Real-time monitoring of rhamnose induction effect on the expression of mpt64 gene fused with pelB signal peptide in Escherichia coli BL21 (DE3)

Sri Agung Fitri Kusuma; Ida Parwati; Toto Subroto; Yaya Rukayadi; Tina Rostinawati; Muhammad Yusuf; Muhammad Fadhlillah; Laily D Tanti; Risa R Ahyudanari

doi:10.4103/japtr.JAPTR_120_19

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ORIGINAL ARTICLE

Year : 2020 | Volume : 11 | Issue : 2 | Page : 69-73

Real-time monitoring of rhamnose induction effect on the expression of mpt64 gene fused with pelB signal peptide in Escherichia coli BL21 (DE3)

Sri Agung Fitri Kusuma¹, Ida Parwati², Toto Subroto³, Yaya Rukayadi⁴, Tina Rostinawati⁵, Muhammad Yusuf³, Muhammad Fadhlillah⁶, Laily D Tanti⁷, Risa R Ahyudanari⁷
¹ Department of Chemistry, Faculty of Mathematics and Natural Sciences; Department of Biology Pharmacy, Faculty of Pharmacy, Padjadjaran University, Bandung, Indonesia
² Department of Clinical Pathology, Faculty of Medical, Padjadjaran University, Bandung, Indonesia
³ Department of Chemistry, Faculty of Mathematics and Natural Sciences; Research Center of Molecular Biotechnology and Bioinformatics, Padjadjaran University, Bandung, Indonesia
⁴ Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang, Malaysia
⁵ Department of Biology Pharmacy, Faculty of Pharmacy, Padjadjaran University, Bandung, Indonesia
⁶ Department of Chemistry, Faculty of Mathematics and Natural Sciences; Research Center of Molecular Biotechnology and Bioinformatics, Padjadjaran University, Bandung; 6PT. GenPro Multiguna Sejahtera, Sumedang, Indonesia
⁷ Department of Chemistry, Faculty of Mathematics and Natural Sciences, Padjadjaran University, Bandung, Indonesia

Correspondence Address:
Prof. Toto Subroto
Jl. Sentral No. 39 Cimahi 50413, West Java
Indonesia

Source of Support: None, Conflict of Interest: None

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DOI: 10.4103/japtr.JAPTR_120_19

In this research, Escherichia coli BL21 (DE3) harboring an expression vector constructed with a rhamnose-inducible promoter and a pelB signal peptide was used as a host cell to produce MPT64 protein. The objective of this research was to figure out the optimum time of mpt64 gene expression through real-time monitoring of MPT64 protein production and distribution in host compartments. The mpt64 expression was regulated by the rhamnose presence at a concentration of 4 mM. The real-time isolated protein was monitored using polyacrylamide gel electrophoresis in denaturation condition. Based on real-time monitoring, the MPT64 protein (24 kDa) in the cytoplasm was optimum detected at 24 h after induction. For periplasmic fraction, the protein was detected at 4 h after induction but thinning at 15 h after induction. At 16 h after induction, the MPT64 protein band was found in the medium with increasing concentrations until 24 h. Thus, it can be concluded that the mpt64 gene expression was regulated in the presence of rhamnose as an inducer, and the proteins were shown to be translocated throughout the host cell compartment with different levels of protein accumulation at different times, according to the role of pelB as a signal peptide.

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