• Journal of Microbiology and Biotechnology
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• v.19 no.1
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• pp.17-22
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• 2009

The mixotrophic growth with methanol plus thiosulfate was examined in nutrient-limited mixotrophic condition for Methylobacterium goesingense CBMB5 and Methylobacterium fujisawaense CBMB37. Thiosulfate oxidation increased the growth and protein yield in mixotrophic medium that contained 150mM methanol and 20mM sodium thiosulfate, at 144 h. Respirometric study revealed that thiosulfate was the most preferable reduced inorganic sulfur source, followed by sulfite and sulfur. M. goesingense CBMB5 and M. fujisawaense CBMB37 oxidized thiosulfate directly to sulfate, and intermediate products of thiosulfate oxidation such as polythionates, sulfite, and sulfur were not detected in spent medium and they did not yield positive amplification for tested soxB primers. Enzymes of thiosulfate oxidation such as rhodanese and sulfite oxidase activities were detected in cell-free extracts of M. goesingense CBMB5, and M. fujisawaense CBMB37, and thiosulfate oxidase (tetrathionate synthase) activity was not observed. It indicated that both the organisms use the "non-S4 intermediate" sulfur oxidation pathway for thiosulfate oxidation. It is concluded from this study that M. goesingense CBMB5, and M. fujisawaense CBMB37 exhibited mixotrophic metabolism in medium containing methanol plus thiosulfate and that thiosulfate oxidation and the presence of a "Paracoccus sulfur oxidation" (PSO) pathway in methylotrophic bacteria are species dependant.

• Korean Journal of Environmental Agriculture
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• v.30 no.2
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• pp.223-228
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• 2011

BACKGROUND: Rhizosphere bacteria may improve plant growth and productivity both by supply nutrients and hormonal stimulation. Although many experiments have shown improvements in plant growth with inoculation of bacterial cultures to the rhizosphere, the main obstacle in the applications of plant growth promoting rhizobacteria in a large scale is the inconsistency of the results. We tested the growth promoting effects of Azospirillum and Methylobacterium strains on red pepper plant. METHODS AND RESULTS: Red pepper seedlings were grown for 25 days in a growth media inoculated with A. brasilense CW903 or M. oryzae CBMB20. The seedlings were transplanted and grown for 45 days in pots with soil in a greenhouse, at half the recommended level of fertilizer. Bacterial culture, $4.0{\times}10^9$ for A. brasilense CW903 and $5.8{\times}10^8$ CFU for M. oryzae CBMB20, was applied in root zone soil periodically every 10 days during the experiment. Inoculation of M. oryzae CBMB20 significantly increased the red pepper plant growth in terms of leaf number, height and mass of shoot, or root mass compared to uninoculated control plants. Although beneficial effects of A. brasilense on plant growth of many crops were observed, the growthpromoting effect of A. brasilense CW903 on red pepper plant was not found in this study. CONCLUSION(s): The factors responsible for the irregularities in plant growth promoting of rhizobacteria are difficult to elucidate. Extensive inoculation experiments in the greenhouse and in the field should enable us to define the factors critical to obtain successful application of plant growth promoting rhizobacteria.

• Journal of Microbiology and Biotechnology
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• v.8 no.4
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• pp.301-304
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• 1998

Methylobacterium organophilum can use nitrogen in the form of ammonium ions ($($NH_4$)_2SO_4\;and\;NH_4Cl) and from nonammonium sources such as glycine, alanine, peptone, and yeast extract. When potassium was limited, significantly more PHB was produced when the ammonium ion was the nitrogen source rather than a nonammonium form. With ammonium, the amount of PHB produced was 0.50-0.53 g PHB/l or $52.0~53.2\%$ of the dry cell weight. If nitrogen was from a nonammonium source, the respective values were 0.04~0.06 g PHB/1 or $8.1~11.3\%$ of dry cell weight. When ammonium sulfate was the sole source of nitrogen under potassium-limited conditions, cell growth and PHB accumulation increased as the pH increased from 6.0 to 7.5. Cell growth and PHB amount at pH 7.5 were 2.50 g dry cell weight/1 and 1.40 g PHB/1, respectively.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.sup3
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• pp.279-285
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• 2016

It has been established that different kinds of bacteria agents are involved in various cancers. Although the mechanism of tumorigenesis is not clearly understood, there is evidence for the presence of bacteria within tumors, with at least a progression effect for some bacteria that prepare suitable microenvironments for tumor cell growth. The aim of current study was to evaluate bacterial dysbiosis in sentinel lymph nodes of breast cancer patients. One hundred and twenty three fresh-frozen sentinel lymph nodes and a corresponding number of normal adjacent breast tissue specimens and five normal mastectomy samples were investigated employing RT-PCR. In addition using genus-specific primers were applied. There was a significant differences as presence of Methylobacterium radiotolerance DNA recorded between patients and normal control group (p= 0.0). Based on our research work, further studies into the role of microbes in breast cancer would be of great interest.

• KSBB Journal
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• v.6 no.1
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• pp.35-43
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• 1991

The production of poly-$\beta$-hydroxybutyrate(PHB) from methanol by batch and fed-batch cultivations of Methylobacterium sp. GL-10 was studied. PHB accumulation was stimulated by the nutrients deficiency including, NH4+, SO42-, and K+. The nitrogen deficiency was the most critical factor for PHB accumulation. In batch cultivation, the maximum cell concentration and PHB content were 1.86g/l and 0.62g/l, respectively, with 1.0%(v/v) of methanol and 0.5g/1 of ammonium sulfate. The mass doubling time of Methylobacterum sp. GL-10 was in the range of 4-5 hrs. The cell growth and PHB accumulation were severely inhibited at the methanol concentration over than 2% (v/v). To overcome methanol Inhibition, constant feeding and intermittent feedillg fed-batch cultivations were adopted, using C/N molar ratio as a control factor. In constant feeding fed-batch process, cell concentration was increased up to 2.67g/1, and PHB yield was enhanced from 0.33 of batch culture to 0.53. The relatively low cell concentration was caused by methanol accumulated in culture broth at late growth phase. To prevent methanol accumulation and to maximize PHB production, DO-state intermittent fed-batch cultivation was attempted. The cell and PHB concentration was reached up to 4.55g/1 and 1.80g/1, respectively. It was possible to maintain methanol concentration low and also to feed nutrient of desired C/N molar ratio.

• Korean Journal of Microbiology
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• v.29 no.6
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• pp.387-391
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• 1991

Methylobacterium extorquens AM1 growing on methanol as a sole source of carbon and energy was found to grow most rapidly (t$t_{d}$ =6h) at 30.deg.C in a mineral medium (pH 7.0) containing 0.5% (v/v) methanol which was agitated at 200 rpm (optimal cultivation condition). Cells grown under the optimal cultivation condition contained more spermidine, but less putrescine, than the cells grown on 2.5%(v/v) ( $t_{d}$ =8h ) or at 20.deg.C ( $t_{d}$ =8h ). Cells cultivated under the optimal condition was found to contain more spermidine than the cells grown at pH 6.0 (( $t_{d}$ =7h ) and pH 8.0 ($t_{d}$ =7.3h). the cells growing at the stationary phase under the optimal condition accumulated more spermine or putrescine than the cells growing at the same phase on 2.5%(v/v) methanol or at pH 6.0 and pH 8.0, respectively. M. extorquens AM1 grown in a medium agitated at 100 rpm ( $t_{d}$ =8.8h ) contained less spermidine and spermine than the cells grown under the optimal cultivation condition.

• Korean Journal of Soil Science and Fertilizer
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• v.43 no.4
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• pp.506-513
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• 2010

Plant growth promoting ability of Methylobacterium oryzae CBMB20 was evaluated under different levels of organic fertilizer application on red pepper plants in a pot experiment. Oil cake as an organic N fertilizer was applied at the rates of 70, 85, 100 and 120% of the conventional recommended level. Each treatment was further treated with or without M. oryzae CBMB20 inoculation. The recommended amount of compost for red pepper was added in all the treatments. Results revealed that plant height, dry biomass and fruit yield were enhanced in increasing order as the rate of fertilization increased. Overall plant growth was improved due to the inoculation of M. oryzae CBMB20 and red pepper fruit yield was also increased by 10-35% in the plants inoculated with M. oryzae CBMB20 at different rates of organic fert1izer application. Total methylotrophic bacterial population in rhizosphere soil measured at the time of harvest was significantly higher in M. oryzae CBMB20 inoculated treatments. The growth promoting effect of M. oryzae CBMB20 found in red pepper could be due to the effective colonization of the bacteria in the rhizosphere and its ability of enhancing nutrient availability and producing plant growth hormones. With the plant growth promoting effect of M. oryzae CBMB20, the rate of organic fertilizer application can be reduced without any significant decreases in biomass production and yield of red pepper.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.533-538
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• 1992

The generation time of Methylobacterium extorquens AMI growing on methanol at pH 5.5 and 7.0 was found to be 23 hand 8.3 h. respectively. The bacterium grown at pH 7.0 were found to contain more amounts of spermidine and putrescine than the cell grown at pH 5.5. Cells grown at both conditions exhibited strong methanol dehydrogenase (MDH) activity at the mid-exponential growth phase. The amounts of MDH. however. were found to be almost equal through all gro~1h phases. Cells growing at the stationary phase contained large amounts of cytochrome c. The cytochrome c content was higher in cells growing at pH 7.0 than the cells growing at pH 5.5. Cells growing at pH 5.5 in the presence of putrescine or spermidine contained increased amounts of putrescine. The level of spermine, however. was decreased and that of spermidine was not changed. Spermine added into the medium was found to have no effect on the level of cellular polyamines. Putrescine or spermidine added into the medium stimulated MDH and hydroxypyruvate reductase activities. but did not affect the contents of MDH and cytochrome c. It was found that preincubation of cell-free extracts with polyamines does not stimulate MDH and hydroxypyruvate reductase activities.

• Korean Chemical Engineering Research
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• v.55 no.3
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• pp.363-368
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• 2017

Methylotrophy is able to use reduced one-carbon compound, such as methanol and methylamine, as a sole carbon source. Methylobacterium extorquens AM1 is the most extensively studied methylotroph utilizing serine-isocitrate lyase cycle. Because the Poly 3-hydroxybutyrate (PHB) synthesis pathway in M. extorquens AM1 is likely to interlink with EMCP (ethylmalonyl-CoA pathway), glyoxylate, and TCA cycles, regulation of PHB production is needed to produce EMCP-derived acid or TCA acids. To adjust carbon flux to PHB production, PhaR, which seems to have function of regulator of PHB synthesis and acetyl-CoA flux, was knocked out in M. extorquens AM1 by using markerless gene deletion methods. As a result, PHB granules were remarkably reduced in the knockout strain ${\Delta}phaR$ compared to parental strain. Although lag phase was extended for 12h, ${\Delta}phaR$ showed similar cell growth and methanol consumption rate compared to wild type.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.244-249
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• 1998

An extracellular polysaccharide, methylan, was produced under the specific conditions by Methylobacterium organophilum from methanol. The specific growth rate of cells was approximately constant regardless of C/N ratio and the specific product yield was maximum at a C/N ratio of 30. Methylan production was suppressed by the deficiency of mineral ions such as Mn$^{++}$ or Fe$^{++}$ ion. The optimal pH for cell growth and methylan production was 7. Whereas the optimal temperature for cell growth was found to be 37$^{\circ}C$, that for methylan production was 3$0^{\circ}C$. The methanol concentration above 4% completely inhibited the cell growth. The initial methanol concentration for the maximal production of methylan was 0.5% (v/v) and above this concentration, methylan production was markedly inhibited. To overcome the substrate toxicity and inhibition for both cell growth and methylan production, a fed-bach culture of intermittent feeding within 5 g/l methanol was conducted under the optimal culture condition. Methylan production of was stimulated by nitrogen limitation and methylan was accumulated up to 8.7 g/1 and cell mass also increased up to 12.4 g/l.