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Cold‐adapted maturation of thermophilic WF146 protease by mimicking the propeptide binding interactions of psychrophilic subtilisin S41
Author(s) -
Yang Yi-Ran,
Zhu Hui,
Fang Nan,
Liang Xiaoliang,
Zhong Chuan-Qi,
Tang Xiao-Feng,
Shen Ping,
Tang Bing
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.06.041
Subject(s) - psychrophile , subtilisin , protease , thermophile , protein precursor , biochemistry , amino acid residue , chemistry , enzyme , amino acid , mutant , biology , peptide sequence , gene
Thermophilic WF146 protease matures efficiently at 60 °C, but quite slowly at low temperatures. In this report, seven amino acid residues involved in interactions between the mature domain and the propeptide of the enzyme were substituted by corresponding residues of psychrophilic subtilisin S41 to generate mutant Mut7 (S105G/G107D/Y117E/S136N/V143G/K144E/D145S). Mut3 (S105G/G107D/Y117E) and Mut4 (S136N/V143G/K144E/D145S) were also constructed. Transferring structural features from S41 endowed Mut7 with a remarkably increased maturation rate, as well as an improved caseinolytic activity at 25 °C. Moreover, Mut3 and Mut4 each obtained one of the above endowments. Further studies suggest that low‐temperature activity and maturation rate are not necessarily linked, and uncoupling structural elements modulating the two properties may be advantageous to cold adaptation.