Open AccessEffect of flexibility and positive charge of the C‐terminal domain on the activator P14K function for nitrile hydratase in P seudomonas putida
Author(s) -
Liu Yi,
Cui Wenjing,
Liu Zhongmei,
Cui Youtian,
Xia Yuanyuan,
Kobayashi Michihiko,
Zhou Zhemin
Publication year - 2014
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/1574-6968.12376
Subject(s) - nitrile hydratase , nitrile , pseudomonas putida , chemistry , cobalt , biochemistry , stereochemistry , enzyme , organic chemistry
A self‐subunit swapping chaperone is crucial for cobalt incorporation into nitrile hydratase. However, further information about its structural features is not available. The flexibility and positive charge of the C‐terminal domain of the self‐subunit swapping chaperone (P14K) of nitrile hydratase from P seudomonas putida NRRL ‐18668 play an important role in cobalt incorporation. C‐terminal domain truncation, alternation of C‐terminal domain flexibility through mutant P14K(G86I), and elimination of the positive charge in the C‐terminal domain sharply affected nitrile hydratase cobalt content and activity. The flexible, positively charged C‐terminal domain most likely carries out an external action that allows a cobalt‐free nitrile hydratase to overcome an energetic barrier, resulting in a cobalt‐containing nitrile hydratase.