Abstract
Baker’s yeast was disrupted in a 1.4-L stainless steel horizontal bead mill under a continuous recycle mode using 0.3 mm diameter zirconia beads as abrasive. A single pass in continuous mode bead mill operation liberates half of the maximally released protein. The maximum total protein release can only be achieved after passaging the cells 5 times through the disruption chamber. The degree of cell disruption was increased with the increase in feeding rate, but the total protein release was highest at the middle range of feeding rate (45 L/h). The total protein release was increased with an increase in biomass concentration from 10 to 50% (w/v). However, higher heat dissipation as a result of high viscosity of concentrated biomass led to the denaturation of labile protein such as glucose 6-phosphate dehydrogenase (G6PDH). As a result the highest specific activity of G6PDH was achieved at biomass concentration of 20% (ww/v). Generally, the degree of cell disruption and total protein released were increased with an increase in impeller tip speed, but the specific activity of G6PDH was decreased substantially at higher impeller tip speed (14 m/s). Both the degree of cell disruption and total protein release increased, as the bead loading increased from 75 to 85% (v/v). Hence, in order to obtain a higher yield of labile protein such as G6PDH, the yeast cell should not be disrupted at biomass concentration and impeller tip speed higher than 20% (w/v) and 10 m/s, respectively.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Middlelberg, A. P. J. (1995) Process scale disruption of microorganisms.Biotechnol. Adv. 13: 491–550.
Limon-Lason, J., J. Hoare, C. B. Orsborn, D. J. Doyle, and P. Dunnill (1979) Reactor properties of a high-speed bead mill for microbial cell rupture.Biotechnol. Bioeng. 21: 745–774.
Schutte, H., K. H. Kroner, H. Hustedt, and M.-R. Kula (1983) Experiences with a 20 litre industrial bead mill for the disruption of microorganisms.Enzyme Microb. Technol. 5: 143–148.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analyt. Biochem. 72: 248–254.
Bergmeyer, H. U., M. Grabl, and H. E. Walter (1983) Biochemical reagents for general use: Enzymes. pp. 222–223. In: Bergmeyer, U. (ed.).Methods in Enzymatic Analysis. Academic Press, NY, USA.
Kula, M.-R. and H. Schütte. (1987) Purification of proteins and the disruption of microbial cells.Biotechnol. Prog. 3: 31–42.
Ricci-Silva, M. E., M. Vitolo, and J. Abrahão-Neto (2000) Protein and glucose 6-phosphate dehydrogenase releasing from baker’s yeast cells disrupted by a vertical bead mill.Process Biochemistry 35: 831–835.
Garrido, F., U. C. Banerjee, Y. Chisti, and M. Moo-Young (1994) Disruption of a recombinant yeast for the release of β-galactosidase.Bioseparation 4: 319–328.
Garcia, F. A. P. (1993) Cell wall disruption. pp.47–67. In: Kennedy, J. F. and J. M. S. Cabral (eds.).Recovery Processes for Biological Materials. John Wiley and Sons. Chichester, UK.
Melendres, A. V., H. Honda, N. Shiragami, and H. Unno (1991) A kinetic analysis of cell disruption by bead mill.Bioseparation 2: 231–236.
Melendres, A. V., H. Unno, N. Shiragami, and H. Honda (1992) A concept of critical velocity for cell disruption by bead mill.J. Chemical Eng. Japan 25: 354–356.
Mario Canales, J. A., L. H. Buxado, and E. Antonio (1998) Mechanical disruption ofPichia pastoris yeast to recover the recombinant glycoprotein Bm86. In:Genetic Engineering and Biotechnol. Havana, Cuba.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Mei, C.Y., Ti, T.B., Ibrahim, M.N. et al. The disruption ofSaccharomyces cerevisiae cells and release of glucose 6-phosphate dehydrogenase (G6PDH) in a horizontal dyno bead mill operated in continuous recycling mode. Biotechnol. Bioprocess Eng. 10, 284–288 (2005). https://doi.org/10.1007/BF02932027
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02932027