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Updated: Mar 18, 2026
Visual and Microscopic Evaluation of Streptomyces Developmental Mutants
Published on: September 12, 2018
Developmental defect of cytochrome oxidase mutants of Streptomyces coelicolor A3(2)
Masahiro Fujimoto1, Masato Chijiwa1, Tatsuya Nishiyama1
1Life Science Research Center, College of Bioresource Sciences, Nihon University, 1866 Kameino, 252-0880 Fujisawa, Japan.
Abstract:
To study the link between energy metabolism and secondary metabolism/morphological development in Streptomyces, knockout mutants were generated with regard to the subunits of the cytochrome oxidase supercomplex (CcO) in Streptomyces coelicolor A3(2). All mutants exhibited an identical phenotype: viable but defective in antibiotic production and cell differentiation when grown in both complex and minimal media. The growth yield of the CcO mutant was about half of that of the WT strain on glucose medium while both strains grew similarly on maltose medium. Intracellular ATP measurement demonstrated that the CcO mutant exhibited high intracellular ATP level. A similar elevation of intracellular ATP level was observed with regard to the WT strain cultured in the presence of BCDA, a copper-chelating agent. Reverse transcriptase PCR analysis demonstrated that the transcription of ATP synthase operon is upregulated in the CcO mutant. Addition of carbonylcyanide m-chlorophenylhydrazone, an inhibitor of ATP synthesis, promoted antibiotic production and aerial mycelia formation in the CcO mutant and BCDA-treated WT cells. We hypothesize that the deficiency of CcO causes accumulation of intracellular ATP, and that the high ATP level inhibits the onset of development in S. coelicolor.
Insights
Cytochrome oxidase supercomplex (CcO) deficiency in Streptomyces coelicolor halts antibiotic production and cell differentiation by increasing intracellular ATP levels. Inhibiting ATP synthesis restores these developmental processes.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Energy metabolism is crucial for bacterial growth, secondary metabolism, and morphological development.
- Cytochrome oxidase supercomplex (CcO) plays a key role in cellular respiration and energy production.
- Streptomyces coelicolor is a model organism for studying antibiotic biosynthesis and differentiation.
Purpose of the Study:
- To investigate the link between energy metabolism and secondary metabolism/morphological development in Streptomyces.
- To determine the role of CcO in Streptomyces coelicolor A3(2) development.
- To elucidate the impact of CcO deficiency on antibiotic production and cell differentiation.
Main Methods:
- Generation of knockout mutants for CcO subunits in Streptomyces coelicolor A3(2).
- Phenotypic analysis of mutants grown in complex and minimal media.
- Measurement of intracellular ATP levels.
- Reverse transcriptase PCR to analyze ATP synthase operon transcription.
- Treatment with carbonylcyanide m-chlorophenylhydrazone (an ATP synthesis inhibitor).
Main Results:
- CcO mutants were viable but defective in antibiotic production and cell differentiation.
- CcO mutants showed reduced growth yield on glucose but similar growth on maltose compared to WT.
- Intracellular ATP levels were elevated in CcO mutants and in WT cells treated with a copper-chelating agent (BCDA).
- ATP synthase operon transcription was upregulated in CcO mutants.
- Inhibiting ATP synthesis restored antibiotic production and aerial mycelia formation in CcO mutants and BCDA-treated WT cells.
Conclusions:
- Deficiency in CcO leads to the accumulation of intracellular ATP in Streptomyces coelicolor.
- High intracellular ATP levels inhibit antibiotic production and cell differentiation.
- Energy metabolism, specifically ATP levels, directly influences secondary metabolism and morphological development in Streptomyces.
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