Skip to main content
SpringerLink
Log in

A feeding strategy for tetramethylpyrazine production by Bacillus subtilis based on the stimulating effect of ammonium phosphate

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

To examine the effects of ammonium salts on tetramethylpyrazine (TTMP) production by Bacillus subtilis CCTCC M 208157, different ammonium salts were tested, and diammonium phosphate (DAP) was found to have a predominant effect on stimulating TTMP synthesis. The DAP requirements for TTMP production were then investigated, experimental results showed that higher concentrations of DAP favored TTMP production, while both the ammonium and phosphate ions exhibited inhibitory effects on the cell growth and precursor 3-hydroxy-2-butanone accumulation. Based on the results above, a DAP feeding strategy was developed and verified in further experiments. By applying the proposed fed-batch strategy, the maximum TTMP concentrations reached 7.46 and 7.34 g/l in flask and fermenter experiments, increased by 55.1 and 29.0% compared to that of the batch TTMP fermentation, respectively. To our knowledge, these results, i.e., TTMP yields in flask or fermenter fermentations, were new records on TTMP fermentation by B. subtilis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

TTMP:

2,3,5,6-Tetramethylpyrazine concentration (g/l)

HB:

3-Hydroxy-2-butanone concentration (g/l)

X :

Cell dry weight (g/l)

P :

Product concentration (g/l)

Q :

Product formation rate (g/l/h)

Y HB/X :

Yield of HB from biomass (g of HB/g of biomass)

Y TTMP/HB :

Yield of TTMP from HB (g of TTMP/g of HB)

References

  1. Seitz EW (1994) In: Gabelman A (ed) Bioprocess production of flavor, fragrance, and color ingredients. Wiley, New York

    Google Scholar 

  2. Masuda H, Mihara S (1988) Olfactive properties of alkylpyrazines and 3-substituted 2-alkylpyrazines. J Agric Food Chem 36:584–587

    Article  CAS  Google Scholar 

  3. Ho JW, Jie M (2007) Pharmacological activity of cardiovascular agents from herbal medicine. Cardiovasc Hematol Agents Med Chem 5:273–277

    Article  CAS  Google Scholar 

  4. Schrader J (2007) In: Berger RG (ed) Flavours and fragrances: chemistry, bioprocessing and sustainability. Springer, Berlin

    Google Scholar 

  5. Xiao ZJ, Xie NZ, Liu PH, Hua DL, Xu P (2006) Tetramethylpyrazine production from glucose by a newly isolated Bacillus mutant. Appl Microbiol Biotechnol 73:512–518

    Article  CAS  Google Scholar 

  6. Larroche C, Besson I, Gros JB (1999) High pyrazine production by Bacillus subtilis in solid substrate fermentation on ground soybeans. Process Biochem 34:667–674

    Article  CAS  Google Scholar 

  7. Besson I, Creuly C, Gros JB, Larroche C (1997) Pyrazine production by Bacillus subtilis in solid-state fermentation on soybeans. Appl Microbiol Biotechnol 47:489–495

    Article  CAS  Google Scholar 

  8. Demain AL, Jackson M, Trenner NR (1967) Thiamine-dependent accumulation of tetramethylpyrazine accompanying a mutation in isoleucine-valine pathway. J Bacteriol 94:323–326

    Article  CAS  Google Scholar 

  9. Kim KS, Lee HJ, Shon DH, Chung DK (1994) Optimum conditions for the production of tetramethylpyrazine flavor compound by aerobic fed-batch culture of Lactococcus lactis subsp. lactis biovar. diacetylactis FC1. J Microbiol Biotechnol 4:327–332

    CAS  Google Scholar 

  10. Zhu BF, Xu Y, Fan WL (2010) High-yield fermentative preparation of tetramethylpyrazine using an endogenous precursor approach by Bacillus sp. J Ind Microbiol Biotechnol 37:179–186

    Article  CAS  Google Scholar 

  11. Fan WL, Xu Y, Zhang YH (2007) Characterization of pyrazines in some Chinese liquors and their approximate concentrations. J Agric Food Chem 55:9956–9962

    Article  CAS  Google Scholar 

  12. Ji XJ, Huang H, Du J, Zhu JG, Ren LJ, Hu N et al (2009) Enhanced 2,3-butanediol production by Klebsiella oxytoca using a two-stage agitation speed control strategy. Bioresour Technol 100:3410–3414

    Article  CAS  Google Scholar 

  13. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  14. Maestri O, Joset F (2000) Regulation by external pH and stationary growth phase of the acetolactate synthase from Synechocystis PCC6803. Mol Microbiol 37:828–838

    Article  CAS  Google Scholar 

  15. Grundy FJ, Turinsky AJ, Henkin TM (1994) Catabolite regulation of Bacillus subtilis acetate and acetoin utilization genes by CcpA. J Bacteriol 176:4527–4533

    CAS  Google Scholar 

  16. Huang TC (1997) Combined effects of a buffer and solvent on tetramethylpyrazine formation in a 3-hydroxy-2-butanone/ammonium hydroxide system. Biosci Biotechnol Biochem 61:1013–1015

    Article  CAS  Google Scholar 

  17. Chung J, Shim H, Park SJ, Kim SJ, Bae W (2006) Optimization of free ammonia concentration for nitrite accumulation in shortcut biological nitrogen removal process. Bioprocess Biosyst Eng 28:275–282

    Article  CAS  Google Scholar 

  18. Nakashimada Y, Kanai K, Nishio N (1998) Optimization of dilution rate, pH and oxygen supply on optical purity of 2,3-butanediol produced by Paenibacillus polymyxa in chemostat culture. Biotechnol Lett 20:1133–1138

    Article  CAS  Google Scholar 

  19. Adachi T, Kamiya H, Kosuge T (1964) Studies on the metabolic products of Bacillus subtilis. IV. Determination and mechanism of formation of tetramethylpyrazine. Yakugaku Zasshi 84:545–548

    CAS  Google Scholar 

  20. Rizzi GP (1988) Formation of pyrazines from acyloin precursors under mild conditions. J Agric Food Chem 36:349–352

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This research was financially supported by National Key Technology R&D Program (2007BAK36B02 and 2008BAI63B06), the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) (No. IRT0532), the Program of Introducing Talents of Discipline to Universities (111 Project) (111-2-6) and the State Key Laboratory of Food Science and Technology, Jiangnan University (No. SKLF-MB-200801).

Author information

Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China

    Bing-Feng Zhu & Yan Xu

Authors
  1. Bing-Feng Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yan Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yan Xu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhu, BF., Xu, Y. A feeding strategy for tetramethylpyrazine production by Bacillus subtilis based on the stimulating effect of ammonium phosphate. Bioprocess Biosyst Eng 33, 953–959 (2010). https://doi.org/10.1007/s00449-010-0419-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-010-0419-5

Keywords

Search

Navigation