Fumaric acid production using renewable resources from biodiesel and cane sugar production processes
The microbial production of fumaric acid by Rhizopus arrhizus NRRL 2582 has been evaluated using soybean cake from biodiesel production processes and very high polarity (VHP) sugar from sugarcane mills. Soybean cake was converted into a nutrient-rich hydrolysate via a two-stage bioprocess involving crude enzyme production via solid state fermentations (SSF) of either Aspergillus oryzae or R. arrhizus cultivated on soybean cake followed by enzymatic hydrolysis of soybean cake. The soybean cake hydrolysate produced using crude enzymes derived via SSF of R. arrhizus was supplemented with VHP sugar and evaluated using different initial free amino nitrogen (FAN) concentrations (100, 200, and 400 mg/L) in fed-batch cultures for fumaric acid production. The highest fumaric acid concentration (27.3 g/L) and yield (0.7 g/g of total consumed sugars) were achieved when the initial FAN concentration was 200 mg/L. The combination of VHP sugar with soybean cake hydrolysate derived from crude enzymes produced by SSF of A. oryzae at 200 mg/L initial FAN concentration led to the production of 40 g/L fumaric acid with a yield of 0.86 g/g of total consumed sugars. The utilization of sugarcane molasses led to low fumaric acid production by R. arrhizus, probably due to the presence of various minerals and phenolic compounds. The promising results achieved through the valorization of VHP sugar and soybean cake suggest that a focused study on molasses pretreatment could lead to enhanced fumaric acid production.
KeywordsBioprocess Fumaric acid Cane sugar Molasses Rhizopus arrhizus Soybean cake
This work was funded by Petrobras (Brazil) (project 2012/00320-2) and the National Council for Scientific and Technological Development of the Ministry of Science, Technology, and Innovation (CNPq/MCTI) through the Special Visiting Researcher fellowship (process number: 313772/2013-4).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Ashraf S, Sikander A, Haq I (2015) Acidic pre-treatment of sugarcane molasses for molasses for citric acid production by Aspergillus niger NG-4. Int J Curr Microbiol Appl Sci 4:584–595Google Scholar
- Delgenes JP, Moletta R, Navarro JM (1996) Effects of lignocellulose degradation products on ethanol fermentations of glucose and xylose by Saccharomyces cerevisiae, Zymomonas mobilis, Pichia stipitis, and Candida shehatae. Enzym Microb Technol 19:220–225. https://doi.org/10.1016/0141-0229(95)00237-5 CrossRefGoogle Scholar
- Dimou C, Kopsahelis N, Papadaki A, Papanikolaou S, Kookos IK, Mandala I, Koutinas AA (2015) Wine lees valorization: Biorefinery development including production of a generic fermentation feedstock employed for poly(3-hydroxybutyrate) synthesis. Food Res Int 73:81–87. https://doi.org/10.1016/j.foodres.2015.02.020 CrossRefGoogle Scholar
- Eurostat (2016) Renewable energy statistics. http://ec.europa.eu/eurostat/statistics-explained/index.php/Renewable_energy_statistics. Accessed 15 October 2017
- FAOSTAT (2014) http://www.fao.org/faostat/en/#data/QC. Accessed 16 October 2017
- Goldberg I, Lonberg-Holm K, Bagley EA, Stieglitz B (1983) improved conversion of fumarate to succinate Escherichia coli strains amplified for fumarate reductase. Appl Environ Microbiol 1838–1847Google Scholar
- Harland BF, Harland J (1980) Fermentative reduction of phytate in rye, white and whole wheat breads. Cereal Chem 57:226–229Google Scholar
- Ito S, Barchi AC, Escaramboni B, de Oliva Neto P, Herculano RD, Azevedo Borges F, Romeiro Miranda MC, Fernández Núñez EG (2017) UV/Vis spectroscopy combined with chemometrics for monitoring solid-state fermentation with Rhizopus microsporus var. oligosporus. J Chem Technol Biotechnol 92:2563–2572. https://doi.org/10.1002/jctb.5271 CrossRefGoogle Scholar
- Kachrimanidou V, Kopsahelis N, Chatzifragkou A, Papanikolaou S, Yanniotis S, Kookos I, Koutinas AA (2013) Utilisation of by-products from sunflower-based biodiesel production processes for the production of fermentation feedstock. Waste Biomass Valor 4:529–537. https://doi.org/10.1007/s12649-012-9191-x CrossRefGoogle Scholar
- Koutinas AA, Vlysidis A, Pleissner D, Kopsahelis N, Lopez Garcia I, Kookos IK, Papanikolaou S, Kwan TH, Lin CSK (2014) Valorization of industrial waste and by-product streams via fermentation for the production of chemicals and biopolymers. Chem Soc Rev 43:2587–2627. https://doi.org/10.1039/c3cs60293a CrossRefGoogle Scholar
- Lie S (1973) The EBC-ninhydrin method for determination of free alpha amino nitrogen. J I Brewing 79:37–41. https://doi.org/10.1002/j.2050-0416.1973.tb03495.x CrossRefGoogle Scholar
- Papadaki A, Androutsopoulos N, Patsalou M, Koutinas M, Kopsahelis N, de Castro AM, Papanikolaou S, Koutinas AA (2017) Biotechnological production of Fumaric acid: the effect of morphology of Rhizopus arrhizus NRRL 2582. Fermentation 3:33. https://doi.org/10.3390/fermentation3030033 CrossRefGoogle Scholar
- Rauf A, Irfan M, Nadeem M, Ahmed I, Iqbal HMN (2010) Optimization of Growth Conditions for Acidic Protease Production from Rhizopus oligosporus through Solid State Fermentation of Sunflower Meal. WASET International Journal of Biotechnology and Bioengineering 4(12). dai: https://doi.org/10.1999/1307-6892/12762
- Rhodes RA, Moyer AJ, Smith ML, Kelley SE (1959) Production of fumaric acid by Rhizopus arrhizus. Appl Microbiol 7:74–80Google Scholar
- Tsouko E, Kachrimanidou V, Dos Santos AF, do Nascimento Vitorino Lima ME, Papanikolaou S, de Castro AM, Freire DM, Koutinas AA (2017) Valorization of By-Products from Palm Oil Mills for the Production of Generic Fermentation Media for Microbial Oil Synthesis. Appl Biochem Biotechnol 181:1241–1256. https://doi.org/10.1007/s12010-016-2281-7 CrossRefGoogle Scholar
- Zhou Y, Du J, Tsao GT (2000) Mycellial Pellet Formation by Rhizopus oryzae ATCC 20344. Appl Biochem Biotechnol 84–86:779–89Google Scholar