol (HT) is a natural polyphenol found mainly in Olea europaea (olive). It is a potent antioxidant that protects human cells from reactive oxygen species (ROS), and various physiological activities have been reported. HT is currently used in cosmetics, dietary supplements, and pharmaceuticals. Fermentative production of HT is highly attractive due to its potential to reduce production costs; therefore, we aimed to generate strains capable of producing HT and its derivatives.
[Methods]
We have developed a Phe-producing strain that serves as a platform for the fermentative production of a variety of aromatic compounds. A key feature of this strain is that all pathway genes are integrated into the chromosome under the control of the T7 expression system. For Phe production, the titer exceeded 75 g/L in a 3-L jar fermenter. This platform strain was subsequently used as the host for constructing HT-producing strains.
[Results]
Using the platform strain, an HT-producing strain was constructed by integrating HT biosynthetic pathway genes into the chromosome along with T7 promoters. After several rounds of optimization, the HT titer increased to more than 10 g/L in a 3-L jar fermenter.
In addition, we focused on hydroxytyrosol 4-O-glucoside (HT4G), which is more chemically stable than HT. By introducing several genes encoding UDP-dependent glycosyltransferases (UGTs) on plasmids, the HT-producing strain was readily converted into strains capable of producing hydroxytyrosol 1-O-glucoside, hydroxytyrosol 3-O-glucoside, and HT4G. Among these, the best HT4G-producing strain achieved a titer of approximately 10 g/L in a 3-L jar fermenter.
[Conclusion]
We applied the chromosome-engineered E. coli platform strain to the fermentative production of olive-derived polyphenols. This platform enables high-level production of HT in a plasmid-free manner and can be readily extended to HT derivative production, as demonstrated by HT4G production via plasmid-based UGT expression. The improved stability of HT4G further highlights its potential for industrial applications. In addition, these results demonstrate the versatility and scalability of this platform system for biomanufacturing.
[Acknowledgments]
This study was supported by the JST Adaptable and Seamless Technology Transfer Program through Target-driven R&D (A-STEP), Grant Numbers JPMJTM19CC, JPMJTM20QJ and JPMJTR23U4, and also partially supported by JSPS KAKENHI Grant Numbers 19780082 and 21780105.
3-hydroxytyrosol (HT) is a natural polyphenol found mainly in Olea europaea (olive). It is a potent antioxidant that protects human cells from reactive oxygen species (ROS), and various physiological activities have been reported. HT is currently used in cosmetics, dietary supplements, and pharmaceuticals. Fermentative production of HT is highly attractive due to its potential to reduce production costs; therefore, we aimed to generate strains capable of producing HT and its derivatives.
[Methods]
We have developed a Phe-producing strain that serves as a platform for the fermentative production of a variety of aromatic compounds. A key feature of this strain is that all pathway genes are integrated into the chromosome under the control of the T7 expression system. For Phe production, the titer exceeded 75 g/L in a 3-L jar fermenter. This platform strain was subsequently used as the host for constructing HT-producing strains.
[Results]
Using the platform strain, an HT-producing strain was constructed by integrating HT biosynthetic pathway genes into the chromosome along with T7 promoters. After several rounds of optimization, the HT titer increased to more than 10 g/L in a 3-L jar fermenter.
In addition, we focused on hydroxytyrosol 4-O-glucoside (HT4G), which is more chemically stable than HT. By introducing several genes encoding UDP-dependent glycosyltransferases (UGTs) on plasmids, the HT-producing strain was readily converted into strains capable of producing hydroxytyrosol 1-O-glucoside, hydroxytyrosol 3-O-glucoside, and HT4G. Among these, the best HT4G-producing strain achieved a titer of approximately 10 g/L in a 3-L jar fermenter.
[Conclusion]
We applied the chromosome-engineered E. coli platform strain to the fermentative production of olive-derived polyphenols. This platform enables high-level production of HT in a plasmid-free manner and can be readily extended to HT derivative production, as demonstrated by HT4G production via plasmid-based UGT expression. The improved stability of HT4G further highlights its potential for industrial applications. In addition, these results demonstrate the versatility and scalability of this platform system for biomanufacturing.
[Acknowledgments]
This study was supported by the JST Adaptable and Seamless Technology Transfer Program through Target-driven R&D (A-STEP), Grant Numbers JPMJTM19CC, JPMJTM20QJ and JPMJTR23U4, and also partially supported by JSPS KAKENHI Grant Numbers 19780082 and 21780105.
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