Co-production of 1,3-Propanediol and 2,3-Butanediol from Waste Lard by Co-cultivation of Pseudomonas alcaligenes and Klebsiella pneumoniae

  • Ying Li
  • Siyu Zhu
  • Xizhen GeEmail author


The platform chemicals 1,3-propanediol (1,3-PD) and 2,3-butanediol (2,3-BD) are important raw materials for polyesters and biofuels. However, the biosynthesis of the compounds relies on massive consumption of glucose or glycerol, leading to the uneconomical production in industrial scale. In this work, we developed a new method for co-production of 1,3-PD and 2,3-BD from waste lard to reduce the cost in carbon source supply. A waste lard utilizing Pseudomonas alcaligenes PA-3 and a 1,3-PD producing Klebsiella pneumoniae AA405 were co-cultivated by using waste lard as the sole carbon source. In a shake flask, 1.05 g/L 1,3-PD and 0.35 g/L 2,3-BD were produced from waste lard within 24 h. The addition of nitrogen source significantly increased the relative ratio of K. pneumoniae AA405 in the medium, which further favored to the higher titers of the two products. In bioreactor, the co-cultivation system produced 5.98 g/L 1,3-PD and 4.29 g/L 2,3-BD from 100 g/L waste lard within 72 h, and the conversion rate of 1,3-PD and 2,3-BD from waste lard were 62.95% and 0.75%, respectively. In all, this is the first work on 1,3-PD and 2,3-BD production from waste triglyceride, which will favor the utilization of low-cost carbon source in industrial production of chemicals.



This work was financially supported by Joint Funding of Beijing Municipal Natural Science Foundation-Beijing Municipal Education Commission, National Key R&D Program of China [2017YFD0201105] and Premium Funding Project for Academic Human Resources Development in Beijing Union University [BPHR2017DZ07].


  1. 1.
    Sun X, Shen X, Jain R, Lin Y, Wang J, Sun J, Wang J, Yan Y, Yuan Q (2015) Synthesis of chemicals by metabolic engineering of microbes. Chem Soc Rev 44:3760–3785CrossRefGoogle Scholar
  2. 2.
    Laursen JB, Nielsen J (2004) Phenazine natural products: biosynthesis, synthetic analogues, and biological activity. Chem Rev 104:1663–1686CrossRefGoogle Scholar
  3. 3.
    Maervoet VET, Maeseneire SLD, Avci FG, Beauprez J, Soetaert WK, Mey MD (2016) High yield 1,3-propanediol production by rational engineering of the 3-hydroxypropionaldehyde bottleneck in Citrobacter werkmanii. Microbial Cell Fact 15:23CrossRefGoogle Scholar
  4. 4.
    Jeong D, Yang J, Lee S, Kim B, Um Y, Kim Y, Ha KS, Lee J (2016) Deletion of the budBAC operon in Klebsiella pneumoniae to understand the physiological role of 2,3-butanediol biosynthesis. Prep Biochem 46:410–419CrossRefGoogle Scholar
  5. 5.
    Yang F, Hanna MA, Sun R (2012) Value-added uses for crude glycerol-a byproduct of biodiesel production. Biotechnol Biofuels 5:13CrossRefGoogle Scholar
  6. 6.
    Ma F, Hanna MA (1999) Biodiesel production: a review. Bioresour Technol 70:1–15CrossRefGoogle Scholar
  7. 7.
    Galadima A, Muraza O, Bundschuh J, Yusaf T, Maity JP, Nelson E, Mamat R, Mahlia TMI (2014) Biodiesel production from algae by using heterogeneous catalysts: a critical review. Energy 78:72–83CrossRefGoogle Scholar
  8. 8.
    Dias JM, Alvim-Ferraz MC, Almeida MF (2009) Production of biodiesel from acid waste lard. Bioresour Technol 100:6355–6361CrossRefGoogle Scholar
  9. 9.
    Adewale P, Dumont MJ, Ngadi M (2016) Enzyme-catalyzed synthesis and kinetics of ultrasonic assisted methanolysis of waste lard for biodiesel production. Chem Eng J 284:158–165CrossRefGoogle Scholar
  10. 10.
    Banković-Ilić IB, Stojković IJ, Stamenković OS, Veljkovic VB, Hung Y-T (2014) Waste animal fats as feedstocks for biodiesel production. Renew Sust Energ Rev 32:238–254CrossRefGoogle Scholar
  11. 11.
    Maggio-Hall LA, Keller NP (2010) Mitochondrial β-oxidation in Aspergillus nidulans. Mol Microbiol 54:1173–1185CrossRefGoogle Scholar
  12. 12.
    Piekarska K, Mol E, Berg MVD, Hardy G, Burg JVD, Roermund CV, Maccallum D, Odds F, Distel B (2006) Peroxisomal fatty acid β-oxidation is not essential for virulence of Candida albicans. Eukaryot Cell 5:1847–1856CrossRefGoogle Scholar
  13. 13.
    Marsudi S, Unno H, Hori K (2002) Palm oil utilization for the simultaneous production of polyhydroxyalkanoates and rhamnolipids by Pseudomonas aeruginosa. Biotechnol Bioeng 78:699–707CrossRefGoogle Scholar
  14. 14.
    Rahman KS, Thahira-Rahman J, Lakshmanaperumalsamy P, Banat IM (2002) Towards efficient crude oil degradation by a mixed bacterial consortium. Bioresour Technol 85:257–261CrossRefGoogle Scholar
  15. 15.
    Dwivedi P, Vivekanand V, Pareek N, Sharma A, Singh RP (2011) Co-cultivation of mutant E-3.510 and Pleurotus ostreatus for simultaneous biosynthesis of xylanase and laccase under solid-state fermentation. New Biotechnol 28:616–626CrossRefGoogle Scholar
  16. 16.
    Zhang H, Wang X (2016) Modular co-culture engineering, a new approach for metabolic engineering. Metab Eng 37:114–121CrossRefGoogle Scholar
  17. 17.
    Behçet R (2011) Performance and emission study of waste anchovy fish biodiesel in a diesel engine. Fuel Process Technol 92:1187–1194CrossRefGoogle Scholar
  18. 18.
    Johnson LA, Beacham IR, Macrae IC, Free ML (1992) Degradation of triglycerides by a pseudomonad isolated from milk: molecular analysis of a lipase-encoding gene and its expression in Escherichia coli. Appl Environ Microb 58:1776–1779Google Scholar
  19. 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 50:24–29CrossRefGoogle Scholar
  20. 20.
    Petrov K, Petrova P (2009) High production of 2,3-butanediol from glycerol by Klebsiella pneumoniae G31. Appl Microbiol Biotechnol 84:659–665CrossRefGoogle Scholar
  21. 21.
    Park JM, Rathnasingh C, Song H (2016) Metabolic engineering of Klebsiella pneumoniae based on in silico analysis and its pilot-scale application for 1,3-propanediol and 2,3-butanediol co-production. J Ind Microbiol Biotechnol 44:1–11Google Scholar
  22. 22.
    Metsoviti M, Paraskevaidi K, Koutinas A, Zeng AP, Papanikolaoua S (2012) 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 47:1872–1882CrossRefGoogle Scholar
  23. 23.
    Wang J, Zhao P, Li Y, Xu L, Tian P (2018) Engineering CRISPR interference system in Klebsiella pneumoniae for attenuating lactic acid synthesis. Microbial Cell Fact 17:56CrossRefGoogle Scholar
  24. 24.
    Li Y, Yin YM, Wang XY, Wu H, Ge XZ (2017) Evaluation of berberine as a natural fungicide: biodegradation and antimicrobial mechanism. J Asian Nat Prod Res 20:148–162CrossRefGoogle Scholar
  25. 25.
    Zelles L, Bai QY (1993) Fractionation of fatty acids derived from soil lipids by solid phase extraction and their quantitative analysis by GC-MS. Soil Biol Biochem 25:495–507CrossRefGoogle Scholar
  26. 26.
    Chen YM, Xiao B, Chang J, Fu Y, Lv PM, Wang XW (2009) Synthesis of biodiesel from waste cooking oil using immobilized lipase in fixed bed reactor. Energy Convers Manag 50:668–673CrossRefGoogle Scholar
  27. 27.
    Köpke M, Mihalcea C, Liew FM, Tizard JH, Ali MS, Conolly JJ, Alsinawi B, Simpson SD (2011) 2,3-butanediol production by acetogenic bacteria, an alternative route to chemical synthesis, using industrial waste gas. Appl Environ Microbiol 77:5467–5475CrossRefGoogle Scholar
  28. 28.
    Ying L, Xi W, Ge X, Tian P (2016) High production of 3-hydroxypropionic acid in Klebsiella pneumoniae by systematic optimization of glycerol metabolism. Sci Rep 6:26932CrossRefGoogle Scholar
  29. 29.
    Biebl H, Menzel K, Zeng AP, Deckwer WD (1999) Microbial production of 1,3-propanediol. Appl Microbiol Biotechnol 52:289–297CrossRefGoogle Scholar
  30. 30.
    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 20:82–86CrossRefGoogle Scholar
  31. 31.
    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 bio-diesel (fatty acid methyl esters) manufacturing process: production of 1,3-propanediol, citric acid and single cell oil. Biomass Bioenergy 32:60–71CrossRefGoogle Scholar
  32. 32.
    Zheng P, Wereath K, Sun J, Heuvel JVD, Zeng AP (2006) Overexpression of genes of the dha regulon and its effects on cell growth, glycerol fermentation to 1,3-propanediol and plasmid stability in Klebsiella pneumoniae. Process Biochem 41:2160–2169CrossRefGoogle Scholar
  33. 33.
    Zheng ZM, Cheng KK, Hu QL, Liu HJ, Guo NN, Liu DH (2008) Effect of culture conditions on 3-hydroxypropionaldehyde detoxification in 1,3-propanediol fermentation by Klebsiella pneumoniae. Biochem Eng J 39:305–310CrossRefGoogle Scholar
  34. 34.
    Chen Z, Liu H, Liu D (2009) Regulation of 3-hydroxypropionaldehyde accumulation in Klebsiella pneumoniae by overexpression of dhaT and dhaD genes. Enzyme Microb Technol 45:305–309CrossRefGoogle Scholar
  35. 35.
    Xiu ZL, Sun CYQ (2007) Stoichiometric analysis and experimental investigation of glycerol-glucose co-fermentation in Klebsiella pneumoniae under microaerobic conditions. Biochem Eng J 33:42–52CrossRefGoogle Scholar
  36. 36.
    Ji XJ, Huang H, Ouyang PK (2011) Microbial 2,3-butanediol production: a state-of-the-art review. Biotechnol Adv 29:351–364CrossRefGoogle Scholar
  37. 37.
    Yu EKC, Saddler JN (1982) Enhanced production of 2,3-butanediol by Klebsiella pneumoniae grown on high sugar concentrations in the presence of acetic acid. Appl Environ Microbiol 44:777–784Google Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Beijing Key Laboratory of Biomass Waste Resource Utilization, Biochemical Engineering CollegeBeijing Union UniversityBeijingChina

Personalised recommendations