Abstract
Objectives
Putative genes crtE, crtB, and crtI from Deinococcus wulumiqiensis R12, a novel species, were identified by genome mining and were co-expressed using the optimized Shine-Dalgarno (SD) regions to improve lycopene yield.
Results
A lycopene biosynthesis pathway was constructed by co-expressing these three genes in Escherichia coli. After optimizing the upstream SD regions and the culture medium, the recombinant strain EDW11 produced 88 mg lycopene g−1 dry cell wt (780 mg lycopene l−1) after 40 h fermentation without IPTG induction, while the strain EDW without optimized SD regions only produced 49 mg lycopene g−1 dry cell wt (417 mg lycopene l−1).
Conclusion
Based on the optimization of the upstream SD regions and culture medium, the yield of the strain EDW11 reached a high level during microbial lycopene production until now.
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Acknowledgments
This work was supported by the National Science Foundation for Distinguished Young Scholars of China (21225626), the National High Technology Research and Development Program of China (2012AA021705, 2012AA022101), the National Natural Science Foundation of China for Young Scholars (21406111), and the Natural Science Foundation of Jiangsu Province, China (BK20131406).
Supporting information
Supplementary Table 1—Bacterial strains and plasmids used in this work.
Supplementary Table 2—Primers used in this work.
Supplementary Fig. 1—HPLC profiles of the isolated carotenoids from the strain EDWe, EDW, EDW11, EDW12, EDW21, EDW22, and the commercial lycopene.
Supplementary Fig. 2—The color of the cell pellets of the strains EDW11, EDW12, EDW21, and EDW22.
Supplementary Fig. 3—Comparison of the phytoene dehydrogenase expression and the growth of the recombinant strains E. coli EDW-I1 and EDW-I2.
Supplementary Fig. 4—Comparison of the phytoene synthase expression and the growth of the recombinant strains E. coli EDW-B1 and EDW-B2.
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Weiyue Jin and Xian Xu have contributed to the work equally and should be regarded as co-first authors.
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Jin, W., Xu, X., Jiang, L. et al. Putative carotenoid genes expressed under the regulation of Shine–Dalgarno regions in Escherichia coli for efficient lycopene production. Biotechnol Lett 37, 2303–2310 (2015). https://doi.org/10.1007/s10529-015-1922-1
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DOI: https://doi.org/10.1007/s10529-015-1922-1