Successful expression of the Bordetella petrii nitrile hydratase activator P14K and the unnecessary role of Ser115.

ABSTRACT:

Background

The activator P14K is necessary for the activation of nitrile hydratase (NHase). However, it is hard to be expressed heterogeneously. Although an N-terminal strep tagged P14K could be successfully expressed from Pseudomonas putida, various strategies for the over-expression of P14K are needed to facilitate further application of NHase.

Results

P14K was successfully expressed through fusing a his tag (his-P14K), and was over-expressed through fusing a gst tag (gst-P14K) at its N-terminus in the NHase of Bordetella petrii DSM 12804. The stability of gst-P14K was demonstrated to be higher than that of the his-P14K. In addition, the Ser115 in the characteristic motif CXLC-Ser115-C of the active center of NHase was found to be unnecessary for NHase maturation.

Concl

SUBMITTER: Sun W 

PROVIDER: S-EPMC4761151 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature