Table of Contents
ISRN Zoology
Volume 2011, Article ID 835274, 5 pages
http://dx.doi.org/10.5402/2011/835274
Research Article

Expression of the Lactate Dehydrogenase Gene from Eptatretus okinoseanus in Escherichia coli

1Faculty of Pharmaceutical Sciences, Toho University, Chiba 2748510, Japan
2Faculty of Science, Toho University, Chiba 2748510, Japan

Received 31 March 2011; Accepted 3 May 2011

Academic Editors: V. Almeida-Val and A. Ramirez-Bautista

Copyright © 2011 Yoshikazu Nishiguchi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Amplified Eptatretus okinoseanus cDNA was digested with NdeI and EcoRI, cloned into pCold trigger factor (TF), and transformed with Escherichia coli strain BL 21 in which a csp A promoter was introduced to inhibit the expression of foreign peptides. Recombinant lactate dehydrogenase (LDH) was obtained in the soluble fraction after sonication of the cells. The protein was digested by HRC 3C protease, thrombin, and factor Xa. The specific activity of TF-tagged protein and tagless protein were 0 . 6 4 6 × 1 0 6  mIU/mg and 3 . 5 6 × 1 0 6  mIU/mg, respectively. The deletion of the TF tag enhanced the activity compared with the native protein to 1 3 . 4 × 1 0 6  mIU/mg, showing that this expression method is effective for the mass production of the protein to allow further study of the structure of LDH.