Expression and production of soluble Mycobacterium tuberculosis H37Rv mycosin-3

Zhuo Fang; Wolf-Dieter Schubert; Nicolaas Claudius Gey van Pittius

doi:10.1016/j.bbrep.2016.02.005

Expression and production of soluble Mycobacterium tuberculosis H37Rv mycosin-3

Biochem Biophys Rep. 2016 Feb 6:5:448-452. doi: 10.1016/j.bbrep.2016.02.005. eCollection 2016 Mar.

Authors

Zhuo Fang¹, Wolf-Dieter Schubert², Nicolaas Claudius Gey van Pittius¹

Affiliations

¹ DST/NRF Centre of Excellence in Biomedical Tuberculosis Research, US/MRC Centre for Molecular and Cellular Biology, Division of Molecular Biology and Human Genetics, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University of Stellenbosch, Francie van Zijl Drive, Tygerberg 7505, South Africa.
² Department of Biochemistry, University of Pretoria, cnr Lynnwood Road and Roper Street, Hatfield, Pretoria 0028, South Africa.

Abstract

Mycobacteria encode five type VII secretion system (T7SS) or ESX for nutrient acquisition and virulence. Mycosins are membrane-anchored components of ESX with serine protease activity but an unidentified substrate range. Establishing the substrate specificity of individual mycosins will help to elucidate individual ESX functions. Mycosin-1 and -3 orthologues from two environmental mycobacterial species, Mycobacterium smegmatis and Mycobacterium thermoresistibile, have been heterologously produced, but mycosins from Mycobacterium tuberculosis (Mtb) remain to be studied. Here we describe the successful production of Mtb mycosin-3 as a first step in investigating its structure and function.

Keywords: Cloning; ESX; Mycobacterium tuberculosis; Mycosin-3; Protein production.