Substrate Type and CO Addition Significantly Influence Succinic Acid Production of Basfia Succiniciproducens

Overview

Journal Biotechnol Lett

Specialties Biochemistry
Biotechnology

Date 2023 Jul 3

PMID 37395870

Authors

Marta Balazs

Hunor Bartos

Szabolcs Lanyi

Zsolt Bodor

Ildiko Miklossy

Affiliations

Soon will be listed here.

Abstract

Metabolic engineering has shown that optimizing metabolic pathways' fluxes for industrial purposes requires a methodical approach. Accordingly, in this study, in silico metabolic modeling was utilized to characterize the lesser-known strain Basfia succiniciproducens under different environmental conditions, followed by the use of industrially relevant substrates for succinic acid synthesis. Based on RT-qPCR carried out in flask experiments, we discovered a relatively large difference in the expression levels of ldhA gene compared to glucose in both xylose and glycerol cultures. In bioreactor-scale fermentations, the impact of different gas phases (CO, CO/AIR) on biomass yield, substrate consumption, and metabolite profiles was also investigated. In the case of glycerol, the addition of CO increased biomass as well as target product formation, while using CO/AIR gas phase resulted in higher target product yield (0.184 mM⋅mM). In case of xylose, using CO alone would result in higher succinic acid production (0.277 mM⋅mM). The promising rumen bacteria, B. succiniciproducens, has shown to be suitable for succinic acid production from both xylose and glycerol. As a result, our findings present new opportunities for broadening the range of raw materials used in this significant biochemical process. Our study also sheds light on fermentation parameter optimization for this strain, namely that, CO/AIR supply has a positive effect on target product formation.

References

Murarka A, Dharmadi Y, Yazdani S, Gonzalez R . Fermentative utilization of glycerol by Escherichia coli and its implications for the production of fuels and chemicals. Appl Environ Microbiol. 2007; 74(4):1124-35. PMC: 2258577. DOI: 10.1128/AEM.02192-07. View

Ma J, Jiang M, Chen K, Xu B, Liu S, Wei P . Succinic acid production with metabolically engineered E. coli recovered from two-stage fermentation. Biotechnol Lett. 2010; 32(10):1413-8. DOI: 10.1007/s10529-010-0313-x. View

Stylianou E, Pateraki C, Ladakis D, Cruz-Fernandez M, Latorre-Sanchez M, Coll C . Evaluation of organic fractions of municipal solid waste as renewable feedstock for succinic acid production. Biotechnol Biofuels. 2020; 13:72. PMC: 7160979. DOI: 10.1186/s13068-020-01708-w. View

Kim H, Jeong H, Lee S . Short-Term Adaptation Modulates Anaerobic Metabolic Flux to Succinate by Activating ExuT, a Novel D-Glucose Transporter in . Front Microbiol. 2020; 11:27. PMC: 6989600. DOI: 10.3389/fmicb.2020.00027. View

Scholten E, Renz T, Thomas J . Continuous cultivation approach for fermentative succinic acid production from crude glycerol by Basfia succiniciproducens DD1. Biotechnol Lett. 2009; 31(12):1947-51. DOI: 10.1007/s10529-009-0104-4. View

Becker S, Feist A, Mo M, Hannum G, Palsson B, Herrgard M . Quantitative prediction of cellular metabolism with constraint-based models: the COBRA Toolbox. Nat Protoc. 2007; 2(3):727-38. DOI: 10.1038/nprot.2007.99. View

Kuhnert P, Scholten E, Haefner S, Mayor D, Frey J . Basfia succiniciproducens gen. nov., sp. nov., a new member of the family Pasteurellaceae isolated from bovine rumen. Int J Syst Evol Microbiol. 2009; 60(Pt 1):44-50. DOI: 10.1099/ijs.0.011809-0. View

Song H, Lee J, Choi S, You J, Hong W, Lee S . Effects of dissolved CO2 levels on the growth of Mannheimia succiniciproducens and succinic acid production. Biotechnol Bioeng. 2007; 98(6):1296-304. DOI: 10.1002/bit.21530. View

Pennacchio A, Ventorino V, Cimini D, Pepe O, Schiraldi C, Inverso M . Isolation of new cellulase and xylanase producing strains and application to lignocellulosic biomasses hydrolysis and succinic acid production. Bioresour Technol. 2018; 259:325-333. DOI: 10.1016/j.biortech.2018.03.027. View

10.

Lee S, Song H, Lee S . Genome-based metabolic engineering of Mannheimia succiniciproducens for succinic acid production. Appl Environ Microbiol. 2006; 72(3):1939-48. PMC: 1393240. DOI: 10.1128/AEM.72.3.1939-1948.2006. View

11.

Yin X, Li J, Shin H, Du G, Liu L, Chen J . Metabolic engineering in the biotechnological production of organic acids in the tricarboxylic acid cycle of microorganisms: Advances and prospects. Biotechnol Adv. 2015; 33(6 Pt 1):830-41. DOI: 10.1016/j.biotechadv.2015.04.006. View

12.

Lee P, Lee W, Lee S, Chang H . Succinic acid production with reduced by-product formation in the fermentation of Anaerobiospirillum succiniciproducens using glycerol as a carbon source. Biotechnol Bioeng. 2000; 72(1):41-8. View

13.

Cimini D, Argenzio O, Dambrosio S, Lama L, Finore I, Finamore R . Production of succinic acid from Basfia succiniciproducens up to the pilot scale from Arundo donax hydrolysate. Bioresour Technol. 2016; 222:355-360. DOI: 10.1016/j.biortech.2016.10.004. View

14.

Ahn J, Jang Y, Lee S . Production of succinic acid by metabolically engineered microorganisms. Curr Opin Biotechnol. 2016; 42:54-66. DOI: 10.1016/j.copbio.2016.02.034. View

15.

Lee P, Lee S, Chang H . Kinetic study on succinic acid and acetic acid formation during continuous cultures of Anaerobiospirillum succiniciproducens grown on glycerol. Bioprocess Biosyst Eng. 2009; 33(4):465-71. DOI: 10.1007/s00449-009-0355-4. View

16.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

17.

Becker J, Reinefeld J, Stellmacher R, Schafer R, Lange A, Meyer H . Systems-wide analysis and engineering of metabolic pathway fluxes in bio-succinate producing Basfia succiniciproducens. Biotechnol Bioeng. 2013; 110(11):3013-23. DOI: 10.1002/bit.24963. View

18.

Barcelos M, Ramos C, Kuddus M, Rodriguez-Couto S, Srivastava N, Ramteke P . Enzymatic potential for the valorization of agro-industrial by-products. Biotechnol Lett. 2020; 42(10):1799-1827. DOI: 10.1007/s10529-020-02957-3. View

19.

Scholten E, Dagele D . Succinic acid production by a newly isolated bacterium. Biotechnol Lett. 2008; 30(12):2143-6. DOI: 10.1007/s10529-008-9806-2. View

20.

Lee P, Lee S, Hong S, Chang H, Park S . Biological conversion of wood hydrolysate to succinic acid by Anaerobiospirillum succiniciproducens. Biotechnol Lett. 2003; 25(2):111-4. DOI: 10.1023/a:1021907116361. View