Overexpression of juvenile hormone binding protein in bacteria and Pichia pastoris

被引：7

作者：

Grzelak, K

Kludkiewicz, B

Kolomiets, LI

Debski, J

Dadlez, M

Lalik, A

Ozyhar, A

Kochman, M ^{[1
]}

机构：

[1] Wroclaw Univ Technol, Inst Organ Chem Biochem & Biotechnol, Div Biochem, PL-50370 Wroclaw, Poland

[2] Polish Acad Sci, Inst Biochem & Biophys, PL-02532 Warsaw, Poland

[3] Natl Acad Sci Ukraine, Inst Mol Biol & Genet, Kiev, Ukraine

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2003年 / 31卷 / 02期

关键词：

JHBP; Pichia pastoris; recombinant JHBP; Galleria mellonella;

D O I：

10.1016/S1046-5928(03)00192-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Galleria mellonella juvenile hormone binding protein (JHBP) is a single chain glycoprotein with two disulfide bonds and a molecular mass of 25,880 Da. This report describes the expression of JHBP in bacteria and yeast cells (Pichia pastoris). The expression in bacteria was low and the protein was rapidly degraded upon cell lysis. The expression of His(8)-tagged rJHBP (His(8)-rJHBP) in P. pastoris was high and the non-degraded protein was purified to homogeneity with high yield in a one-step immobilized Ni++ affinity chromatography. His(8)-rJHBP from P. pastoris contains one JH III binding site with K-D of 3.7 +/- 1.3 x 10(-7) M. The results suggest that P. pastoris is the preferred system for expression of His8-rJHBP in non-degraded fully active form. (C) 2003 Elsevier Inc. All rights reserved.

引用

页码：173 / 180

页数：8

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