Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i>
The sesquiterpene (+)-zizaene is the direct precursor of khusimol, the main fragrant compound of the vetiver essential oil from <i>Chrysopogon zizanioides</i> and used in nearly 20% of men’s fine perfumery. The biotechnological production of such fragrant sesquiterpenes is a pr...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-09-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/24/18/3356 |
| _version_ | 1818174662805291008 |
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| author | Francisco Aguilar Thomas Scheper Sascha Beutel |
| author_facet | Francisco Aguilar Thomas Scheper Sascha Beutel |
| author_sort | Francisco Aguilar |
| collection | DOAJ |
| description | The sesquiterpene (+)-zizaene is the direct precursor of khusimol, the main fragrant compound of the vetiver essential oil from <i>Chrysopogon zizanioides</i> and used in nearly 20% of men’s fine perfumery. The biotechnological production of such fragrant sesquiterpenes is a promising alternative towards sustainability; nevertheless, product recovery from fermentation is one of the main constraints. In an effort to improve the (+)-zizaene recovery from a metabolically-engineered <i>Escherichia coli</i>, we developed an integrated bioprocess by coupling fermentation and (+)-zizaene recovery using adsorber extractants. Initially, (+)-zizaene volatilization was confirmed from cultivations with no extractants but application of liquid−liquid phase partitioning cultivation (LLPPC) improved (+)-zizaene recovery nearly 4-fold. Furthermore, solid−liquid phase partitioning cultivation (SLPPC) was evaluated by screening polymeric adsorbers, where Diaion HP20 reached the highest recovery. Bioprocess was scaled up to 2 L bioreactors and in situ recovery configurations integrated to fermentation were evaluated. External recovery configuration was performed with an expanded bed adsorption column and improved (+)-zizaene titers 2.5-fold higher than LLPPC. Moreover, internal recovery configuration (IRC) further enhanced the (+)-zizaene titers 2.2-fold, whereas adsorption velocity was determined as critical parameter for recovery efficiency. Consequently, IRC improved the (+)-zizaene titer 8.4-fold and productivity 3-fold from our last report, achieving a (+)-zizaene titer of 211.13 mg L<sup>−1</sup> and productivity of 3.2 mg L<sup>−1</sup> h<sup>−1</sup>. This study provides further knowledge for integration of terpene bioprocesses by in situ product recovery, which could be applied for many terpene studies towards the industrialization of fragrant molecules. |
| first_indexed | 2024-12-11T19:47:58Z |
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| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-11T19:47:58Z |
| publishDate | 2019-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-01632fdc1b2c4237b6f8061ec5b8cba22022-12-22T00:52:50ZengMDPI AGMolecules1420-30492019-09-012418335610.3390/molecules24183356molecules24183356Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i>Francisco Aguilar0Thomas Scheper1Sascha Beutel2Institute of Technical Chemistry, Leibniz University of Hannover, Callinstr. 5, 30167 Hannover, GermanyInstitute of Technical Chemistry, Leibniz University of Hannover, Callinstr. 5, 30167 Hannover, GermanyInstitute of Technical Chemistry, Leibniz University of Hannover, Callinstr. 5, 30167 Hannover, GermanyThe sesquiterpene (+)-zizaene is the direct precursor of khusimol, the main fragrant compound of the vetiver essential oil from <i>Chrysopogon zizanioides</i> and used in nearly 20% of men’s fine perfumery. The biotechnological production of such fragrant sesquiterpenes is a promising alternative towards sustainability; nevertheless, product recovery from fermentation is one of the main constraints. In an effort to improve the (+)-zizaene recovery from a metabolically-engineered <i>Escherichia coli</i>, we developed an integrated bioprocess by coupling fermentation and (+)-zizaene recovery using adsorber extractants. Initially, (+)-zizaene volatilization was confirmed from cultivations with no extractants but application of liquid−liquid phase partitioning cultivation (LLPPC) improved (+)-zizaene recovery nearly 4-fold. Furthermore, solid−liquid phase partitioning cultivation (SLPPC) was evaluated by screening polymeric adsorbers, where Diaion HP20 reached the highest recovery. Bioprocess was scaled up to 2 L bioreactors and in situ recovery configurations integrated to fermentation were evaluated. External recovery configuration was performed with an expanded bed adsorption column and improved (+)-zizaene titers 2.5-fold higher than LLPPC. Moreover, internal recovery configuration (IRC) further enhanced the (+)-zizaene titers 2.2-fold, whereas adsorption velocity was determined as critical parameter for recovery efficiency. Consequently, IRC improved the (+)-zizaene titer 8.4-fold and productivity 3-fold from our last report, achieving a (+)-zizaene titer of 211.13 mg L<sup>−1</sup> and productivity of 3.2 mg L<sup>−1</sup> h<sup>−1</sup>. This study provides further knowledge for integration of terpene bioprocesses by in situ product recovery, which could be applied for many terpene studies towards the industrialization of fragrant molecules.https://www.mdpi.com/1420-3049/24/18/3356(+)-zizaenekhusimenekhusimolvetiver essential oilin situ product recoveryexpanded bed adsorptionsesquiterpenesterpenes<i>Chrysopogon zizanioides</i> |
| spellingShingle | Francisco Aguilar Thomas Scheper Sascha Beutel Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i> Molecules (+)-zizaene khusimene khusimol vetiver essential oil in situ product recovery expanded bed adsorption sesquiterpenes terpenes <i>Chrysopogon zizanioides</i> |
| title | Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i> |
| title_full | Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i> |
| title_fullStr | Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i> |
| title_full_unstemmed | Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i> |
| title_short | Improved Production and In Situ Recovery of Sesquiterpene (+)-Zizaene from Metabolically-Engineered <i>E. coli</i> |
| title_sort | improved production and in situ recovery of sesquiterpene zizaene from metabolically engineered i e coli i |
| topic | (+)-zizaene khusimene khusimol vetiver essential oil in situ product recovery expanded bed adsorption sesquiterpenes terpenes <i>Chrysopogon zizanioides</i> |
| url | https://www.mdpi.com/1420-3049/24/18/3356 |
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