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Expression of a hemA Gene from Agrobacterium radiobacter in a Rare Codon Optimizing Escherichia coli for Improving 5-aminolevulinate Production

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Abstract

The 5-aminolevulinate (ALA) synthase gene (hemA) from Agrobacterium radiobacter zju-0121, which was cloned previously in our laboratory, contains several rare codons. To enhance the expression of this gene, Escherichia coli Rosetta(DE3), which is a rare codon optimizer strain, was picked out as the host to construct an efficient recombinant strain. Cell extracts of the recombinant E. coli were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under the appropriate conditions. The results indicated that the activity of ALA synthase expressed in Rosetta(DE3)/pET-28a(+)-hemA was about 20% higher than that in E. coli BL21(DE3). Then the effects of precursors (glycine and succinate) and glucose, which is an inhibitor for ALA dehydratase as well as the carbon sources for cell growth, on the production of 5-aminolevulinate were investigated. Based on an optimal fed-batch culture system described in our previous work, up to 6.5 g/l (50 mM) ALA was produced in a 15-l fermenter.

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References

  1. Kennedy, J. C., Pottier, R. H., & Pross, D. C. (1990). Photodynamic therapy with endogenous protoporphyrin IX: basic principles and present clinical experience. Journal of Photochemistry and Photobiology B: Biology, 6, 143–148. doi:10.1016/1011-1344(90)85083-9.

    Article  CAS  Google Scholar 

  2. De Dominicis, C., Liberti, M., Perugia, G., De Nunzio, C., Sciobica, F., Zuccala, A., et al. (2001). Role of 5-aminolevulinic acid in the diagnosis and treatment of superficial bladder cancer: improvement in diagnosis sensitivity. Urology, 57, 1059–1062. doi:10.1016/S0090-4295(01)00948-7.

    Article  Google Scholar 

  3. Chen, H. M., Chen, C. H., Yang, H., Kuo, M. Y., & Kuo, Y. S. (2004). Successful treatment of oral verrucous hyperplasia with topical 5-aminolevulinic acid-mediated photodynamic therapy. Oral Oncology, 40, 630–637. doi:10.1016/j.oraloncology.2003.12.010.

    Article  CAS  Google Scholar 

  4. Sasikala, C. H., Ramana, V., & Raghuveer, R. P. (1994). 5-Aminolevulinic acid: a potential herbicide/insecticide from micro-organism. Biotechnology Progress, 10, 451–459. doi:10.1021/bp00029a001.

    Article  CAS  Google Scholar 

  5. Sasaki, K., Watanabe, M., Tanaka, T., & Tanaka, T. (2002). Biosynthesis, biotechnological production and applications of 5-aminolevulinic acid. Applied Microbiology and Biotechnology, 58, 23–29. doi:10.1007/s00253-001-0858-7.

    Article  CAS  Google Scholar 

  6. Vladimir, Y. B., Demain, A. L., & Zaitseva, N. I. (1997). The crucial contribution of starved resting cells to the elucidation of the pathway of vitamin B12 biosynthesis. Critical Reviews in Biotechnology, 17, 21–37. doi:10.3109/07388559709146605.

    Article  Google Scholar 

  7. Xie, L., Hall, D., Eiteman, M. A., & Altman, E. (2003). Optimization of recombinant aminolevulinate synthase production in Escherichia coli using factorial design. Applied Microbiology and Biotechnology, 63, 267–273. doi:10.1007/s00253-003-1388-2.

    Article  CAS  Google Scholar 

  8. Lee, D. H., Jun, W. J., Shin, D. H., Cho, H. Y., & Hong, B. S. (2005). Effect of culture conditions on production of 5-aminolevulinic acid by recombinant Escherichia coli. Bioscience, Biotechnology, and Biochemistry, 69, 470–476. doi:10.1271/bbb.69.470.

    Article  CAS  Google Scholar 

  9. Liu, X. X., Lin, J. P., Qin, G., & Cen, P. L. (2005). Expression a new hemA gene from Agrobacterium radiobacter in Escherichia coli for 5-aminolevulinate production. Chinese Journal of Chemical Engineering, 13, 522–528.

    CAS  Google Scholar 

  10. Chung, S. Y., Seo, K. H., & Rhee, J. I. (2005). Influence of culture conditions on the production of extra-cellular 5-aminolevulinic acid (ALA) by recombinant E. coli. Process Biochemistry, 40, 385–394. doi:10.1016/j.procbio.2004.01.024.

    Article  CAS  Google Scholar 

  11. Liu, X. X., Lin, J. P., & Cen, P. L. (2006). Effect of inducers on the production of 5-aminolevulinic acid by recombinant Escherichia coli. Preparative Biochemistry & Biotechnology, 36, 223–233. doi:10.1080/10826060600716638.

    Article  CAS  Google Scholar 

  12. Fu, W. Q., Lin, J. P., & Cen, P. L. (2007). 5-Aminolevulinate production with recombinant Escherichia coli using a rare codon optimizer host strain. Applied Microbiology and Biotechnology, 75, 777–782. doi:10.1007/s00253-007-0887-y.

    Article  CAS  Google Scholar 

  13. Drolet, M., & Sassarman, A. (1991). Cloning and nucleotide sequence of the hemA gene of Agrobacterium radiobacter. Molecular Genetics and Genomics, 226, 250–256.

    CAS  Google Scholar 

  14. Qin, G., Lin, J. P., Liu, X. X., & Cen, P. L. (2006). Effects of medium composition on production of 5-aminolevulinic acid by recombinant Escherichia coli. Journal of Bioscience and Bioengineering, 102, 316–322. doi:10.1263/jbb.102.316.

    Article  CAS  Google Scholar 

  15. Brinkmann, U., Mattes, R. E., & Buckel, P. (1989). High-level expression of recombinant genes in Escherichia coli is dependent on the availability of the dnaY gene product. Gene, 85, 109–114. doi:10.1016/0378-1119(89)90470-8.

    Article  CAS  Google Scholar 

  16. Seidel, H. M., Pompliano, D. L., & Knowles, J. R. (1992). Phosphonate biosynthesis: molecular cloning of the gene for phosphoenolpyruvate mutase from Tetrahymena pyriformis and overexpression of the gene product in Escherichia coli. Biochemistry, 31, 2598–2608. doi:10.1021/bi00124a021.

    Article  CAS  Google Scholar 

  17. Rosenberg, A. H., Goldman, E., Dunn, J. J., Studier, F. W., & Zubay, G. (1993). Effects of consecutive AGG codons on translation in Escherichia coli, demonstrated with a versatile codon test system. Journal of Bacteriology, 175, 716–722.

    CAS  Google Scholar 

  18. Nakamura, Y., Gojobori, T., & Ikemura, T. (2000). Codon usage tabulated from international DNA sequence databases: status for the year 2000. Nucleic Acids Research, 28, 292. doi:10.1093/nar/28.1.292.

    Article  CAS  Google Scholar 

  19. Lascelles, J. (1978). Regulation of pyrrole synthesis. In R. K. Clayton, & W. R. Sistrom (Eds.), Photosynthetic Bacteria (p. 795). New York: Plenum.

    Google Scholar 

  20. Lee, D. H., Jun, W. J., Kim, K. M., Shin, D. H., Cho, H. Y., & Hong, B. S. (2003). Inhibition of 5-aminolevulinic acid dehydratase in recombinant Escherichia coli using D-glucose. Enzyme and Microbial Technology, 32, 27–34. doi:10.1016/S0141-0229(02)00241-7.

    Article  CAS  Google Scholar 

  21. Fu, W. Q., Lin, J. P., & Cen, P. L. (2008). Enhancement of 5-aminolevulinate production with recombinant Escherichia coli using batch and fed-batch culture system. Bioresource Technology, 99, 4864–4870. doi:10.1016/j.biortech.2007.09.039.

    Article  CAS  Google Scholar 

  22. Burnham, B. F. (1970). σ-Aminolevulinic acid synthase (from Rhodopseudomonas sphaeroides). Methods in Enzymology, 17A, 195–204. doi:10.1016/0076-6879(71)17179-0.

    Article  Google Scholar 

  23. Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685. doi:10.1038/227680a0.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by a grant from the National Natural Science Foundation of China (20306026) and by a grant from the Ministry of Science and Technology of China (National Basic Research Program of China, 2007CB707805).

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  1. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, Zhejiang, 310027, China

    Weiqi Fu, Jianping Lin & Peilin Cen

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  1. Weiqi Fu
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  2. Jianping Lin
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Correspondence to Jianping Lin.

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Fu, W., Lin, J. & Cen, P. Expression of a hemA Gene from Agrobacterium radiobacter in a Rare Codon Optimizing Escherichia coli for Improving 5-aminolevulinate Production. Appl Biochem Biotechnol 160, 456–466 (2010). https://doi.org/10.1007/s12010-008-8363-4

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  • DOI: https://doi.org/10.1007/s12010-008-8363-4

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