Developmental defect of cytochrome oxidase mutants of Streptomyces coelicolor A3(2)

Fujimoto, Masahiro; Chijiwa, Masato; Nishiyama, Tatsuya; Takano, Hideaki; Uéda, Kenji

doi:10.1099/mic.0.000332

Cited by 12 publications

(17 citation statements)

References 29 publications

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“…Phosphorylated regulator affects or regulates the transcriptional level of three downstream genes. This resulted in a transcriptional activation of SACE_0528 and SACE_1321 and inactivation of SACE_2475 respiration that transfers electrons from menaquinol to oxygen, requires Cu 2+ to enhance its activity (Fujimoto et al 2016). In the case of WT-Δ0101 strain, its system most likely allows higher intracellular copper ion concentration, making oxidative phosphorylation more active thus generating more energy.…”

Section: Discussionmentioning

confidence: 99%

A two-component system gene SACE_0101 regulates copper homeostasis in Saccharopolyspora erythraea

Qiao

et al. 2020

Bioresour. Bioprocess.

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Background: Saccharopolyspora erythraea (S. erythraea) is a Gram-positive bacterium widely used for the production of erythromycin, a potent macrolide antibiotic. However, the mechanism behind erythromycin production is poorly understood. In the high erythromycin-producer strain S. erythraea HL3168 E3, the level of copper ions positively correlates with erythromycin production. To explain this correlation, we performed a genome-based comparison between the wild-type strain NRRL23338 and the mutant strain HL3168 E3, and further characterized the identified gene(s) by targeted genome editing, mRNA transcript analysis, and functional analysis. Results:The response regulator of the two-component system (TCS) encoded by the gene SACE_0101 in S. erythraea showed high similarity with CopR of TCS CopRS in Streptomyces coelicolor, which is involved in the regulation of copper metabolism. The deletion of SACE_0101 was beneficial for erythromycin synthesis most likely by causing changes in the intracellular copper homeostasis, leading to enhanced erythromycin production. In addition, Cu 2+ supplementation and gene expression analysis suggested that SACE_0101 may be involved in the regulation of copper homeostasis and erythromycin production. Conclusions:The mutation of SACE_0101 gene increased the yield of erythromycin, especially upon the addition of copper ions. Therefore, the two-component system gene SACE_0101 plays a crucial role in regulating copper homeostasis and erythromycin synthesis in S. erythraea.

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Section: Discussionmentioning

confidence: 99%

A two-component system gene SACE_0101 regulates copper homeostasis in Saccharopolyspora erythraea

Qiao

et al. 2020

Bioresour. Bioprocess.

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“…The loss of the bcc-aa 3 respiratory branch leads to moderate to severe growth phenotypes in C. glutamicum, while Mycobacterium tuberculosis cannot grow without this branch of the respiratory chain. A recent study (26) has shown that deletions in the genes encoding the bcc-aa 3 supercomplex led to a developmental phenotype in S. coelicolor on certain growth media (26). However, no studies have been undertaken to examine the consequences to the physiology of Streptomyces coelicolor caused by deleting the genes encoding the cytochrome bd oxidase.…”

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Cytochrome bd Oxidase Has an Important Role in Sustaining Growth and Development of Streptomyces coelicolor A3(2) under Oxygen-Limiting Conditions

et al. 2018

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A3(2) is a filamentously growing, spore-forming, obligately aerobic actinobacterium that uses both a copper -type cytochrome oxidase and a cytochrome oxidase to respire oxygen. Using defined knockout mutants, we demonstrated that either of these terminal oxidases was capable of allowing the bacterium to grow and complete its developmental cycle. The genes encoding the complex and the oxidase are clustered at a single locus. Using Western blot analyses, we showed that the oxidase branch is more prevalent in spores than the oxidase. The level of the catalytic subunit, CydA, of the oxidase was low in spore extracts derived from the wild type, but it was upregulated in a mutant lacking the supercomplex. This indicates that cytochrome oxidase can compensate for the lack of the other respiratory branch. Components of both oxidases were abundant in growing mycelium. Growth studies in liquid medium revealed that a mutant lacking the oxidase branch grew approximately half as fast as the wild type, while the oxygen reduction rate of the mutant remained close to that of the wild type, indicating that the oxidase was mainly functioning in controlling electron flux. Developmental defects were observed for a mutant lacking the cytochrome oxidase during growth on buffered rich medium plates with glucose as the energy substrate. Evidence based on using the redox-cycling dye methylene blue suggested that cytochrome oxidase is essential for the bacterium to grow and complete its developmental cycle under oxygen limitation. Respiring with oxygen is an efficient means of conserving energy in biological systems. The spore-forming, filamentous actinobacterium grows only aerobically, synthesizing two enzyme complexes for O reduction, the cytochrome cytochrome oxidase supercomplex and the cytochrome oxidase. We show in this study that the bacterium can survive with either of these respiratory pathways to oxygen. Immunological studies indicate that the oxidase is the main oxidase present in spores, but the oxidase compensates if the oxidase is inactivated. Both oxidases are active in mycelia. Growth conditions were identified, revealing that cytochrome oxidase is essential for aerial hypha formation and sporulation, and this was linked to an important role of the enzyme under oxygen-limiting conditions.

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“…For example, while deletion of the complete locus shown in Figure 1 restricts, but does not abolish, aerobic growth of the bacterium [Fujimoto et al, 2016;Fischer et al, 2018], the resulting oxidase-negative phenotype of the mutant (strain COE192) can only be restored to an oxidase-positive phenotype when the complete qcr-cta gene locus is supplied on an integrative plasmid [Fischer et al, 2018]. Two genes within this locus, SCO2152 and SCO2153 (Fig.…”

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Cytochrome bcc-aa3 Oxidase Supercomplexes in the Aerobic Respiratory Chain of Streptomyces coelicolor A3(2)

et al. 2018

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Streptomyces coelicolor A3(2), an obligately aerobic, oxidase-positive, and filamentous soil bacterium, lacks a soluble cytochrome c in its respiratory chain, having instead a membrane-associated diheme c-type cytochrome, QcrC. This necessitates complex formation to allow electron transfer between the cytochrome bcc and aa3 oxidase respiratory complexes. Combining genetic complementation studies with in-gel cytochrome oxidase activity staining, we demonstrate that the complete qcrCAB-ctaCDFE gene locus on the chromosome, encoding, respectively, the bcc and aa3 complexes, is required to manifest a cytochrome oxidase enzyme activity in both spores and mycelium of a qcr-cta deletion mutant. Blue-native-PAGE identified a cytochrome aa3 oxidase complex of approximately 270 kDa, which catalyzed oxygen-dependent diaminobenzidine oxidation without the requirement for exogenously supplied cytochrome c, indicating association with QcrC. Furthermore, higher molecular mass complexes were identified upon addition of soluble cytochrome c, suggesting the supercomplex is unstable and readily dissociates into subcomplexes lacking QcrC. Immunological and mass spectrometric analyses of active, high-molecular mass oxidase-containing complexes separated by clear-native PAGE identified key subunits of both the bcc complex and the aa3 oxidase, supporting supercomplex formation. Our data also indicate that the cytochrome b QcrB of the bcc complex is less abundant in spores compared with mycelium.

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Developmental defect of cytochrome oxidase mutants of Streptomyces coelicolor A3(2)

Cited by 12 publications

References 29 publications

A two-component system gene SACE_0101 regulates copper homeostasis in Saccharopolyspora erythraea

A two-component system gene SACE_0101 regulates copper homeostasis in Saccharopolyspora erythraea

Cytochrome bd Oxidase Has an Important Role in Sustaining Growth and Development of Streptomyces coelicolor A3(2) under Oxygen-Limiting Conditions

Cytochrome <b><i>bcc-aa3</i></b> Oxidase Supercomplexes in the Aerobic Respiratory Chain of <b><i>Streptomyces coelicolor</i></b> A3(2)

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