Streptomyces as host for recombinant production of Mycobacterium tuberculosis proteins.
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.
Vallin
Carlos
C
Department of Biomedical Research, Center of Pharmaceutical Chemistry, Atabey, Playa, Havana, Cuba. val@infomed.sld.cu
Ramos
Astrid
A
Pimienta
Elsa
E
Rodríguez
Caridad
C
Hernández
Tairí
T
Hernández
Ivones
I
Del Sol
Ricardo
R
Rosabal
Grisel
G
Van Mellaert
Lieve
L
Anné
Jozef
J
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000276:immunology; D049109:Cell Proliferation; D003470:Culture Media; D005240:Feasibility Studies; D005285:Fermentation; D007114:Immunization; D008213:Lymphocyte Activation / Q000276:immunology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000276:immunology / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D048170:Streptomyces lividans / Q000378:metabolism*
D001426:Bacterial Proteins; D003470:Culture Media; D011994:Recombinant Proteins