Structural Characterization of Potato Protease Inhibitor I (Cv. Bintje) after Expression in Pichia pastoris
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- 作者：Lambertus A. M. van den Broek ; Laurice Pouvreau ; Gijs Lommerse ; Bert Schipper ; Gerrit A. van Koningsveld ; and Harry Gruppen
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2004
- 出版时间：July 28, 2004
- 年：2004
- 卷：52
- 期：15
- 页码：4928 - 4934
- 全文大小：139K
- 年卷期：v.52,no.15(July 28, 2004)
- ISSN：1520-5118
文摘
In the present study the structural properties of potato protease inhibitor 1 (PI-1) were studied as afunction of temperature to elucidate its precipitation mechanism upon heating. A cDNA coding forPI-1 from cv. Bintje was cloned and expressed in Pichia pastoris. Using the recombinant PI-1 it wassuggested that PI-1 behaves as a hexameric protein rather than as a pentamer, as previously proposedin the literature. The recombinant protein seems either to have a predominantly unordered structureor to belong to the 
-II proteins. Differential scanning calorimetry analysis of PI-1 revealed that itsthermal unfolding occurs via one endothermic transition in which the hexameric PI-1 probably unfolds,having a dimer instead of a monomer as cooperative unit. The transition temperature for therecombinant PI-1 was 88 
C. Similar results were obtained for a partially purified pool of native PI-1from cv. Bintje.Keywords: Potato; Solanum tuberosum; protease inhibitor; Pichia pastoris; recombinant PI-1
-II proteins. Differential scanning calorimetry analysis of PI-1 revealed that itsthermal unfolding occurs via one endothermic transition in which the hexameric PI-1 probably unfolds,having a dimer instead of a monomer as cooperative unit. The transition temperature for therecombinant PI-1 was 88 C. Similar results were obtained for a partially purified pool of native PI-1from cv. Bintje.Keywords: Potato; Solanum tuberosum; protease inhibitor; Pichia pastoris; recombinant PI-1