Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis
Abstract (−)-Carvone is a monoterpenoid with a spearmint flavor. A sustainable biotechnological production process for (−)-carvone is desirable. Although all enzymes in (−)-carvone biosynthesis have been functionally expressed in Escherichia coli independently, the yield was low in previous studies....
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Nature Portfolio
2021
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oai:doaj.org-article:5e010c658d71455cb769685da713f6452021-11-14T12:19:57ZIncreased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis10.1038/s41598-021-01469-y2045-2322https://doaj.org/article/5e010c658d71455cb769685da713f6452021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-01469-yhttps://doaj.org/toc/2045-2322Abstract (−)-Carvone is a monoterpenoid with a spearmint flavor. A sustainable biotechnological production process for (−)-carvone is desirable. Although all enzymes in (−)-carvone biosynthesis have been functionally expressed in Escherichia coli independently, the yield was low in previous studies. When cytochrome P450 limonene-6-hydroxylase (P450)/cytochrome P450 reductase (CPR) and carveol dehydrogenase (CDH) were expressed in a single strain, by-product formation (dihydrocarveol and dihydrocarvone) was detected. We hypothesized that P450 and CDH expression levels differ in E. coli. Thus, two strains independently expressing P450/CPR and CDH were mixed with different ratios, confirming increased carvone production and decreased by-product formation when CDH input was reduced. The optimum ratio of enzyme expression to maximize (−)-carvone production was determined using the proteome analysis quantification concatamer (QconCAT) method. Thereafter, a single strain expressing both P450/CPR and CDH was constructed to imitate the optimum expression ratio. The upgraded strain showed a 15-fold improvement compared to the initial strain, showing a 44 ± 6.3 mg/L (−)-carvone production from 100 mg/L (−)-limonene. Our study showed the usefulness of the QconCAT proteome analysis method for strain development in the industrial biotechnology field.Erika YoshidaMotoki KojimaMunenori SuzukiFumio MatsudaKazutaka ShimboAkiko OnukiYousuke NishioYoshihiro UsudaAkihiko KondoJun IshiiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021) |
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Medicine R Science Q Erika Yoshida Motoki Kojima Munenori Suzuki Fumio Matsuda Kazutaka Shimbo Akiko Onuki Yousuke Nishio Yoshihiro Usuda Akihiko Kondo Jun Ishii Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| description |
Abstract (−)-Carvone is a monoterpenoid with a spearmint flavor. A sustainable biotechnological production process for (−)-carvone is desirable. Although all enzymes in (−)-carvone biosynthesis have been functionally expressed in Escherichia coli independently, the yield was low in previous studies. When cytochrome P450 limonene-6-hydroxylase (P450)/cytochrome P450 reductase (CPR) and carveol dehydrogenase (CDH) were expressed in a single strain, by-product formation (dihydrocarveol and dihydrocarvone) was detected. We hypothesized that P450 and CDH expression levels differ in E. coli. Thus, two strains independently expressing P450/CPR and CDH were mixed with different ratios, confirming increased carvone production and decreased by-product formation when CDH input was reduced. The optimum ratio of enzyme expression to maximize (−)-carvone production was determined using the proteome analysis quantification concatamer (QconCAT) method. Thereafter, a single strain expressing both P450/CPR and CDH was constructed to imitate the optimum expression ratio. The upgraded strain showed a 15-fold improvement compared to the initial strain, showing a 44 ± 6.3 mg/L (−)-carvone production from 100 mg/L (−)-limonene. Our study showed the usefulness of the QconCAT proteome analysis method for strain development in the industrial biotechnology field. |
| format |
article |
| author |
Erika Yoshida Motoki Kojima Munenori Suzuki Fumio Matsuda Kazutaka Shimbo Akiko Onuki Yousuke Nishio Yoshihiro Usuda Akihiko Kondo Jun Ishii |
| author_facet |
Erika Yoshida Motoki Kojima Munenori Suzuki Fumio Matsuda Kazutaka Shimbo Akiko Onuki Yousuke Nishio Yoshihiro Usuda Akihiko Kondo Jun Ishii |
| author_sort |
Erika Yoshida |
| title |
Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| title_short |
Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| title_full |
Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| title_fullStr |
Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| title_full_unstemmed |
Increased carvone production in Escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| title_sort |
increased carvone production in escherichia coli by balancing limonene conversion enzyme expression via targeted quantification concatamer proteome analysis |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/5e010c658d71455cb769685da713f645 |
| work_keys_str_mv |
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1718429290137124864 |