Abstract
Ethanol and β-galactosidase production from cheese whey may significantly contribute to minimise environmental problems while producing value from low-cost raw materials. In this work, the recombinant Saccharomyces cerevisiae NCYC869-A3/pVK1.1 flocculent strain expressing the lacA gene (coding for β-galactosidase) of Aspergillus niger under ADHI promoter and terminator was used. This strain shows high ethanol and β-galactosidase productivities when grown on lactose. Batch cultures were performed using SSlactose medium with 50 g L−1 lactose in a 2-L bioreactor under aerobic and microaerophilic conditions. Temperature was maintained at 30 °C and pH 4.0. In order to determine the effect of an electric field in the fermentation profile, titanium electrodes were placed inside the bioreactor and different electric field values (from 0.5 to 2 V cm−1) were applied. For all experiments, β-galactosidase activity, biomass, protein, lactose, glucose, galactose and ethanol concentrations were measured. Finally, lag phase duration and specific growth rate were calculated. Significant changes in lag phase duration and biomass yield were found when using 2 V cm−1. Results show that the electric field enhances the early stages of fermentation kinetics, thus indicating that its application may improve industrial fermentations’ productivity. The increase in electric field intensity led to plasmid instability thus decreasing β-galactosidase production.
Similar content being viewed by others
References
Araújo, O. Q. F., Coelho, M. A. Z., Margarit, I. C. P., Vaz, C. A., & Rocha-Leão, M. H. M. (2004). Electrical stimulation of Saccharomyces cerevisiae cultures. Brazilian Journal of Microbiology, 35(1–2), 97–103.
Bailey, M. J., & Linko, M. (1990). Production of beta-galactosidase by Aspergillus oryzae in submerged bioreactor cultivation. Journal of Biotechnology, 16(1–2), 57–66.
Bartlett, P. N., Pletcher, D., & Zeng, J. (1997). Approaches to the integration of electrochemistry and biotechnology. Journal of the Electrochemical Society, 144(11), 3705–3710.
Baysal, A. H., & Icier, F. (2010). Inactivation kinetics of Alicyclobacillus acidoterrestris spores in orange juice by ohmic heating: Effects of voltage gradient and temperature on inactivation. Journal of Food Protection, 73(2), 299–304.
Castro, I., Macedo, B., Teixeira, J. A., & Vicente, A. A. (2004). The effect of electric field on important food-processing enzymes: Comparison of inactivation kinetics under conventional and ohmic heating. Journal of Food Science, 69(9), C696–C701.
Cho, H. Y., Yousef, A. E., & Sastry, S. K. (1996). Growth kinetics of Lactobacillus acidophilus under ohmic heating. Biotechnology and Bioengineering, 49(3), 334–340.
Domingues, L., Lima, N., & Teixeira, J. A. (2005). Aspergillus niger beta-galactosidase production by yeast in a continuous high cell density reactor. Process Biochemistry, 40(3–4), 1151–1154.
Domingues, L., Oliveira, C., Castro, I., Lima, N., & Teixeira, J. A. (2004). Production of beta-galactosidase from recombinant Saccharomyces cerevisiae grown on lactose. Journal of Chemical Technology and Biotechnology, 79(8), 809–815.
Domingues, L., Onnela, M. L., Teixeira, J. A., Lima, N., & Penttilä, M. (2000). Construction of a flocculent brewer's yeast strain secreting Aspergillus niger beta-galactosidase. Applied Microbiology and Biotechnology, 54(1), 97–103.
Domingues, L., Teixeira, J. A., Penttilä, M., & Lima, N. (2002). Construction of a flocculent Saccharomyces cerevisiae strain secreting high levels of Aspergillus niger beta-galactosidase. Applied Microbiology and Biotechnology, 58(5), 645–650.
Futcher, A. B., & Cox, B. S. (1984). Copy number and the stability of 2-micron circle-based artificial plasmids of Saccharomyces cerevisiae. Journal of Bacteriology, 157(1), 283–290.
Icier, F., & Bozkurt, H. (2011). Ohmic heating of liquid whole egg: Rheological behaviour and fluid dynamics. Food and Bioprocess Technology, in press. doi:10.1007/s11947-009-0229-4.
Kumar, V., Ramakrishnan, S., Teeri, T. T., Knowles, J. K., & Hartley, B. S. (1992). Saccharomyces cerevisiae cells secreting an Aspergillus niger β-galactosidase grow on whey permeate. Biotechnology, 10, 82–85.
Loghavi, L., Sastry, S. K., & Yousef, A. E. (2007). Effect of moderate electric field on the metabolic activity and growth kinetics of Lactobacillus acidophilus. Biotechnology and Bioengineering, 98(4), 872–881.
Loghavi, L., Sastry, S. K., & Yousef, A. E. (2008). Effect of moderate electric field frequency on growth kinetics and metabolic activity of Lactobacillus acidophilus. Biotechnology Progress, 24(1), 148–153.
Loghavi, L., Sastry, S. K., & Yousef, A. E. (2009). Effect of moderate electric field frequency and growth stage on the cell membrane permeability of Lactobacillus acidophilus. Biotechnology Progress, 25(1), 85–94.
Ludwig, D. L., & Bruschi, C. V. (1991). The 2-micron plasmid as a nonselectable, stable, high copy number yeast vector. Plasmid, 25(2), 81–95.
Machado, L. F., Pereira, R. N., Martins, R. C., Teixeira, J. A., & Vicente, A. A. (2010). Moderate electric fields can inactivate Escherichia coli at room temperature. Journal of Food Engineering, 96(4), 520–527.
Miller, G. L. (1959). Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 31(3), 426–428.
Oliveira, C., Guimarães, P. M., & Domingues, L. (2011). Recombinant microbial systems for improved β-galactosidase production and biotechnological applications. Biotechnology Advances, in press. doi:10.1016/j.biotechadv.2011.03.008.
Oliveira, C., Teixeira, J. A., Lima, N., Da Silva, N. A., & Domingues, L. (2007). Development of stable flocculent Saccharomyces cerevisiae strain for continuous Aspergillus niger beta-galactosidase production. Journal of Bioscience and Bioengineering, 103(4), 318–324.
Panesar, P. S., Panesar, R., Singh, R. S., Kennedy, J. F., & Kumar, H. (2006). Microbial production, immobilization and applications of beta-D-galactosidase. Journal of Chemical Technology and Biotechnology, 81(4), 530–543.
Pereira, R., Martins, J., Mateus, C., Teixeira, J. A., & Vicente, A. A. (2007). Death kinetics of Escherichia coli in goat milk and Bacillus licheniformis in cloudberry jam treated by ohmic heating. Chemical Papers, 61(2), 121–126.
Pereira, R. N., & Vicente, A. A. (2010). Environmental impact of novel thermal and non-thermal technologies in food processing. Food Research International, 43, 1936–1943.
Rowley, B. A. (1972). Electrical current effects on Escherichia coli growth rates. Proceedings of the Society for Experimental Biology and Medicine, 139(3), 929–934.
Stacey, M., Stickley, J., Fox, P., Statler, V., Schoenbach, K., Beebe, S. J., et al. (2003). Differential effects in cells exposed to ultra-short, high intensity electric fields: Cell survival, DNA damage, and cell cycle analysis. Mutation Research, Genetic Toxicology and Environmental Mutagenesis, 542(1–2), 65–75.
Sun, H. X., Kawamura, S., Himoto, J. I., Itoh, K., Wada, T., & Kimura, T. (2008). Effects of ohmic heating on microbial counts and denaturation of proteins in milk. Food Science and Technology Research, 14(2), 117–123.
Wei, P., Li, Z., Lin, Y., He, P., & Jiang, N. (2007). Improvement of the multiple-stress tolerance of an ethanologenic Saccharomyces cerevisiae strain by freeze–thaw treatment. Biotechnology Letters, 29(10), 1501–1508.
Yildiz, H., & Baysal, T. (2006). Effects of alternative current heating treatment on Aspergillus niger, pectin methylesterase and pectin content in tomato. Journal of Food Engineering, 75(3), 327–332.
Zell, M., Lyng, J. G., Morgan, D. J., & Cronin, D. A. (2011). Quality evaluation of an ohmically cooked ham product. Food and Bioprocess Technology, in press. doi:10.1007/s11947-009-0281-0.
Acknowledgements
The authors gratefully acknowledge Fundação para a Ciência e a Tecnologia (Portugal) for the scholarships SFRH/BD/11230/2002 and SFRH/BDP/63831/2009 granted to authors I. Castro and C. Oliveira, respectively.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Castro, I., Oliveira, C., Domingues, L. et al. The Effect of the Electric Field on Lag Phase, β-Galactosidase Production and Plasmid Stability of a Recombinant Saccharomyces cerevisiae Strain Growing on Lactose. Food Bioprocess Technol 5, 3014–3020 (2012). https://doi.org/10.1007/s11947-011-0609-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11947-011-0609-4