• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.162-173
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• 2015

The cellular fatty acid composition is important for metabolic plasticity in Rhodobacter sphaeroides. We explored the effects of changing the cellular ratio of unsaturated fatty acids (UFAs) to saturated fatty acids (SFAs) in R. sphaeroides by overexpressing several key fatty acid biosynthetic enzymes through the use of expression plasmid pRK415. Bacteria containing the plasmid pRKfabI₁ with the fabI₁ gene that encodes enoyl-acyl carrier protein (ACP) reductase showed a reduction in the cellular UFA to SFA ratio from 4 (80% UFA) to 2 (65% UFA) and had decreased membrane fluidity and reduced cell growth. Additionally, the ratio of UFA to SFA of the chromatophore vesicles from pRKfabI₁-containing cells was similarly lowered, and the cell had decreased levels of light-harvesting complexes, but no change in intracytoplasmic membrane (ICM) content or photosynthetic (PS) gene expression. Both inhibition of enoyl-ACP reductase with diazaborine and addition of exogenous UFA restored membrane fluidity, cell growth, and the UFA to SFA ratio to wild-type levels in this strain. R. sphaeroides containing the pRKfabB plasmid with the fabB gene that encodes the enzyme β-ketoacyl-ACP synthase I exhibited an increased UFA to SFA ratio from 4 (80% UFA) to 9 (90% UFA), but showed no change in membrane fluidity or growth rate relative to control cells. Thus, membrane fluidity in R. sphaeroides remains fairly unchanged when membrane UFA levels are between 80% and 90%, whereas membrane fluidity, cell growth, and cellular composition are affected when UFA levels are below 80%.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.1031-1037
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• 2001

By a sequential action of ${\beta}$-ketothiolase and acetoacetyl-CoA reductase, two molecules of acetyl-CoA re converted into D-3-hydroxybutyryl-CoA, a substrate for PHB synthase to form poly-3-hydroxybutyryl-CoA, a substrate for PHB synthase to form poly-3-hydroxybutyrate (PHB) of rhodobacter sphaeroides. The ${\beta}$-ketothiolase gene, phbA, and acetoacetyl-CoA reductase gene, phbB, were cloned and analyzed for their expression. Enzyme activities of ${\beta}$-ketothiolase and acetoacetyl-CoA reductase showed constitutive levels during aerobic and photoheterotrophic growth of R. sphaeroides. In addition, no difference of each enzyme activity was observed between cells grown aerobically and photoheterotrophically. The constitutive level of the enzyme activities are regulated according to the growth phases along with growth conditions. Thus, phbAB expression is not determinative in regulating the PB content. On the other hand, phbA-deleted cell AZI accumulated only $10\%$ PHB of the wild-type, and an elevated dosage of phbAB in trans in R. sphaeroides resulted in a higher content of PHB, indicating that phbAB codes for the enzymes responsible for providing the main supply of subsyrate for PHB synthase. PHB formation by an alternative pathway that does not does not depend on the phbA-and phbB-coding enzymes is also proposed.

• Journal of Applied Biological Chemistry
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• v.64 no.3
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• pp.217-223
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• 2021

5-aminolevulinic acid (ALA) is a representative photosensitizer used in numerous fields including cancer diagnosis and treatment. In this study, experiments were conducted to optimize the growth of Rhodobacter sphaeroides and production of ALA through LED irradiation of various wavelengths, addition of organic acid precursors of ALA, and changes in glucose concentration. After 72 h cultivation, the 850 nm wavelength LED irradiated at the same light intensity as the incandescent lamp increased the growth of R. sphaeroides and the production of ALA about 1.5- and 1.8-fold as compared with the control, respectively (p <0.0001 and p <0.0001). As a result of culturing R. sphaeroides by irradiating an LED with a wavelength of 850 nm after adding organic acid to the final concentration of 5 mM in culture medium, the production of ALA was increased about 2.8-fold in medium supplemented with pyruvic acid compared with the control (p <0.0001). In addition, the growth of the strain and the production of ALA were increased about 2.9- and 3.4-fold in medium supplemented with 40 mM glucose compared to the control which added only 5 mM pyruvic acid, respectively (p <0.0001 and p <0.0001). The yield of ALA per cell dry mass was about 1.4 folds higher than that of the control in 20 and 40 mM glucose, respectively (p <0.001). In conclusion, the growth of R. sphaeroides and production of ALA were increased by 850 nm wavelength LED irradiation. It also optimized the growth of R. sphaeroides and production of ALA through organic acid addition and glucose concentration changes.

• Proceedings of the Korean Biophysical Society Conference
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• 2002.06b
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• pp.51-51
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• 2002

PpsR from the facultative photohetrotroph Rhodobacter sphaeroides is involved in repression of photo system gene expression. SDS-PAGE analysis showed that some portion of PpsR is oxidized so that intra- or inter-disulfide bond is formed between the two cysteins in each subunit. The disulfide bond was reduced by dithiothreitol and the binding activity to puc promoter region was increased.(omitted)

• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.219-227
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• 2008

The free-living photoheterotrophic Gram-negative bacterium Rhodobacter sphaeroides possesses a quorum-sensing (QS) regulatory system mediated by CerR-CerI, a member of the LuxR-LuxI family. To identify the genes affected by the regulatory system, random lacZ fusions were generated in the genome of R. sphaeroides strain 2.4.1 using a promoter-trapping vector, pSG2. About 20,000 clones were screened and 23 showed a significantly different level of ${\beta}$-gal activities upon the addition of synthetic 7,8-cis-N-tetradecenoyl-homoserine lactone (RAI). Among these 23 clones, the clone showing the highest level of induction was selected for further study, where about a ten-fold increase of ${\beta}$-gal activity was exhibited in the presence of RAI and induction was shown to be required for cerR. In this clone, the lacZ reporter was inserted in a putative gene that exhibited a low homology with catD. A genetic analysis showed that the expression of the catD homolog was initiated from a promoter of another gene present upstream of the catD. This upstream gene showed a strong homology with luxR and hence was named qsrR (quorum-sensing regulation regulator). A comparison of the total protein expression profiles for the wild-type cells and qsrR-null mutant cells using two-dimensional gel electrophoresis and a MALDI-TOF analysis allowed the identification of sets of genes modulated by the luxR homolog.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.310-316
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• 2001

Poly-3-hydroxybutyrate (PHB) synthase catalyzed the last enzymic step to synthesize the intracellular PHB of Rhodobacter sphaeroides. No PHB was detected when the phbC coding for PhB synthase was interrupted, and its expression was regulated at the level of transcription. The cellular PHB content increased about four- to six-fold during the growth transition from the exponential to the early stationary phase under both aerobic and photoheterotrophic conditions. The PHB content during the aerobic growth seemed to be determined by the PhB synthase activity. However, the PHB synthase activity of photoheterotrophically grown cells did not correlate with the PhB content, suggesting a photoheterotrophic regulation different from the aerobic control. Thus, the PHB content of R. sphaeroides was regulated at the transcription level only under aerobic conditions.

• Fisheries and Aquatic Sciences
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• v.11 no.3
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• pp.172-176
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• 2008

To develop a potent strain for the production of coenzyme $Q_{10}$, a photosynthetic bacterium was isolated from silt of the Nakdong River in Korea. Using l6S-rDNA sequence analysis, the isolated strain was identified as Rhodobacter sphaeroides. A stable improvement in its $CoQ_{10}$ content was achieved by chemical mutation, upon which the content of $CoQ_{10}$(2.94 mg/g dry cell) was increased by approximately 1.9-fold, comparable to that of R. sphaeroides reported in other studies. The isolate is a potentially valuable microorganism for mass production of $CoQ_{10}$, and may provide an appropriate model for further study of economical mass production.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1589-1598
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• 2015

In this work, the anti-aging skin effects of bacteriochlorophyll a isolated from Rhodobacter sphaeroides are first reported, with notably low cytotoxicity in the range of 1% to 14% in adding 0.00078 (% (w/w)) of the extracts, compared with the normal growth of both human dermal fibroblast and keratinocyte cells without any treatment as a control. The highest production of procollagen from human fibroblast cells (CCD-986sk) was observed as 221.7 ng/ml with 0.001 (% (w/w)) of bacteriochlorophyll a, whereas 150 and 200 ng/ml of procollagen production resulted from addition of 0.001 (% (w/w)) of the photosynthetic bacteria. The bacteriochlorophyll-a-induced TNF-α production increased to 63.8%, which was lower secretion from HaCaT cells than that from addition of 0.00005 (% (w/w)) of bacteriochlorophyll a. Additionally, bacteriochlorophyll a upregulated the expression of genes related to skin anti-aging (i.e., keratin 10, involucrin, transglutaminase-1, and MMPs), by up to 4-15 times those of the control. However, crude extracts from R. sphaeroides did not enhance the expression level of these genes. Bacteriochlorophyll a showed higher antioxidant activity of 63.8% in DPPH free radical scavenging than those of water, ethanol, and 70% ethanol extracts (14.0%, 57.2%, and 12.6%, respectively). It was also shown that the high antioxidant activity could be attributed to the skin anti-aging effect of bacteriochlorophyll a, although R. sphaeroides itself would not exhibit significant anti-aging activities.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.211-216
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• 1983

Rhodopseudomonas sphaeroides K-7 evolves large quantities of molecular hydrogen under anaerobic and light illuminated conditions in the presence of utilizable organic compounds as electron donors. Photoevolution of molecular hydrogen was strictly dependent on light as the activity of nitrogenase in this organism. Both of these were inhibited to the nearly same extent at varying concentrations of ammonium ion which also depressed nitrogenase synthesis. In the reaction mixtures devoid of molybdenum ion which is known as the component of nitrogenase, hydrogen evolution also decreased similarly like nitrogenase activity. Photoevolution of molecular hydrogen appeared to have no relationship with hydrogenase activity and bacteriocholophyll content and it was markedly inhibited under the atmosphere of $C_2$H$_2$, $N_2$ or $O_2$. The results strongly indicate that hydrogen evolution by R. sphaeroides K-7 might be catalyzed by nitrogenase. Both hydrogen evolution and nitrogenase activity were largely influenced by the nutritional history of the resting cells. From which we propose that glutamate might play an important role in the regulation of nigrogenase activity in vivo.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.400-406
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• 2012

To improve the hydrogen yield from biological fermentation of organic wastewater, a co-culture system of dark- and photo-fermentation bacteria was investigated. In a pure-culture system of the dark-fermentation bacterium Clostridium butyricum, a pH of 6.25 was found to be optimal, resulting in a hydrogen production rate of 18.7 ml-$H_2/l/h$. On the other hand, the photosynthetic bacterium Rhodobacter sphaeroides could produce the most hydrogen at 1.81mol-$H_2/mol$-glucose at pH 7.0. The maximum specific growth rate of R. sphaeroides was determined to be 2.93 $h^{-1}$ when acetic acid was used as the carbon source, a result that was significantly higher than that obtained using either glucose or a mixture of volatile fatty acids (VFAs). Acetic acid best supported R. sphaeroides cell growth but not hydrogen production. In the co-culture system with glucose, hydrogen could be steadily produced without any lag phase. There were distinguishable inflection points in a plot of accumulated hydrogen over time, resulting from the dynamic production or consumption of VFAs by the interaction between the dark- and photo-fermentation bacteria. Lastly, the hydrogen production rate of a repeated fed-batch run was 15.9 ml-$H_2/l/h$, which was achievable in a sustainable manner.