Design of the general concept of regulated inactivation of the gene shikimatkinase for modification of the shikimat way in bacteria Bacillus subtilis

  • Yu Chao Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus
  • Maria Y. Shonina Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus
  • Aliaksei V. Lahodzich Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

Abstract

In this work demonstrated the possibility of the transformation strains of B. subtilis used to create strains-producers of shikimic acid. For 6 strains of B. subtilis, with the ability to increased synthesis of tryptophan, produced amplicons region of the genome containing genes tmrB and aroI. RFLP-analysis of the received amplicons showed their genetic polymorphism, which may be the result of mutagenesis used when receiving strains-producers of tryptophan. Based on the PCR method designed a system for fast screening of transformants for the presence of recombination and genome rearrangements presence integrative structure comprising bacterial strains used chromosomes from transformants.

Author Biographies

Yu Chao, Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

postgraduate student at the department of genetics, faculty of biology

Maria Y. Shonina, Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

postgraduate student at the department of genetics, faculty of biology

Aliaksei V. Lahodzich, Belarusian State University, Niezaliežnasci Avenue, 4, 220030, Minsk, Belarus

PhD (biology), docent; associate professor at the department of genetics, faculty of biology

References

  1. Zhang Y., Liu A., Ye Z. G., et al. New Approach to the Total Synthesis of (–)-Zeylenone from Shikimic Acid. Chem. & Pharm. Bull. 2006. Vol. 54, No. 10. P. 1459 –1461. 2. Bradley D. Star role for bacteria in controlling flu pandemic? Nature Rev.: Drug Discovery. 2005. Vol. 4, No. 12. P. 945– 946.
  2. Mair H.-J. Process for the preparation of shikimic acid its derivatives. US006130354 (A). 2000-10-10.
  3. Raghavendra T. R., Vaidyanathan P., Swathi H. K., et al. Prospecting for alternate sources of shikimic acid, a precursor of Tamiflu, a bird-flu drug. Current sci. 2009. Vol. 96, No. 6. P. 771–772.
  4. Brazdova B., Tan N. S., Samoshina N. M., et al. Novel easily accessible glucosidase inhibitors: 4-hydroxy-5-alkoxy-1,2-cyclohexanedicarboxylic acids. Carbohydrate Research. 2008. Vol. 344, No. 3. P. 311–321.
  5. Lingens F. The Biosynthesis of Aromatic Amino Acids and its Regulation. Angewandte Chem. Int. Ed. 2003. Vol. 7, No. 5. P. 350 –360.
  6. Ruohong S. Separation of Shikimic Acid from Pine Needles. Chem. Engineering & Technology. 2008. Vol. 31, No. 3. P. 469 – 473.
  7. Shinada T., Yoshida Y., Ohfune Y. Direct conversion of 1,2-diol into allyl sulfide. Regioselective transformation of (–)-quinic acid to (–)-shikimic acid. Tetrahedron Letters. 1998. Vol. 39, No. 33. P. 6027–6028.
  8. Adachi O., Ano Y., Toyama H., et al. High shikimate production from quinate with two enzymatic systems of acetic acid bacteria. Biosci., Biotech., a. Biochem. 2006. Vol. 70, No. 10. P. 2579 –2582.
  9. Taylor R. G., Walker D. C., McInnes R. R. E. coli host strains significantly affect the quality of small scale plasmid DNA preparations used for sequencing. Nucleic Acids Research. 1993. Vol. 21, No. 7. P. 1677–1678.
  10. Bullock W. O., Fernandez J. M., Short J. M. XL1-blue: a high efficiency plasmid transforming recA Escherichia coli strain with beta-galactosidase selection. BioTech. 1987. Vol. 5, No. 3. P. 376–379.
  11. Anagnostopoulos C., Spizizen J. Requirements for transformation in Bacillus subtilis. J. of Bacteriology. 1961. Vol. 81, No. 5. P. 741–746.
  12. Senatorova V. N. The study of the process and creation of technology of biosynthesis of L-tryptophan strain-producer Bacillus subtilis Of-15 : dissertatsiya ... kandidata tech. nauk : 03.00.23. Moscow, 2000 (in Russ.).
  13. Lahodzich A.V. Plasmids of pBS72 family as a basis for creation of vector systems. Biotechnology of the Future: EU-Russia: Prospects for Cooperation in Biotechnology in the Seventh Framework Programme : affiliated to the Int. symp. (Saint Petersburg, 5–8 June, 2006). Moscow, 2006. P. 47– 48.
  14. Sambrook J., Fritsch E., Maniatis T. Molecular cloning : a laboratory manual. 2nd ed. New York : Cold Spring Harbor Lab. : Cold Spring Harbor Publishing, 1989.
  15. Birnboim H. L., Doly J. A rapid alkaline extraction procedure for screening of recombinant plasmid DNA. Nucleic Acids Research. 1979. Vol. 7, No. 6. P. 1513–1523. 17. Schallmey M., Singh A., Ward O. P. Developments in the use of Bacillus species for industrial production. Canad. J. Microbiol. 2004. Vol. 50, No. 1. P. 1–17.
  16. Kunst F., Ogasawara N., Moszer I. The complete genome sequence of the gram-positive bacterium Bacillus subtilis. Nature. 1997. Vol. 390, No. 6657. P. 249 –256. 19. Bacillus subtilis 168 Genome Page. URL: http://cmr.jcvi.org/cgi-bin/CMR/GenomePage.cgi?org=ntbs01 (date of access: 15.04.2011).
  17. Iomantas Yurgis A. V., Abalkina E. G., Polanuer B. M., et al. Method for producing shikimic acid. US6436664.2002-08-20.
  18. Bron S. Plasmids. Molecular Biological Methods for Bacillus. Chichester : John Wiley a. Sons Ltd., 1990. P. 75–174.
  19. Vagner V., Dervyn E., Ehrlich S. D. A vector for systematic gene inactivation in Bacillus subtilis. J. Microbiol. 1998. Vol. 144, No. 11. P. 3097–3104.
Published
2018-05-02
Keywords: transformation, integration, inactivation, shikimic acid, shikimatkinase, PCR-screening, Bacillus subtilis
How to Cite
Chao, Y., Shonina, M. Y., & Lahodzich, A. V. (2018). Design of the general concept of regulated inactivation of the gene shikimatkinase for modification of the shikimat way in bacteria Bacillus subtilis. Experimental Biology and Biotechnology, 3, 45-53. Retrieved from https://journals.bsu.by/index.php/biology/article/view/2460
Section
Genetics and Molecular Biology