Secretion, purification and activity of two recombinant pepper endo-β-1,4-glucanases expressed in the yeast Pichia pastoris

被引：23

作者：

Ferrarese, L ^{[1
]}

Trainotti, L ^{[1
]}

Gattolin, S ^{[1
]}

Casadoro, G ^{[1
]}

机构：

[1] Univ Padua, Dept Biol, I-35121 Padua, Italy

来源：

FEBS LETTERS | 1998年 / 422卷 / 01期

关键词：

endo-beta-1,4-glucanase; Pichia pastoris; heterologous expression; Capsicum annuum;

D O I：

10.1016/S0014-5793(97)01592-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Two pepper endo-beta-1,4-glucanases, involved in fruit softening (cCel1) and leaf and flower abscission (cCel2), have been expressed in Pichia pastor is. Secretion was obtained by using either the mouse a-factor signal(cCel1) or the native signal sequence (cCel2), Times for optimal expression of the two proteins were different and cCel2 appeared very sensitive to proteolytic degradation, A one-step purification protocol yielded cCel2 in a pure form, while an additional chromatography step was necessary to purify cCel1, The two recombinant proteins are highly active and able to degrade carboxymethylcellulose in viscometric assays. Moreover, they have both a molecular mass (54 kDa) and an isoelectric point (7.2 for cCel2, and 8.5 for cCel1) equal to those of the native proteins, thus suggesting that post-translational modifications have properly occurred. (C) 1998 Federation of European Biochemical Societies.

引用

页码：23 / 26

页数：4

共 21 条

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