Skip to main content
SpringerLink
Log in

Effects of immobilization of Actinobacillus succinogenes on efficiency of bio-succinic acid production from glycerol

  • Original Article
  • Published:
Biomass Conversion and Biorefinery Aims and scope Submit manuscript

Abstract

The aim of this research was to optimize bio-succinic acid production from glycerol by using Actinobacillus succinogenes. The effects of different substrate types (pure and crude glycerol), cell patterns (free cells and immobilized cells in agar), and carbonate sources (MgCO3 and CaCO3) on succinic (SA) production were studied in serum bottles. The maximum succinic acid concentration of 8.9 ± 0.5 g/L and yield of 1.27 g SA/g GLR were achieved using crude glycerol fermentation by immobilized cells supplemented with10 g/L of MgCO3 to increase dissolved CO2 in the system. However, a high concentration of MgCO3 (20 g/L) resulted in an adverse effect on SA production due to increasing pH which decreased CO2 fixation in the C4 pathway. The optimal conditions were used for up-scaling SA fermentation in a 1-L bioreactor using batch and semi-continuous operations. Higher SA concentration, glycerol utilization, and SA yield of 10.8 g/L, 88.8%w/w, and 1.25 g SA/g GLR, respectively, were achieved using the batch process. The development process is potent for further up-scaling the study for SA production.

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
Fig. 5

Similar content being viewed by others

References

  1. Singh D, Sharma D, Soni SL, Sharma S, Kumar SP, Jhalani A (2020) A review on feedstocks, production processes, and yield for different generations of biodiesel. Fuel 262:116553. https://doi.org/10.1016/j.fuel.2019.116553

    Article  Google Scholar 

  2. National Biodiesel Board (2019) Production Statistics. https://www.biodiesel.org/production/production-statistics. Accessed 30 Nov 2019

  3. Department of Energy Business (2019) Biodiesel production volume (B100) of Thailand. Bangkok: Department of Alternative Energy Development and Efficiency. http://www.dede.go.th/ewt_dl_link.php?nid = 362&filename = Biodiesel. Accessed 20 October 2019

  4. Quispe CAG, Coronado CJR, Carvalho JA Jr (2013) Glycerol: Production, consumption, prices, characterization and new trends in combustion. Renew Sust Energ Rev 27:475–493. https://doi.org/10.1016/j.rser.2013.06.017

    Article  Google Scholar 

  5. Luo X, Ge X, Cui S, Li Y (2016) Value-added processing of crude glycerol into chemicals and polymers. Bioresour Technol 215:144–154. https://doi.org/10.1016/j.biortech.2016.03.042

    Article  Google Scholar 

  6. Carvalho M, Matos M, Roca C, Reis MA (2014) Succinic acid production from glycerol by Actinobacillus succinogenes using dimethylsulfoxide as electron acceptor. New Biotechnol 31(1):133–139. https://doi.org/10.1016/j.nbt.2013.06.006

    Article  Google Scholar 

  7. Pinazo JM, Domine ME, Parvulescu V, Petru F (2015) Sustainability metrics for succinic acid production: a comparison between biomass-based and petrochemical routes. Catal Today 239:17–24. https://doi.org/10.1016/j.cattod.2014.05.035

    Article  Google Scholar 

  8. Saxena RK, Saran S, Isar J, Kaushik R (2017) Production and applications of succinic acid. In: Chen W, Goto M, Koutinas A (eds) Current Developments in Biotechnology and Bioengineering, 1st edn. Elsevier, Amsterdam, pp 601–630

    Chapter  Google Scholar 

  9. Lee CP, Lee WG, Lee SY, Chang HN (2010) Succinic acid production with reduced by-product formation in the fermentation of Anaerobiospirillum succiniciproducens using glycerol as a carbon source. Biotechnol Bioeng 41-48. https://doi.org/10.1002/1097-0290(20010105)72:1%3C41::AID-BIT6%3E3.0.CO;2-N

  10. Scholten E, Dägele D (2008) Succinic acid production by a newly isolated bacterium. Biotechnol Lett 2143-2146. https://doi.org/10.1007/s10529-008-9806-2

  11. Olajuyin AM, Yang M, Thygesen A, Tian J, Mu T, Xing J (2019) Effective production of succinic acid from coconut water (Cocos nucifera) by metabolically engineered Escherichia coli with overexpression of Bacillus subtilis pyruvate carboxylase. Biotechnol Rep 24:e00378. https://doi.org/10.1016/j.btre.2019.e00378

    Article  Google Scholar 

  12. Li J, Jiang M, Chen KQ, Ye Q, Shang LA, Wei P, Ying HJ, Chang HN (2010) Effect of redox potential regulation on succinic acid production by Actinobacillus succinogenes. Bioprocess Biosyst Eng 33(8):911–920. https://doi.org/10.1007/s00449-010-0414-x

    Article  Google Scholar 

  13. Vlysidis A, Binns M, Webb C, Theodoropoulos C (2011) Glycerol utilisation for the production of chemicals: Conversion to succinic acid, a combined experimental and computational study. Biochem Eng J 58-59:1–11. https://doi.org/10.1016/j.bej.2011.07.004

    Article  Google Scholar 

  14. Guettler MV, Mahendra K, Soni JK (1996) Process for making succinic acid, microorganisms for use in the process and methods of obtaining the microorganisms. United States Patent US OO5504004A

  15. Rigaki A, Webb C, Theodoropoulos C (2020) Double substrate limitation model for the bio-based production of succinic acid from glycerol. Biochem Eng J 153:107391. https://doi.org/10.1016/j.bej.2019.107391

    Article  Google Scholar 

  16. Corona-Gonzalez RI, Miramontes-Murillo R, Arriola-Guevara E, Guatemala-Morales G, Toriz G, Pelayo-Ortiz C (2014) Immobilization of Actinobacillus succinogenes by adhesion or entrapment for the production of succinic acid. Bioresour Technol 164:113–118. https://doi.org/10.1016/j.biortech.2014.04.081

    Article  Google Scholar 

  17. Najafpour GD (2015) Immobilization of microbial cells for the production of organic acid and ethanol. In: Chen W, Goto M, Koutinas A (eds) Biochemical Engineering and Biotechnology, 2nd edn. Elsevier, Amsterdam, pp 257–289

    Chapter  Google Scholar 

  18. Kikani BA, Pandey S, Singh SP (2013) Immobilization of the alpha-amylase of Bacillus amyloliquifaciens TSWK1-1 for the improved biocatalytic properties and solvent tolerance. Bioprocess Biosyst Eng 36(5):567–577. https://doi.org/10.1007/s00449-012-0812-3

    Article  Google Scholar 

  19. Song H, Lee JW, Choi S, You JK, Hong WH, Lee SY (2007) Effects of dissolved CO2 levels on the growth of Mannheimia succiniciproducens and succinic acid production. Biotechnol Bioeng 98(6):1296–1304. https://doi.org/10.1002/bit.21530

    Article  Google Scholar 

  20. Tan JP, Luthfi AAI, Manaf SFA, Wu TY, Jahim JM (2018) Incorporation of CO2 during the production of succinic acid from sustainable oil palm frond juice. J CO2 Util 26:595–601. https://doi.org/10.1016/j.jcou.2018.06.006

    Article  Google Scholar 

  21. Pateraki C, Patsalou M, Vlysidis A, Kopsahelis N, Webb C, Koutinas AA, Koutinas M (2016) Actinobacillus succinogenes: Advances on succinic acid production and prospects for development of integrated biorefineries. Biochem Eng J 112:285–303. https://doi.org/10.1016/j.bej.2016.04.005

    Article  Google Scholar 

  22. McKinlay JB, Laivenieks M, Schindler BD, McKinlay AA, Siddaramappa S, Challacombe JF et al (2010) A genomic perspective on the potential of Actinobacillus succinogenes for industrial succinate production. BMC Genomics 11:680. https://doi.org/10.1186/1471-2164-11-680

    Article  Google Scholar 

  23. Van der Werf MJ, Guettler MV, Jain MK, Zeikus JG (1997) Environmental and physiological factors affecting the succinate product ratio during carbohydrate fermentation by Actinobacillus sp. 130Z. Arch Microbiol 167:332–342. https://doi.org/10.1007/s002030050452

    Article  Google Scholar 

  24. Xi YL, Chen KQ, Li J, Fang XJ, Zheng XY, Sui SS, Jiang M, Wei P (2011) Optimization of culture conditions in CO2 fixation for succinic acid production using Actinobacillus succinogenes. J Ind Microbiol Biotechnol 38(9):1605–1612. https://doi.org/10.1007/s10295-011-0952-5

    Article  Google Scholar 

  25. Tanthunwet M, Tanthunwet M (2004) Chemistry of water and wastewater. Chulalongkorn University, Bangkok

    Google Scholar 

  26. Sirisinha K (2006) Chemistry of water, wastewater and analysis. Mahidol University, Bangkok

    Google Scholar 

  27. Zhang K, Kurano N, Miyachi S (2002) Optimized aeration by carbon dioxide gas for microalgal production and mass transfer characterization in a vertical flat-plate photobioreactor. Bioprocess Biosyst Eng 25(2):97–101. https://doi.org/10.1007/s00449-002-0284-y

    Article  Google Scholar 

  28. Lu S, Eiteman MA, Altman E (2009) Effect of CO2 on succinate production in dual-phase Escherichia coli fermentations. J Biotechnol 143(3):213–223. https://doi.org/10.1016/j.jbiotec.2009.07.012

    Article  Google Scholar 

  29. Cheng KK, Wu J, Wang GY, Li WY, Feng J, Zhang JA (2013) Effects of pH and dissolved CO2 level on simultaneous production of 2,3-butanediol and succinic acid using Klebsiella pneumoniae. Bioresour Technol 135:500–503. https://doi.org/10.1016/j.biortech.2012.08.100

    Article  Google Scholar 

  30. Liu Y-P, Zheng P, Sun Z-H, Ni Y, Dong J-J, Wei P (2008) Strategies of pH control and glucose-fed batch fermentation for production of succinic acid by Actinobacillus succinogenes CGMCC1593. J Chem Technol 83(5):722–729. https://doi.org/10.1002/jctb.1862

    Article  Google Scholar 

  31. Samuelov NS, Lamed R, Lowe S, Zeikus JG (1991) Influence of CO2- HCO3- levels and pH on growth, succinate production, and enzyme activities of Anaerobiospirillum succiniciproducens. Appl Environ Microbiol 57:3013–3019. https://doi.org/10.1128/AEM.57.10.3013-3019.1991

    Article  Google Scholar 

  32. Carvalho M, Roca C, Reis MA (2016) Improving succinic acid production by Actinobacillus succinogenes from raw industrial carob pods. Bioresour Technol 218:491–497. https://doi.org/10.1016/j.biortech.2016.06.140

    Article  Google Scholar 

  33. Li C, Gao S, Wang H, Li X, Ong KL, Yang X, Lin CSK (2018) Succinic acid production using a glycerol-based medium by an engineered strain of Yarrowia lipolytica: statistical optimization and preliminary economic feasibility study. Biochem Eng J 137:305–313. https://doi.org/10.1016/j.bej.2018.06.012

    Article  Google Scholar 

  34. Ong KL, Li C, Li X, Zhang Y, Xu J, Lin CSK (2019) Co-fermentation of glucose and xylose from sugarcane bagasse into succinic acid by Yarrowia lipolytica. Biochem Eng J 148:108–115. https://doi.org/10.1016/j.bej.2019.05.004

    Article  Google Scholar 

  35. Vlysidis A, Binns M, Webb C, Theodoropoulos C (2009) Utilisation of glycerol to platform chemicals within the biorefinery concept: a case for succinate production. Chem Eng Trans 58-59:1–11. https://doi.org/10.3303/CET0918087

    Article  Google Scholar 

  36. Chiang Y-S, Kuo Y-Y, Li S-Y (2018) A novel method for preparing high purity Actinobacillus succinogenes stock and its long-term acid production in a packed bed reactor. Bioresour Technol Rep 2:62–68. https://doi.org/10.1016/j.biteb.2018.04.004

    Article  Google Scholar 

  37. Alexandri M, Papapostolou H, Stragier L, Verstraete W, Papanikolaou S, Koutinas AA (2017) Succinic acid production by immobilized cultures using spent sulphite liquor as fermentation medium. Bioresour Technol 238:214–222. https://doi.org/10.1016/j.biortech.2017.03.132

    Article  Google Scholar 

  38. Luthfi AAI, Tan JP, Isa N, Bukhari NA, Shah SSM, Mahmod SS et al (2020) Multiple crystallization as a potential strategy for efficient recovery of succinic acid following fermentation with immobilized cells. Bioprocess Biosyst Eng 43:1153–1169. https://doi.org/10.1007/s00449-020-02311-x

    Article  Google Scholar 

  39. Nemati M, Webb C (2011) Immobilized cells bioreactors. In: Moo-Young M (ed) Comprehensive Biotechnology, 2nd edn. Elsevier, Amsterdam, pp 331–346

    Chapter  Google Scholar 

Download references

Acknowledgments

The authors sincerely thank the National Center for Genetic Engineering and Biotechnology (BIOTEC) for providing laboratory support.

Funding

Financial support for this research was granted by the Research and Researchers for Industries, the Thailand Research Fund, and Patum Vegetable Oil Co., Ltd. (Grant no. MSD60I0091). The study was partially supported for publication by the Faculty of Public Health, Mahidol University, Bangkok, Thailand.

Author information

Authors and Affiliations

  1. Faculty of Public Health, Department of Environmental Health Sciences, Mahidol University, 420/1 Ratchawithi RD., Ratchathewi District, Bangkok, 10400, Thailand

    Apirak Bumyut, Chatchawal Singhakant & Suwimon Kanchanasuta

  2. Biorefinery and Bioproduct Technology Research Group, National Center for Genetic Engineering and Biotechnology, 113 Thailand Science Park, Paholyothin RD., Klong Luang District, Pathumthani, 12120, Thailand

    Verawat Champreda

  3. Center of Excellence on Environmental Health and Toxicology, Bangkok, Thailand

    Chatchawal Singhakant & Suwimon Kanchanasuta

Authors
  1. Apirak Bumyut
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Verawat Champreda
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chatchawal Singhakant
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Suwimon Kanchanasuta
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Apirak conducted the experiments, analyzed and discussed the results, and wrote the manuscript.

Verawat revised and proved the manuscript for important intellectual content.

Chatchawal contributed to the final manuscript.

Suwimon supervised the project, designed the study, and finally approved the version to be submitted.

Corresponding author

Correspondence to Suwimon Kanchanasuta.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bumyut, A., Champreda, V., Singhakant, C. et al. Effects of immobilization of Actinobacillus succinogenes on efficiency of bio-succinic acid production from glycerol. Biomass Conv. Bioref. 12, 643–654 (2022). https://doi.org/10.1007/s13399-020-01069-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13399-020-01069-2

Keywords

Search

Navigation