Applied Microbiology and Biotechnology

, Volume 94, Issue 3, pp 651–658 | Cite as

Enhanced production of 2,3-butanediol by engineered Bacillus subtilis

  • Ranjita Biswas
  • Masaru Yamaoka
  • Hideki Nakayama
  • Takashi Kondo
  • Ken-ichi Yoshida
  • Virendra S. Bisaria
  • Akihiko Kondo
Biotechnological products and process engineering

Abstract

Production of 2,3-butanediol by Bacillus subtilis takes place in late-log or stationary phase, depending on the expression of bdhA gene encoding acetoin reductase, which converts acetoin to 2,3-butanediol. The present work focuses on the development of a strain of B. subtilis for enhanced production of 2,3-butanediol in early log phase of growth cycle. For this, the bdhA gene was expressed under the control of PalsSD promoter of AlsSD operon for acetoin fermentation which served the substrate for 2,3-butanediol production. Addition of acetic acid in the medium induced the production of 2,3-butanediol by 2-fold. Two-step aerobic–anaerobic fermentation further enhanced 2,3-butanediol production by 4-fold in comparison to the control parental strain. Thus, addition of acetic acid and low dissolved oxygen in the medium are involved in activation of bdhA gene expression from PalsSD promoter in early log phase. Under the conditions tested in this work, the maximum production of 2,3-butanediol, 2.1 g/l from 10 g/l glucose, was obtained at 24 h. Furthermore, under the optimized microaerophilic condition, the production of 2,3-butanediol improved up to 6.1 g/l and overall productivity increased by 6.7-fold to 0.4 g/l h in the engineered strain compared to that in the parental control.

Keywords

2,3-Butanediol Bacillus subtilis Anaerobic fermentation bdhA gene Microaerophilic 

References

  1. Celinska E, Grajek W (2009) Biotechnological production of 2,3-butanediol—current state and prospects. Biotechnol Adv 27:715–725CrossRefGoogle Scholar
  2. de Boer AS, Diderichsen B (1991) On the safety of Bacillus subtilis and B. amyloliquefaciens: a review. Appl Microbiol Biotechnol 36:1–4CrossRefGoogle Scholar
  3. Hespell RB (1996) Fermentation of xylan, corn fiber, or sugars to acetoin and butanediol by Bacillus polymyxa strains. Curr Microbiol 32:291–296CrossRefGoogle Scholar
  4. Holtzclaw WD, Chapman LF (1975) Degradative acetolactate synthase of Bacillus subtilis: purification and properties. J Bacteriol 121:917–922Google Scholar
  5. Jansen NB, Flickinger MC, Tsao GT (1984) Production of 2,3-butanediol from d-xylose by Klebsiellaoxytoca ATCC 8724. Biotechnol Bioeng 26:362–369CrossRefGoogle Scholar
  6. Ji XJ, Huang H, Du J, Zhu JG, Ren LJ, Li S, Nie ZK (2009) Development of an industrial medium for economical 2,3-butanediol production through co-fermentation of glucose and xylose by Klebsiellaoxytoca. Bioresour Technol 100:5214–5218CrossRefGoogle Scholar
  7. Juni E (1952) The mechanism of formation of acetoin by bacteria. J Biol Chem 195:715–726Google Scholar
  8. Kunst F, Ogasawara N, Moszer I, Albertini AM, Alloni G, Azevedo V, Bertero MG, Bessières P, Bolotin A, Borchert S, Borriss R, Boursier L, Brans A, Braun M, Brignell SC, Bron S, Brouillet S, Bruschi CV, Caldwell B, Capuano V, Carter NM, Choi SK, Codani JJ, Connerton IF, Cummings NJ, Daniel RA, Denizot F, Devine KM, Düsterhöft A, Ehrlich SD, Emmerson PT, Entian KD, Errington J, Fabret C, Ferrari E, Foulger D, Fritz C, Fujita M, Fujita Y, Fuma S, Galizzi A, Galleron N, Ghim SY, Glaser P, Goffeau A, Golightly EJ, Grandi G, Guiseppi G, Guy BJ, Haga K, Haiech J, Harwood CR, Hénaut A, Hilbert H, Holsappel S, Hosono S, Hullo MF, Itaya M, Jones L, Joris B, Karamata D, Kasahara Y, Klaerr-Blanchard M, Klein C, Kobayashi Y, Koetter P, Koningstein G, Krogh S, Kumano M, Kurita K, Lapidus A, Lardinois S, Lauber J, Lazarevic V, Lee SM, Levine A, Liu H, Masuda S, Mauël C, Médigue C, Medina N, Mellado RP, Mizuno M, Moest D, Nakai S, Noback M, Noone D, O’Reilly M, Ogawa K, Ogiwara A, Oudega B, Park SH, Parro V, Pohl TM, Portetelle D, Porwollik S, Prescott AM, Presecan E, Pujic P, Purnelle B, Rapoport G, Rey M, Reynolds S, Rieger M, Rivolta C, Rocha E, Roche B, Rose M, Sadaie Y, Sato T, Scanlan E, Schleich S, Schroeter R, Scoffone F, Sekiguchi J, Sekowska A, Seror SJ, Serror P, Shin BS, Soldo B, Sorokin A, Tacconi E, Takagi T, Takahashi H, Takemaru K, Takeuchi M, Tamakoshi A, Tanaka T, Terpstra P, Tognoni A, Tosato V, Uchiyama S, Vandenbol M, Vannier F, Vassarotti A, Viari A, Wambutt R, Wedler E, Wedler H, Weitzenegger T, Winters P, Wipat A, Yamamoto H, Yamane K, Yasumoto K, Yata K, Yoshida K, Yoshikawa HF, Zumstein E, Yoshikawa H, Danchin A (1997) The complete genome sequence of the Gram-positive bacterium Bacillus subtilis. Nature 390:249–256CrossRefGoogle Scholar
  9. Li D, Dai JY, Xiu ZL (2010a) A novel strategy for integrated utilization of Jerusalem artichoke stalk and tuber for production of 2,3-butanediol by Klebsiella pneumoniae. Bioresour Technol 101:8342–8347CrossRefGoogle Scholar
  10. Li ZJ, Jian J, Wei XX, Shen XW, Chen GQ (2010b) Microbial production of meso-2,3-butanediol by metabolically engineered Escherichia coli under low oxygen condition. Appl Microbiol Biotechnol 87:2001–2009CrossRefGoogle Scholar
  11. Morinaga T, Ashida H, Yoshida K (2010) Identification of two scyllo-inositol dehydrogenases in Bacillus subtilis. Microbiology 156:1538–1546CrossRefGoogle Scholar
  12. Nakashimada Y, Marwoto B, Kashiwamura T, Kakizono T, Nishio N (2000) Enhanced 2,3-butanediol production by addition of acetic acid in Paenibacillus polymyxa. J Biosci Bioeng 90:661–664Google Scholar
  13. Nicholson WL (2008) The Bacillus subtilisy djL (bdhA) gene encodes acetoin reductase 2,3-butanediol dehydrogenase. Appl Environ Microbiol 74:6832–6838CrossRefGoogle Scholar
  14. Qureshi N, Cheryan M (1989) Effects of aeration on 2,3-butanediol production from glucose by Klebsiella oxytoca. J Ferment Bioeng 67:415–418CrossRefGoogle Scholar
  15. Ramos HC, Hoffmann T, Marino M, Nedjari H, Presecan-siedel E, Dreesen O, Glaserand P, Jahn D (2000) Fermentative metabolism of Bacillus subtilis: physiology and regulation of gene expression. J Bacteriol 182:3072–3080CrossRefGoogle Scholar
  16. Renna MC, Najimudin N, Winik LR, Zahler SA (1993) Regulation of the Bacillus subtilis alsS, alsD and alsR genes involved in the post-exponential-phase production of acetoin. J Bacteriol 175:3863–3875Google Scholar
  17. Sablayrolles JM, Goma G (1984) Butanediol production by Aerobacteraerogenes NRRL B199: effects of initial substrate concentration and aeration agitation. Biotechnol Bioeng 26:148–155CrossRefGoogle Scholar
  18. Schallmey M, Singh A, Ward OP (2004) Developments in the use of Bacillus species for industrial production. Can J Microbiol 50:1–17CrossRefGoogle Scholar
  19. Sonenshein AL (2000) Control of sporulation initiation in Bacillus subtilis. Curr Opin Microbiol 3:561–566CrossRefGoogle Scholar
  20. Stülke J, Hillen W (2000) Regulation of carbon catabolism in Bacillus species. Annu Rev Microbiol 54:849–880CrossRefGoogle Scholar
  21. Tsau JL, Guffanti AA, Montville TJ (1992) Conversion of pyruvate to acetoin helps to maintain pH homeostasis in Lactobacillus plantarum. Appl Environ Microbiol 58:891–894Google Scholar
  22. Westers L, Westers H, Quax WJ (2004) Bacillus subtilis as cell factory for pharmaceutical proteins: a biotechnological approach to optimize the host organism. Biochim Biophys Acta 1694:299–310CrossRefGoogle Scholar
  23. Yoshida K, Fujita Y, Ehrlich SD (1999) Three asparagines synthetase genes of Bacillus subtilis. J Bacteriol 181:6081–6091Google Scholar
  24. Yoshida K, Aoyama D, Ishio I, Shibayama T, Fujita Y (1997) Organization and transcription of the myo-inositol operon, iol, of Bacillus subtilis. J Bacteriol 179:4591–4598Google Scholar
  25. Yu EKC, Saddler JN (1982) Enhanced production of 2,3-butanediol by Klebsilla pneumonia grown on high sugar concentration in the presence of acetic acid. Appl Environ Microbiol 44:777–784Google Scholar
  26. Zeng AP, Sabra W (2011) Microbial production of diols as platform chemicals: recent progresses. Curr Opin Biotechnol. doi:10.1016/j.copbio.2011.05.005
  27. Zhang L, Yang Y, Sun J, Shen Y, Wei D, Zhu J, Chu J (2010) Microbial production of 2,3-butanediol by a mutagenized strain of Serratia marcescens H30. Bioresour Technol 101:1961–1967CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Ranjita Biswas
    • 1
  • Masaru Yamaoka
    • 2
  • Hideki Nakayama
    • 1
  • Takashi Kondo
    • 1
  • Ken-ichi Yoshida
    • 2
  • Virendra S. Bisaria
    • 3
  • Akihiko Kondo
    • 4
  1. 1.Organization of Advance Science and TechnologyKobe UniversityKobeJapan
  2. 2.Department of Agrobioscience, Graduate School of Agricultural ScienceKobe UniversityKobeJapan
  3. 3.Department of Biochemical Engineering and BiotechnologyIndian Institute of Technology DelhiNew DelhiIndia
  4. 4.Department of Chemical Science and Engineering, Graduate School of EngineeringKobe UniversityKobeJapan

Personalised recommendations