Abstract
Cultivation in glycerol instead of sugars inhibits 2,3-butanediol (2,3-BD) production by Bacillus amyloliquefaciens. In this study, we report that B. amyloliquefaciens readily produces 2,3-BD from biodiesel-derived glycerol in the presence of beet molasses as a co-substrate. Unexpectedly, the molasses stimulated 2,3-BD production and simultaneously reduced the duration of fermentation. Productivity of 2,3-BD was enhanced at the start of fermentation, and yields increased under continuous molasses supply. Subsequently, 2,3-BD production in molasses-supplemented fed-batch culture was observed. Prior to inoculation of fed-batch fermentation culture, 15 g/l of molasses was added to the bioreactor. After 6 h of incubation, the bioreactor was fed with a solution containing 80 % glycerol and 15 % molasses. The 2,3-BD concentration, yield, and productivity significantly improved, reaching 83.3 g/l, 0.42 g/g, and 0.87 g/l·h, respectively. To our knowledge, these results are the highest report for 2,3-BD fermentation from biodiesel-derived glycerol.
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References
Abbad-Andaloussi S, Amine J, Gerard P, Petitdemange H (1998) Effect of glucose on glycerol metabolism by Clostridium butyricum DSM 5431. J Appl Microbiol 84:515–522
Adhya S, Echols H (1966) Glucose effect and the galactose enzymes of Escherichia coli: correlation between glucose inhibition of induction and inducer transport. J Bacteriol 92(3):601–608
Biebl H, Marten S (1995) Fermentation of glycerol to 1,3-propanediol: use of cosubstrates. Appl Microbiol Biotechnol 44:15–19
Biebl H, Zeng AP, Menzel K, Deckwer WD (1998) Fermentation of glycerol to 1,3-propanediol and 2,3-butanediol by Klebsiella pneumoniae. Appl Microbiol Biotechnol 50(1):24–29
Blomqvist K, Nikkola M, Lehtovaara P, Suihko ML, Airaksinen U, Straby KB, Knowles JK, Penttila ME (1993) Characterization of the genes of the 2,3-butanediol operons from Klebsiella terrigena and Enterobacter aerogenes. J Bacteriol 175(5):1392–1404
Booth I (1985) Regulation of cytoplasmic pH in bacteria. Microbiol Rev 49(4):359–378
Celinska E, Grajek W (2009) Biotechnological production of 2,3-butanediol—current state and prospects. Biotechnol Adv 27(6):715–725
Da Silva GP, Mack M, Contiero J (2009) Glycerol: a promising and abundant carbon source for industrial microbiology. Biotechnol Ad 27(1):30–39
Ezeji TC, Qureshi N, Blaschek HP (2004) Acetone butanol ethanol (ABE) production from concentrated substrate: reduction in substrate inhibition by fed-batch technique and product inhibition by gas stripping. Appl Microbiol Biotechnol 63:653–655
Ji XJ, Huang H, Ouyang PK (2011) Microbial 2,3-butanediol production: a state-of-the-art review. Biotechnol Adv 29(3):351–364
Jiménez AM, Borja R, Martı́n A (2004) A comparative kinetic evaluation of the anaerobic digestion of untreated molasses and molasses previously fermented with Penicillium decumbens in batch reactors. Biochem Eng J 18(2):121–132
Johnson DT, Taconi KA (2007) The glycerin glut: options for the value-added conversion of crude glycerol resulting from biodiesel production. Environ Prog 26(4):338–348
Ma C, Wang A, Qin J, Li L, Ai X, Jiang T, Tang H, Xu P (2009) Enhanced 2,3-butanediol production by Klebsiella pneumoniae SDM. Appl Microbiol Biotechnol 82(1):49–57
Metsoviti M, Paramithiotis S, Drosinos EH, Galiotou-Panayotou M, Nychas G-JE, Zeng A-P, Papanikolaou S (2012a) Screening of bacterial strains capable of converting biodiesel-derived raw glycerol into 1,3-propanediol, 2,3-butanediol, and ethanol. Eng Life Sci 12(1):57–68
Metsoviti M, Paraskevaidi K, Koutinas A, Zeng A-P, Papanikolaou S (2012b) Production of 1,3-propanediol, 2,3-butanediol, and ethanol by a newly isolated Klebsiella oxytoca strain growing on biodiesel-derived glycerol based media. Process Biochem 47(12):1872–1882
Monod J (1942). Recherches sur la croissance des cultures bacteriennes. Paris, Hermann et Cie
Monod J (1947) The phenomenon of enzymatic adaptation. Growth 11:223–289
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(6):661–664
Papanikolaou S, Fakas S, Fick M, Chevalot I, Galiotou-Panayotou M, Komaitis M, Marc I, Aggelis G (2008) Biotechnological valorisation of raw glycerol discharged after biodiesel (fatty acid methyl esters) manufacturing process: production of 1,3-propanediol, citric acid, and single cell oil. Biomass Bioenergy 32(1):60–71
Petrov K, Petrova P (2009) High production of 2,3-butanediol from glycerol by Klebsiella pneumoniae G31. Appl Microbiol Biotechnol 84(4):659–665
Petrov K, Petrova P (2010) Enhanced production of 2,3-butanediol from glycerol by forced pH fluctuations. Appl Microbiol Biotechnol 87(3):943–949
Saint-Amans S, Girbal L, Andrade J, Ahrens K, Soucaille P (2001) Regulation of carbon and electron flow in Clostridium butyricum VPI 3266 grown on glucose–glycerol mixtures. J Bacteriol 183(5):1748–1754
Sattayasamitsathita S, Prasertsana P, Methacanon P (2011) Statistical optimization for simultaneous production of 1,3-propanediol and 2,3-butanediol using crude glycerol by newly bacterial isolate. Process Biochem 46:608–614
Syu MJ (2001) Biological production of 2,3-butanediol. Appl Microbiol Biotechnol 55(1):10–18
Yang G, Tian J, Li J (2007) Fermentation of 1,3-propanediol by a lactate deficient mutant of Klebsiella oxytoca under microaerobic conditions. Appl Microbiol Biotechnol 73(5):1017–1024
Yang T, Rao Z, Zhang X, Lin Q, Xia H, Xu Z, Yang S (2011) Production of 2,3-butanediol from glucose by GRAS microorganism Bacillus amyloliquefaciens. J Basic Microbiol 51(6):650–658
Yang T, Zhang X, Rao Z, Gu S, Xia H, Xu Z (2012) Optimization and scale-up of 2,3-butanediol production by Bacillus amyloliquefaciens B10-127. World J Microbiol Biotechnol 28:1563–1574
Zeng A-P, Sabra W (2011) Microbial production of diols as platform chemicals: recent progresses. Curr Opin Biotechnol 22(6):749–757
Acknowledgments
This work was supported by the Program for New Century Excellent Talents in University (NCET-10-0459), the National Basic Research Program of China (973 program) (2012CB725202), the High-tech Research and Development Programs of China (2011AA02A211, 2012AA022102), the National Natural Science Foundation of China (21276110), the Fundamental Research Funds for the Central Universities (JUSRP51306A, JUSRP1009), the research fund for the Doctoral Program of Higher Education of China (20110093120001), and a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Yang, TW., Rao, ZM., Zhang, X. et al. Fermentation of biodiesel-derived glycerol by Bacillus amyloliquefaciens: effects of co-substrates on 2,3-butanediol production. Appl Microbiol Biotechnol 97, 7651–7658 (2013). https://doi.org/10.1007/s00253-013-5048-x
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DOI: https://doi.org/10.1007/s00253-013-5048-x