Production and biochemical characterization of the recombinant Boophilus microplus Bm95 antigen from Pichia pastoris

Boue, O., Farnos, O., Gonzalez, A., Fernandez, R., Acosta, J. A., Valdes, R., Gonzalez, L. J., Guanche, Y., Izquierdo, G., Suarez, M., Dominguez, I., Machado, H., Rodriguez, M. and Lleonart, R. (2004) Production and biochemical characterization of the recombinant Boophilus microplus Bm95 antigen from Pichia pastoris. Experimental and Applied Acarology, 32 1-2: 119-128. doi:10.1023/B:APPA.0000018199.87122.e4


Author Boue, O.
Farnos, O.
Gonzalez, A.
Fernandez, R.
Acosta, J. A.
Valdes, R.
Gonzalez, L. J.
Guanche, Y.
Izquierdo, G.
Suarez, M.
Dominguez, I.
Machado, H.
Rodriguez, M.
Lleonart, R.
Title Production and biochemical characterization of the recombinant Boophilus microplus Bm95 antigen from Pichia pastoris
Journal name Experimental and Applied Acarology   Check publisher's open access policy
ISSN 0168-8162
Publication date 2004
Sub-type Article (original research)
DOI 10.1023/B:APPA.0000018199.87122.e4
Volume 32
Issue 1-2
Start page 119
End page 128
Total pages 10
Language eng
Abstract The new antigen Bm95 from the cattle tick Boophilus microplus was recently isolated, cloned and expressed in the methylotrophic yeast Pichia pastoris. The recombinant protein has shown to induce protection in cattle against infestations of B. microplus under controlled and production conditions. In this paper we report the production and large-scale purification of the Bm95 protein, following a simple and cost-effective process. The antigen was obtained highly aggregated, forming particles ranging from 26 to 30 nm and with purity higher than 80%. The process yield was 0.55 g of pure Bm95 protein per liter of culture. The 98% of the primary structure of the recombinant protein was verified by mass spectrometry. Three amino acid changes in comparison with the sequence deduced from cDNA were detected by LC-MS/MS. The antigen was also obtained N-glycosylated, as previously reported for heterologous protein expression in P. pastoris.
Keyword Bm86
Bm95
Boophilus microplus
Cattle tick
Pichia pastoris
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Queensland Alliance for Agriculture and Food Innovation
 
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