Thermal stabilization of penicillolysin, a thermolabile 19 kDa Zn2+-protease, obtained by site-directed mutagenesis

被引：7

作者：

Doi, Y

Akiyama, H

Yamada, Y

Ee, CE

Lee, BR

Ikeguchi, M

Ichishima, E ^{[1
]}

机构：

[1] Soka Univ, Grad Sch Bioengn, Lab Mol Enzymol, Hachioji, Tokyo 1928577, Japan

[2] Tohoku Univ, Grad Sch Agr Sci, Lab Mol Enzymol, Aoba Ku, Sendai, Miyagi 9818555, Japan

来源：

PROTEIN ENGINEERING DESIGN & SELECTION | 2004年 / 17卷 / 03期

关键词：

aspzincin; M35; protease; mutagenesis; penicillolysin; thermal stabilization; Zn2+-protease;

D O I：

10.1093/protein/gzh034

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：

Penicillolysin is a member of the clan NIX and the family of M35 proteases. The enzyme is a thermolabile Zn2+-protease from Penicillium citrinum with a unique substrate profile. We expressed recombinant penicillolysin in Aspergillus oryzae and generated several site-directed mutants, R33E/E60R, A167E and T81P, with the intention of exploring thermal stabilization of this protein. We based our choice of mutations on the structures of homologous thermally stable enzymes, deuterolysin (EC 3.4.24.39) from A.oryzae and a peptidyl-Lys metallopeptidase (GfMEP) from the edible mushroom Grifora frondsa. The resulting mutant proteins exhibited comparable catalytic efficiency to the wild-type enzyme and some showed a higher tolerance to temperature.

引用

页码：261 / 266

页数：6

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