Abstract
The size distribution and density of two types of protein inclusion bodies arising from the synthesis in E. coli of recombinant calf prochymosin and γ–interferon were examined. A combination of electrical sensing zone and centrifugal sedimentation techniques were used to characterize the particles. The mean size and standard deviation (sd) of the distributions for the γ–interferon and prochymosin–based inclusion bodies were 0.81 μm, sd 0.17 μm, and 1.28 μm, sd 0.46 μm, respectively. Sedimentation studies indicated that the density of the protein inclusion bodies increases with the density of the suspending solvent. These density variations are consistent with the bodies having a voidage, accessible to the suspending fluid of 70% (γ–interferon) and 85% (prochymosin) of the total volume.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Marston, F.A.O., Lowe, P.A., Doel, M.T., Schoemaker, J.M., White, S., and Angal, S. 1984. Purification of calf prochymosin (prorennin) synthesized in Escherichia coli. Bio/Technology 2:800–804.
Schoemaker, J.M., Brasnett, A.H., and Marston, F.A.O. 1985. Examination of calf prochymosin accumulation in Escherichia coli: disulphide linkages are a structural component of prochymosin-containing inclusion bodies. EMBO Journal 4:775–780.
Schoner, R.G., Ellis, L.F., and Schoner, B.E. 1985. Isolation and purification of protein granules from Escherichia coli cells over-producing bovine growth hormone. Bio/Technology 3:151–154.
Lee, S.G., Eckblad, W. and Bulla, Jr, L.A. 1985. Diversity of protein inclusion bodies and identification of mosquitocidal protein in Bacillus thuringiensis subsp. israelensis. Biochem. Biophys. Res. Comm. 126:953–960.
Oppenheimer, L.E. 1983. Interpretation of disc centrifuge data. J. Coll. Int. Sci. 92:350–357.
Heiss, J.F. and Coull, J. 1952. The effect of orientation and shape on the settling velocity of non-isometric particles in a viscous medium. Chem. Eng. Prog. 48:133–140.
Bell, D.J., Heywood-Waddington, D., Hoare, M., and Dunnill, P. 1982. The density of protein precipitates and its effect on centrifugal sedimentation. Biotech. Bioeng. 24:127–141.
Matthews, B.W. 1974. Determination of molecular weight from protein crystals. J. Mol. Biol. 82:513–526.
Matthews, B.W. 1968. Solvent content of protein crystals. J. Mol. Biol. 33:491–497.
Langvanker, A.L. and Gemmel, R.S. 1968. A size-density relationship for flocs. J. Am. Water Works Assoc. 9:1040–1046.
Emtage, J.S., Angal, S., Doel, M.T., Harris, T.J.R., Jenkins, B., Lilley, G., and Lowe, P. 1983. Synthesis of calf prochymosin (prorennin) in Escherichia coli. Proc. Natl. Acad. Sci. USA, 80:3671–3675.
Karuhn, R.F. and Berg, R.H. 1982. Practical Aspects of Electrozone Size Analysis, Particle Data Laboratories Ltd. Elmhurst, Illinois, USA.
Sheeler, P. 1981. Centrifugation in Biology and Medical Science. J. Wiley and Sons, NY.
Rickwood, D. 1982. Nycodenz. Nyegaard and Co. Oslo, Norway.
Joyce Loebl Manual. Joyce Loebl Ltd., Gateshead, England.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Taylor, G., Hoare, M., Gray, D. et al. Size and Density of Protein Inclusion Bodies. Nat Biotechnol 4, 553–557 (1986). https://doi.org/10.1038/nbt0686-553
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt0686-553
This article is cited by
-
Predictive approaches to guide the expression of recombinant vaccine targets in Escherichia coli: a case study presentation utilising Absynth Biologics Ltd. proprietary Clostridium difficile vaccine antigens
Applied Microbiology and Biotechnology (2021)
-
Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process
Microbial Cell Factories (2015)
-
Current state and recent advances in biopharmaceutical production in Escherichia coli, yeasts and mammalian cells
Journal of Industrial Microbiology and Biotechnology (2013)
-
Isolation of cell-free bacterial inclusion bodies
Microbial Cell Factories (2010)
-
Friendly production of bacterial inclusion bodies
Korean Journal of Chemical Engineering (2010)