Rv1886c Gene Expression for the Production of Mycobacterium tuberculosis Major Secreted Protein Antigen 85B in Escherichia coli

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Napassorn Chamchoi Krittika Rudeejaroonrung Thanchanok Nantamas Ponchanok Suwan Suparwadee Lormlek Varittha Sritalaharauthai Siwarutt Boonyarattanakalin

Abstract

Tuberculosis (TB) is one of the major infectious diseases that cause health problems worldwide. Antigen 85B is a secreted protein of the infectious strain Mycobacterium tuberculosis. Our main focus is on production of proteins as a booster vaccine to replace the traditional Mycobacterium bovis bacillus Calmetter-Guerin (BCG) vaccine formula. The main challenge is to express a high yield of an active recombinant protein in native soluble form.  To the extent of our knowledge, the cultivation conditions, such as optimal temperature, for overexpressing soluble Ag85B protein of the gene Rv1886c, have never been reported. This study showed for the first time the optimizing culturing conditions for inducing expression of soluble Ag85B protein by isopropyl β-D-1-thiogalactopyranoside (IPTG) in Escherichia coli BL21 (DE3) pLysS. Protein yields were higher at a low temperature of 25 °C (for 12 h), compared to those at a high temperature of 37 °C (for 5 h).  To conclude, low temperature is associated with slow expression which allows the protein to adopt a well-folded conformation and provides a high yield of soluble recombinant protein.

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How to Cite
Chamchoi, N., Rudeejaroonrung, K., Nantamas, T., Suwan, P., Lormlek, S., Sritalaharauthai, V., & Boonyarattanakalin, S. (2017). Rv1886c Gene Expression for the Production of Mycobacterium tuberculosis Major Secreted Protein Antigen 85B in Escherichia coli. Science & Technology Asia, 22(1), 19-26. Retrieved from https://www.tci-thaijo.org/index.php/SciTechAsia/article/view/80872
Section
Biological sciences
Author Biographies

Napassorn Chamchoi, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,

Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,

Khlong Luang, Pathum Thani 12120, Thailand

Krittika Rudeejaroonrung, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,

Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,

Khlong Luang, Pathum Thani 12120, Thailand

Thanchanok Nantamas, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,
Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,
Khlong Luang, Pathum Thani 12120, Thailand

Ponchanok Suwan, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,
Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,
Khlong Luang, Pathum Thani 12120, Thailand

Suparwadee Lormlek, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,
Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,
Khlong Luang, Pathum Thani 12120, Thailand

Varittha Sritalaharauthai, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,
Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,
Khlong Luang, Pathum Thani 12120, Thailand

Siwarutt Boonyarattanakalin, School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus, Khlong Luang, Pathum Thani 12120, Thailand

School of Bio-Chemical Engineering and Technology,
Sirindhorn International Institute of Technology, Thammasat University Rangsit Campus,
Khlong Luang, Pathum Thani 12120, Thailand