Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

doi:10.1007/s13659-018-0172-z

Natural Products and Bioprospecting ›› 2018, Vol. 8 ›› Issue (5): 397-403.DOI: 10.1007/s13659-018-0172-z

• ORIGINAL ARTICLES • Previous Articles

Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

Shun Li^1,2, Jun-Chao Huang¹

1 Department of Economic Plants and Biotechnology, Yunnan Key Laboratory for Wild Plant Resources, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, People's Republic of China;
2 University of Chinese Academy of Sciences, Beijing 100049, People's Republic of China

Received:2018-04-25 Revised:2018-05-30 Online:2018-09-19 Published:2018-10-24
Contact: Jun-Chao Huang
Supported by:
This study was supported by a research grant from Department of Economic Plants and Biotechnology, Yunnan Key Laboratory for Wild Plant Resources, Kunming Institute of Botany, Chinese Academy of Sciences.

Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

Shun Li^1,2, Jun-Chao Huang¹

1 Department of Economic Plants and Biotechnology, Yunnan Key Laboratory for Wild Plant Resources, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming 650201, People's Republic of China;
2 University of Chinese Academy of Sciences, Beijing 100049, People's Republic of China

通讯作者: Jun-Chao Huang
基金资助:
This study was supported by a research grant from Department of Economic Plants and Biotechnology, Yunnan Key Laboratory for Wild Plant Resources, Kunming Institute of Botany, Chinese Academy of Sciences.

Abstract

Abstract: Astaxanthin is a value-added ketocarotenoid with great potential in nutraceutical and pharmaceutical industries. Genetic engineering of heterologous hosts for astaxanthin production has attracted great attention. In this study, we assessed some key factors, including codon usage of the expressed genes, types of promoters, bacterial strains, and culture media, for engineered Escherichia coli to produce astaxanthin. The effect of codon usage was shown to be related to the types of promoters. E. coli DH5α was superior to other strains for astaxanthin production. Different culture media greatly affected the contents and yields of astaxanthin in engineered E. coli. When the expression cassette containing GadE promoter and its driving genes, HpCHY and CrBKT, was inserted into the plasmid pACCAR16△crtX and expressed in E. coli DH5α, the engineered strain was able to produce 4.30 ±0.28 mg/g dry cell weight (DCW) or 24.16 ±2.03 mg/L of astaxanthin, which was a sevenfold or 40-fold increase over the initial production of 0.62 ±0.03 mg/g DCW or 0.61 ±0.05 mg/L.

Key words: Astaxanthin, Escherichia coli, Bacterial strains, Codon usage, Medium

摘要： Astaxanthin is a value-added ketocarotenoid with great potential in nutraceutical and pharmaceutical industries. Genetic engineering of heterologous hosts for astaxanthin production has attracted great attention. In this study, we assessed some key factors, including codon usage of the expressed genes, types of promoters, bacterial strains, and culture media, for engineered Escherichia coli to produce astaxanthin. The effect of codon usage was shown to be related to the types of promoters. E. coli DH5α was superior to other strains for astaxanthin production. Different culture media greatly affected the contents and yields of astaxanthin in engineered E. coli. When the expression cassette containing GadE promoter and its driving genes, HpCHY and CrBKT, was inserted into the plasmid pACCAR16△crtX and expressed in E. coli DH5α, the engineered strain was able to produce 4.30 ±0.28 mg/g dry cell weight (DCW) or 24.16 ±2.03 mg/L of astaxanthin, which was a sevenfold or 40-fold increase over the initial production of 0.62 ±0.03 mg/g DCW or 0.61 ±0.05 mg/L.

关键词: Astaxanthin, Escherichia coli, Bacterial strains, Codon usage, Medium

Shun Li, Jun-Chao Huang. Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin[J]. Natural Products and Bioprospecting, 2018, 8(5): 397-403.

Shun Li, Jun-Chao Huang. Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin[J]. 应用天然产物, 2018, 8(5): 397-403.

References

1. R.R. Ambati, S.M. Phang, S. Ravi, R.G. Aswathanarayana, Mar. Drugs 12, 128-152 (2014)
2. J.P. Yuan, J. Peng, K. Yin, J.H. Wang, Mol. Nutr. Food Res. 55, 150-165 (2011)
3. I. Higuera-Ciapara, L. Félix-Valenzuela, F.M. Goycoolea, Crit. Rev. Food Sci. Nutr. 46, 185-196 (2006)
4. J. Li, D.L. Zhu, J.F. Niu, S.D. Shen, G.C. Wang, Biotechnol. Adv. 29, 568-574 (2011)
5. C.W. Wang, M.K. Oh, J.C. Liao, Biotechnol. Bioeng. 62, 235-241 (1999)
6. S.K. Choi, S. Matsuda, T. Hoshino, X. Peng, N. Misawa, Appl. Microbiol. Biotechnol. 72, 1238-1246 (2006)
7. M.A. Scaife, A.M. Burja, P.C. Wright, Biotechnol. Bioeng. 103, 944-955 (2009)
8. K. Lemuth, K. Steuer, C. Albermann, Microb. Cell Fact. 10, 1-12 (2011)
9. L. Zelcbuch, N. Antonovsky, A. Bar-Even, A. Levin-Karp, U. Barenholz, M. Dayagi, W. Liebermeister, A. Flamholz, E. Noor, S. Amram, A. Brandis, T. Bareia, I. Yofe, H. Jubran, R. Milo, Nucleic Acids Res. 41, e98-e98 (2013)
10. T. Ma, Y.J. Zhou, X.W. Li, F.Y. Zhu, Y.B. Cheng, Y. Liu, Z.X. Deng, T.G. Liu, Biotechnol. J. 11, 228-237 (2016)
11. Q. Lu, Y.F. Bu, J.Z. Liu, Mar. Drugs 15, 296 (2017)
12. P.P. Zhou, L.D. Ye, W.P. Xie, X.M. Lv, H.W. Yu, Appl. Microbiol. Biotechnol. 99, 8419-8428 (2015)
13. P.P. Zhou, W.P. Xie, A.P. Li, F. Wang, Z. Yao, Q. Bian, Y.Q. Zhu, H.W. Yu, L.D. Ye, Enzyme Microb. Technol. 100, 28-36 (2017)
14. N. Misawa, Y. Satomi, K. Kondo, A. Yokoyama, S. Kajiwara, T. Saito, T. Ohtani, W. Miki, J. Bacteriol. 177, 6575-6584 (1995)
15. J.C. Huang, Y.J. Zhong, J. Liu, G. Sandmann, F. Chen, Metab. Eng. 17, 59-67 (2013)
16. G. Boël, R. Letso, H. Neely, W.N. Price, K.H. Wong, M. Su, J.D. Luff, M. Valecha, J.K. Everett, T.B. Acton, R. Xiao, G.T. Montelione, D.P. Aalberts, J.F. Hunt, Nature 529, 358-363 (2016)
17. F. Hommais, E. Krin, J.Y. Coppe é, C. Lacroix, E. Yeramian, A. Danchin, P. Bertin, Microbiology 150, 61-72 (2004)
18. Z. Ma, S.M. Gong, H. Richard, D.L. Tucker, T. Conway, J.W. Foster, Mol. Microbiol. 49, 1309-1320 (2003)
19. R.H. Dahl, F.Z. Zhang, J. Alonso-Gutierrez, E. Baidoo, T.S. Batth, A.M. Redding-Johanson, C.J. Petzold, A. Mukhopadhyay, T.S. Lee, P.D. Adams, J.D. Keasling, Nat. Biotechnol. 31, 1039-1046 (2013)
20. A.K. Sayed, J.W. Foster, Mol. Microbiol. 71, 1435-1450 (2009)
21. H. Richard, J.W. Foster, Microbiology 153, 3154-3161 (2007)
22. A. Ruther, N. Misawa, P. Böger, G. Sandmann, Appl. Microbiol. Biotechnol. 48, 162-167 (1997)
23. H.S. Chae, K.H. Kim, S.C. Kim, P.C. Lee, Appl. Biochem. Biotechnol. 162, 2333-2344 (2010)
24. J.M. Monk, A. Koza, M.A. Campodonico, D. Machado, J.M. Seoane, B.O. Palsson, M.J. Herrgård, A.M. Feist, Cell Syst. 3, 238-251 (2016)
25. J.M. Woo, J.W. Kim, J.W. Song, L.M. Blank, J.B. Park, PLoS ONE 11, e0163265 (2016)
26. J. Shiloach, R. Fass, Biotechnol. Adv. 23, 345-357 (2005)
27. W.P. Huang, J.R. Ye, J.J. Zhang, Y. Lin, M.X. He, J.C. Huang, Algal Res. 17, 236-243 (2016)

Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 1

Recommended Articles

Metrics

Comments