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Updated: Aug 9, 2026
Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria
Published on: December 8, 2023
Streptomyces as host for recombinant production of Mycobacterium tuberculosis proteins
Carlos Vallin1, Astrid Ramos, Elsa Pimienta
1Department of Biomedical Research, Center of Pharmaceutical Chemistry, Atabey, Playa, Havana, Cuba. val@infomed.sld.cu
Abstract:
The 45/47 kDa APA protein (Rv1860) of Mycobacterium tuberculosis was produced by Streptomyces lividans. The recombinant protein could be recovered from the culture medium of an S. lividans clone containing the apa gene under control of the promoter and signal sequence of the Streptomyces coelicolor agarase gene. The recombinant protein production was further scaled-up using fermentation conditions. The APA protein was subsequently purified from the culture supernatant by means of immunochromatography. About 80 mg of recombinant protein were obtained per liter of culture media. In vivo tests with the APA protein purified from S. lividans TK24/pRGAPA1 revealed that the recombinant protein was antigenic and could induce high titers of specific antibodies in the mouse biological model. Results obtained concerning heterologous production of APA, its immunogenic and antigenic capacity, demonstrated the potential of S. lividans as a valuable host for the production of recombinant proteins from M. tuberculosis.
Insights
This study demonstrates that Streptomyces lividans can effectively produce the Mycobacterium tuberculosis 45/47 kDa APA protein. The recombinant APA protein is antigenic and induces specific antibodies, highlighting its potential for tuberculosis diagnostics.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- The 45/47 kDa APA protein (Rv1860) is a target antigen of Mycobacterium tuberculosis.
- Efficient production of recombinant tuberculosis proteins is crucial for diagnostics and research.
- Heterologous expression systems are being explored for Mycobacterium tuberculosis antigens.
Purpose of the Study:
- To evaluate Streptomyces lividans as a host for the heterologous production of the Mycobacterium tuberculosis 45/47 kDa APA protein.
- To assess the antigenic and immunogenic properties of the recombinant APA protein.
- To determine the potential of S. lividans for large-scale recombinant protein production.
Main Methods:
- The apa gene from Mycobacterium tuberculosis was cloned and expressed in Streptomyces lividans under the control of the Streptomyces coelicolor agarase gene promoter and signal sequence.
- Recombinant protein production was scaled up using fermentation.
- Purification of the APA protein was achieved via immunochromatography.
- In vivo immunogenicity was tested in a mouse model.
Main Results:
- Recombinant 45/47 kDa APA protein was successfully produced and secreted into the culture medium of S. lividans.
- Approximately 80 mg of purified recombinant protein was obtained per liter of culture.
- The recombinant APA protein was shown to be antigenic and induced high titers of specific antibodies in mice.
- The purified protein demonstrated significant immunogenic capacity.
Conclusions:
- Streptomyces lividans is a suitable and valuable host for the heterologous production of the Mycobacterium tuberculosis 45/47 kDa APA protein.
- The recombinant APA protein possesses antigenic and immunogenic properties, indicating its potential utility.
- This study supports the use of S. lividans for producing recombinant proteins from Mycobacterium tuberculosis for further applications.
