• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.169-175
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• 2000

The filamentous soil bacterium Streptomyces coelicolor is known to produce four distinct antibiotics. The simultaneous global regulation for the biosynthesis of those four antibiotics was previously confirmed by absA and absB mutations that blocked all four antibiotics' biosynthesis without influencing their morphological differentiation. To study the complex regulatory cascade that controls the secondary metabolism in Streptomyces, a new abs-like mutation was characterized. namely absR, which is slightly leaky on a complete R2YE medium, yet tight on a minimal medium. A genetic analysis of the absR locus indicated that it is located at 10 o'clock on the genetic map, near the site of absA. A cloned copy of the absA gene that encoded bacterial two-component regulatory kinases did not restore antibiotic biosyntheis to the absR mutant. Accordingly, it is proposed that absR is another abs-type mutation which is less tight than the previously identified absA or absB mutations income medium conditions, and can be used to characterize another global regulatory gene for secondary metabolete formation in S. coelicolor.

• Journal of Microbiology and Biotechnology
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• v.31 no.11
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• pp.1591-1600
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• 2021

Streptomyces coelicolor is a filamentous soil bacterium producing several kinds of antibiotics. S. coelicolor abs8752 is an abs (antibiotic synthesis deficient)-type mutation at the absR locus; it is characterized by an incapacity to produce any of the four antibiotics synthesized by its parental strain J1501. A chromosomal DNA fragment from S. coelicolor J1501, capable of complementing the abs- phenotype of the abs8752 mutant, was cloned and analyzed. DNA sequencing revealed that two complete ORFs (SCO6992 and SCO6993) were present in opposite directions in the clone. Introduction of SCO6992 in the mutant strain resulted in a remarkable increase in the production of two pigmented antibiotics, actinorhodin and undecylprodigiosin, in S. coelicolor J1501 and abs8752. However, introduction of SCO6993 did not show any significant difference compared to the control, suggesting that SCO6992 is primarily involved in stimulating the biosynthesis of antibiotics in S. coelicolor. In silico analysis of SCO6992 (359 aa, 39.5 kDa) revealed that sequences homologous to SCO6992 were all annotated as hypothetical proteins. Although a metalloprotease domain with a conserved metal-binding motif was found in SCO6992, the recombinant rSCO6992 did not show any protease activity. Instead, it showed very strong β-glucuronidase activity in an API ZYM assay and toward two artificial substrates, p-nitrophenyl-β-D-glucuronide and AS-BI-β-D-glucuronide. The binding between rSCO6992 and Zn²⁺ was confirmed by circular dichroism spectroscopy. We report for the first time that SCO6992 is a novel protein with β-glucuronidase activity, that has a distinct primary structure and physiological role from those of previously reported β-glucuronidases.