Redesign the α/β fold to enhance the stability of mannanase Man23 from bacillus subtilis
作者:
Zhou, Hai-Yan;Pan, Hong-Ya;Rao, Li-Qun;Wu, Yong-Yao*
期刊:
Applied Biochemistry and Biotechnology,2011年163(1):186-194 ISSN:0273-2289
通讯作者:
Wu, Yong-Yao
作者机构:
[Zhou, Hai-Yan] Hunan Agr Univ, Ctr Anal & Measurement, Changsha 410128, Hunan, Peoples R China.;[Rao, Li-Qun; Wu, Yong-Yao] Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Hunan, Peoples R China.;[Pan, Hong-Ya] Hunan Agr Univ, Coll Foreign Language, Changsha 410128, Hunan, Peoples R China.
通讯机构:
[Wu, Yong-Yao] H;Hunan Agr Univ, Coll Biosci & Biotechnol, Changsha 410128, Hunan, Peoples R China.
关键词:
Mannanase;Protein stability;Rational design;α/β Fold
摘要:
In this work, we engineered the α/β fold of mannanase Man23 based on its molecular structure analysis to obtain more stable variants. By introducing 31 single-site mutations in the α/β fold and shuffling them, the incorporation of four mutations (K178R, K207R, N340R, and S354R) displayed a good balance between high activity and stability at higher temperature and broader pH. This quartet variant was characterized by an almost threefold increased activity and a sevenfold increased stability compared to native mannanase Man23. Our results suggest that such work is safe to increase our target protein stability with no loss of activity. © 2010 Springer Science+Business Media, LLC.
语种:
英文
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