Abstract
3-Hydroxypropionaldehyde (3-HPA) is a toxic intermediary metabolite in the biological route of 1,3-propanediol biosynthesis from glycerol. 3-HPA accumulated in culture medium would arouse an irreversible cessation of the fermentation process. The role of substrate (glycerol) on 3-HPA accumulation in aerobic fermentation was investigated in this paper. 1,3-Propanediol oxidoreductase and glycerol dehydratase, two key enzyme catalyzing reactions of 3-HPA formation and consumption, were sensitive to high concentration of 3-HPA. When the concentration of 3-HPA increased to a higher level in medium (ac 10 mmol/L), the activity of 1,3-propanediol oxidoreductase in cell decreased correspondingly, which led to decrease of the 3-HPA conversion rate, then the 3-HPA concentration increasing was accelerated furthermore. 3-HPA accumulation in culture medium was triggered by this positive feedback mechanism. In the cell exponential growth phase, the reaction catalyzed by 1,3-propanediol oxidoreductase was the rate limiting step in 1,3-propanediol production. The level of 3-HPA in culture medium could be controlled by the substrate (glycerol) concentration, and lower level of glycerol could avoid 3-HPA accumulating to a high, lethal concentration. In fed batch fermentation, under the condition of initial glycerol concentration 30 g/L, and keeping glycerol concentration lower than 7–8 g/L in cell exponential growth phase, 3-HPA accumulation could not be incurred. Based on this result, a glycerol feeding strategy was set up in fed batch fermentation. Under the optimized condition, 50.1 g/L of 1,3-propanediol was produced in 24 h, and 73.1 g/L of final 1,3-propanediol concentration was obtained in 54 h.
Similar content being viewed by others
References
Abbad-Andaloussi S, Guedon E, Spiesser E, Petitdemange H (1996) Glycerol dehydratase activity: the limiting step for 1,3-propanediol production by Clostridium butyricum. Lett Appl Microbiol 22:311–314. doi:10.1111/j.1472-765X.1996.tb01168.x
Abbad-Andaloussi S, Manginot-Durr C, Amine J, Petitdemange E, Petitdemange H (1995) Isolation and characterisation of Clostridium butyricum DSM 5431 mutants with increased resitance to 1,3-propanediol and altered production of acids. Appl Environ Microbiol 61:4413–4417
Ahrens K, Menzel K, Zeng AP, Deckwer WD (1998) Kinetic, dynamic, and pathway studies of glycerol metabolism by Klebsiella pneumoniae in anaerobic continuous culture. Part III: enzymes and fluxes of glycerol dissimilation and 1,3-propanediol formation. Biotechnol Bioeng 59:544–552. doi :10.1002/(SICI)1097-0290(19980905)59:5<544::AID-BIT3>3.0.CO;2-A
Arntz D, Haas T, Müller A, Wiegand N (1991) Kinetische untersuchung zur hydratisierung von acrolein. Chem Ing Tech 63:733–735. doi:10.1002/cite.330630714
Barbirato F, Grivet JP, Soucaille P, Bories A (1996) 3-Hydroxypropionaldehyde, an inhibitory metabolite of glycerol fermentation to 1,3-propanediol by enterobacterial species. Appl Environ Microbiol 62:1448–1451
Barbirato F, Soucaille P, Bories A (1996) Physiologic mechanisms involved in accumulation of 3-hydroxypropionaldehyde during fermentation of glycerol by Enterobacter agglomerans. Appl Environ Microbiol 62(12):4405–4409
Biebl H (2001) Fermentation of glycerol by Clostridium pasteurianum—batch and continuous culture studies. J Ind Microbiol Biotechnol 27:18–26. doi:10.1038/sj.jim.7000155
Boenigk R, Bowien S, Gottschalk G (1993) Fermentation of glycerol to 1,3-propanediol in continuous cultures of Citrobacter freundii. Appl Microbiol Biotechnol 38:453–457. doi:10.1007/BF00242936
Cameron DC, Altaras NE, Hoffman ML, Shaw AJ (1998) Metabolic engineering of propanediol pathways. Biotechnol Prog 14:116–125. doi:10.1021/bp9701325
Cirde SJ, Stone L, Boruff CS (1945) Acrolein determination by means of tryptophane. Ind Eng Chem Anal Ed 17:259–262. doi:10.1021/i560140a021
Decker WD (1995) Microbial conversion of glycerol to 1,3-propanediol. FEMS Microbiol Rev 16:143–149. doi:10.1111/j.1574-6976.1995.tb00162.x
Forage RG, Lin ECC (1982) dha systems mediating aerobic and anaerobic dissimilation of glycerol in Klebsiella pneumoniae NCIB 418. J Bacteriol 151:591–599
Johnson EA, Lin ECC (1987) Klebsiella pneumoniae 1,3-propanediol: NAD + oxidoreductase. J Bacteriol 169:2050–2054
Hao J, Lin RH, Zheng ZM, Liu HJ, Liu DH (2008) Isolation and characterization of microorganisms able to produce 1,3-propanediolunder aerobic conditions. World J Microbiol Biotechnol 24:1731–1740. doi:10.1007/s11274-008-9665-y
Hao J, Wang W, Tian JS, Li JL, Liu DH (2008) Decrease of 3-hydroxypropionaldehyde accumulation in 1,3-propanediol production by over-expressing dhaT gene in Klebsiella pneumoniae TUAC01. J Ind Microbiol Biotechnol 35:735–741. doi:10.1007/s10295-008-0340-y
Homann T, Tag C, Biebl H, Deckwer WD, Schink B (1990) Fermentation of glycerol to 1,3-propanediol by Klebsiella and Citrobacter strains. Appl Microbiol Biotechnol 33:121–126. doi:10.1007/BF00176511
Knifton JF, James TG, Slaugh LH, Allen KD, Weider PR, Powell JB (2004) One-step production of 1,3-propanediol from ethylene oxide and syngas with a cobalt–iron catalyst. US Patent. 6.750.373
Liu HJ, Zhang DJ, Xu YH, Mu Y, Sun YQ, Xiu ZL (2007) Microbial production of 1,3-propanediol from glycerol by Klebsiella pneumoniae under micro-aerobic conditions up to a pilot scale. Biotechnol Lett 29:1281–1285. doi:10.1007/s10529-007-9398-2
Menzel K, Zeng AP, Deckwer WD (1997) High concentration and productivity of 1,3-propanediol from continuous fermentation of glycerol by Klebsiella pneumoniae. Enzyme Microb Technol 20:82–86. doi:10.1016/S0141-0229(96)00087-7
Menzel K, Ahrens K, Zeng AP, Deckwer WD (1998) Kinetic, dynamic, and pathway studies of glycerol metabolism by Klebsiella pneumoniae in anaerobic continuous culture: IV. Enzymes and fluxes of pyruvate metabolism. Biotechnol Bioeng 60:617–626. doi:10.1002/(SICI)1097-0290(19981205)60:5<617::AID-BIT12>3.0.CO;2-L
Nakamura CE, Whitedy GM (2003) Metabolic engineering for the microbial production of 1,3-propanediol. Curr Opin Biotechnol 14:454–459. doi:10.1016/j.copbio.2003.08.005
Papanikolaoua S, Fakas S, Ficka M, Chevalota I, Galiotou-Panayotoub M, Komaitisb M et al (2008) Biotechnological valorisation of raw glycerol discharged after bio-diesel (fatty acid methyl esters) manufacturing process: production of 1,3-propanediol, citric acid and single cell oil. Biomass Bioenergy 32:60–71. doi:10.1016/j.biombioe.2007.06.007
Papanikolaou S, Patricia RS, Bernard P, Blancharda F, Fick M (2000) High production of 1,3-propanediol from industrial glycerol by a newly isolated Clostridium butyricum strain. J Biotechnol 77:191–208. doi:10.1016/S0168-1656(99)00217-5
Pflugmacher U, Gottschalk G (1994) Development of an immobilized cell reactor for the production of 1,3-propanediol by Citrobacter freundii. Appl Microbiol Biotechnol 41:313–316. doi:10.1007/BF00221225
Reimann A, Biebl H (1996) Production of 1,3-propanediol by Clostridium butyricum DSM 5431 and product tolerant mutants in fedbatch culture: feeding strategy for glycerol and ammonium. Biotechnol Lett 18:827–832. doi:10.1007/BF00127897
Reimann A, Biebl H, Deckwer WD (1998) Production of 1,3-propanediol by Clostridium butyricum in continuous culture with cell recycling. Appl Microbiol Biotechnol 49:359–363. doi:10.1007/s002530051182
Yang G, Tian JS, Li JL (2007) Fermentation of 1,3-propanediol by a lactate deficient mutant of Klebsiella oxytoca under microaerobic conditions. Appl Microbiol Biotechnol 73:1017–1024. doi:10.1007/s00253-006-0563-7
Zeng AP, Bieb H (2002) Bulk chemicals from biotechnology: the case of 1,3-propanediol production and the new trends. Adv Biochem Eng Biotechnol 74:239–259
Zhao YN, Chen G, Yao SJ (2006) Microbial production of 1,3-propanediol from glycerol by encapsulated Klebsiella pneumoniae. Biochem Eng J 32:93–99. doi:10.1016/j.bej.2006.09.007
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Hao, J., Lin, R., Zheng, Z. et al. 3-Hydroxypropionaldehyde guided glycerol feeding strategy in aerobic 1,3-propanediol production by Klebsiella pneumoniae . J Ind Microbiol Biotechnol 35, 1615–1624 (2008). https://doi.org/10.1007/s10295-008-0405-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10295-008-0405-y