NISCAIR Online Periodicals Repository

Research Journals >
Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.43 [2006] >
IJBB Vol.43(6) [December 2006] >

Title: Overexpression of a recombinant -glutamyltranspeptidase from
Authors: Yao, Ya-Feng
Weng, Yih-Ming
Hu, Hui-Yu
Lin, Long-Liu
Keywords: Escherichia coli
Signal sequence
Nickel-chelate chromatography
Issue Date: 2006
Publisher: CSIR
Abstract: A truncated Escherichia coli Novablue -glutamyltranspeptidase (EcGGT) gene, lacking the first 48-bp coding sequence for part of the signal sequence, was amplified by polymerase chain reaction (PCR) and cloned into expression vector pQE-30 to generate pQE-EcGGT. The maximum production of His6-tagged enzyme by E. coli M15 (pQE-EcGGT) was achieved with 0.1 mM IPTG induction for 12 h at 20°C. The overexpressed enzyme was purified to homogeneity by nickel-chelate chromatography to a specific transpeptidase activity of 4.25 U/mg protein and a final yield of 83%. The molecular masses of the subunits of the purified enzyme were determined to be 41 and 21 kDa respectively by SDS-PAGE, indicating the precursor EcGGT still undergoes the post-translational processing even in the truncation of signal sequence. His6-tagged EcGGT migrated relative to the molecular mass of approximately 120 kDa and its heterodimeric structure was confirmed by a native-PAGE gel.
Page(s): 345-350
ISSN: 0301-1208
Source:IJBB Vol.43(6) [December 2006]

Files in This Item:

File Description SizeFormat
IJBB 43(6) 345-350.pdf749.13 kBAdobe PDFView/Open
 Current Page Visits: 74 
Recommend this item


Online Submission of Articles |  NISCAIR Website |  National Knowledge Resources Consortium |  Contact us |  Feedback

Disclaimer: NISCAIR assumes no responsibility for the statements and opinions advanced by contributors. The editorial staff in its work of examining papers received for publication is helped, in an honorary capacity, by many distinguished engineers and scientists.

CC License Except where otherwise noted, the Articles on this site are licensed under Creative Commons License: CC Attribution-Noncommercial-No Derivative Works 2.5 India

Copyright © 2015 The Council of Scientific and Industrial Research, New Delhi. All rights reserved.

Powered by DSpace Copyright © 2002-2007 MIT and Hewlett-Packard | Compliant to OAI-PMH V 2.0

Home Page Total Visits: 165288 since 01-Sep-2015  Last updated on 27-Jun-2016Webmaster: