Open AccessCloning, expression, purification and preliminary X‐ray diffraction studies of a putative Mycobacterium smegmatis thiolase
Author(s) -
Janardan Neelanjana,
Paul Anju,
Harijan Rajesh K.,
Wierenga Rikkert K.,
Murthy M. R. N.
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111019324
Subject(s) - mycobacterium smegmatis , thiolase , cloning (programming) , microbiology and biotechnology , gene , biology , chemistry , biochemistry , peroxisome , mycobacterium tuberculosis , medicine , tuberculosis , pathology , computer science , programming language
Thiolases are important in fatty‐acid degradation and biosynthetic pathways. Analysis of the genomic sequence of Mycobacterium smegmatis suggests the presence of several putative thiolase genes. One of these genes appears to code for an SCP‐x protein. Human SCP‐x consists of an N‐terminal domain (referred to as SCP2 thiolase) and a C‐terminal domain (referred as sterol carrier protein 2). Here, the cloning, expression, purification and crystallization of this putative SCP‐x protein from M. smegmatis are reported. The crystals diffracted X‐rays to 2.5 Å resolution and belonged to the triclinic space group P 1. Calculation of rotation functions using X‐ray diffraction data suggests that the protein is likely to possess a hexameric oligomerization with 32 symmetry which has not been observed in the other six known classes of this enzyme.