• KSBB Journal
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• v.5 no.3
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• pp.269-277
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• 1990

Methylobacillus sp. SK1, an obligate methylotroph, was cultivated in a fed-batch culture using DO as a methanol feeding indicator with a micro computer-aided control system. While 2.07g/l of cell density was obtained after 13 hr in the batch culture (initial methanol concentration: 1.0%(v/v)),45.3g/l of cell density was obtained after 17 hr by feeding methanol and metal ions in the fed-batch culture with oxygen supply. The high-density biomass was obtained in short cultuivation time by fed-batch culture with feedback control, and consequently the biomass productivity was significantly increased. It was mainly due to extension of logarithmic growth period by methanol feeding without methanol inhibition and intensive aeration without DO limitation with microcomputer-aided control system.

• Korean Journal of Microbiology
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• v.29 no.6
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• pp.380-386
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• 1991

Three types of soluble cytochrome c were purified to homogeneity from Methylobacillus sp. strain SK1 which grows only on methanol. Cytochrome c-I was purified 58.5-fold in seven steps. Cytochrome c-II and c-III were purified 57.3- and 122.1-fold in eight steps, respectively. The molecular weights of the cytochrome c-I was determined to be 12,500, while those of the cytochrome c-II and c-III were 16,000. The isoelectric points of the c-I, c-II and c-III were found to be 8.8, 6.6, and 6.6 respectively. The spectrum of reduced cytochrome c-I showed .alpha.-, .betha.-, .gamma.-peaks at 551.4, 522.2, and 416.6nm. The peaks for c-II were found at 551.0, 521.6, and 416.5nm, while those for c-III were shown at 551.2, 521.8, and 416.0 nm. The spectra of oxidized cytochrome c-I, c-II, and c-III showed .gamma.-peak at 411.8, 409.0, and 410.2 nm, respectively. The absorption coefficients of .alpha.- and .gamma.-peak for c-I in the reduced state were determined as 47 and 197 $mM^{-1}$ $cm^{-1}$ , respectively. The coefficients of .alpha.- and .gamma.-peak for c-II were determined to be 43 and 137 $mM^{-1}$ $cm^{-1}$ , while those for c-III were 41 and 172 $mM^{-1}$ $cm^{-1}$ , respectively. The c-I and c-III were found to bind carbon monoxide.

• Applied Biological Chemistry
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• v.34 no.4
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• pp.386-392
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• 1991

Whole cells of Methylosinus trichosporium OB3b, the obligate methylotroph, were used to produce propylene oxide from propane. This strain has methane monooxygenase, which catalyzes the conversion methane to methanol and can catalyze also the conversion propane to propylene oxide. Optimal condition for the production of propylene oxide was investigated in resting-whole cell system. The optimal pH and temperature was 7.0 and $35^{\circ}C$ respectively. The end product, propylene oxide, didn't inhibit the production of propylene oxide and was not further metabolized in reaction mixture. The addition of methane metabolites (methanol, formaldehde and formic acid) to the reaction mixture stimulated formation of propylene oxide by $3{\sim}4$ times, and methanol was the most effective especially. Under the optimal conditions, the 14.2 mM of propylene oxide was produced after incubation of 60 min. and the conversion ratio of propane to propylene oxide was approximately 8%.

• Korean Journal of Microbiology
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• v.27 no.1
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• pp.63-69
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• 1989

A pink-pigmented facultative methylotrophic bacterium, Methylobacterium sp. strain SY1, was isolated from soil through methanol-enrichment culture technique. The isolate was gram-negative, slightly curved rod, and motile by a single polarly inserted flagellum. The colony was smooth, bright pink, and slimy. The guanine plus cytosine content of the KNA was 66%. The cell was obigately aerobic and exhibited both catalase and oxidase activities. Carotenoid pigment and poly-$\beta$-hydroxybutyrate were present. It was found to have three kinds of plasmid with molecular weights 45,000, 38,500 and 23,000. Growth with methanol(0.5%) was fast ($t_{d}$=6.5h) and was optimal at $30^{\circ}C$ and at pH 7.0. The isolate could grow on several sugars, organic acids, amino acids, amines, and alcohols in addition to the methanol. Methanol was found to be assimilated through the serine pathway.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.207-211
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• 1998

Melhylovorus sp. strain SS1 grown on methanol was found to show activities of key enzymes of the linear route, $NAD^+$-linked formaldehyde and formate dehydrogenases, and the cyclic route, hexulose-6-phosphate synthase, glucose-6-phosphate isomerase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase, for formaldehyde oxidation. The activities of the cyclic route enzymes were higher than those of the linear route enzymes. The bacterium also exhibited activities of the key enzymes of the ribulose monophosphate and Entner-Doudoroff pathways and transaldolase involved in the formaldehyde assimilation and the enzymes involved in the biosynthesis of extracellular polysaccharide. Cells grown in the presence of 2.3 mM ammonium sulfate were higher in the productivity of extracellular polysaccharide, but lower in the growth yield, than those grown in the presence 7.6 mM ammonium sulfate. The activities of 6-phosphogluconate dehydrogenase, phosphoglucomutase, and UDP-pyrophosphorylase in cells grown under nitrogen-limited condition were higher, but that of 6-phosphogluconate dehydratase/2-keto-3-deoxy-6-phosphogluconate aldolase was lower, than those in cells grown in the presence of sufficient amount of nitrogen source.

• Korean Journal of Food Science and Technology
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• v.19 no.4
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• pp.324-330
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• 1987

A large number of methanol-assimilating yeasts and bacteria were isolated from samples of soil, sewage, decomposed milk and spoiled sweet-radish pickles. Among the yeasts, one strain was selected and identified as a strain of Candida boidinii. In 1% (v/v) methanol Candida boidinii YF-3 grew well and could grow in as much as 5%. This yeast required boitin for grwoth. Maximum growth was observed at $30^{\circ}C$ and pH 6 in a semisynthetic medium. The productivity was 2.72g dry cells per liter in batch culture with 1%(v/v) methanol and the cell yield for methanol was $0.39\;gg^{-1}$. The specific growth rate was $0.11\;h^{-1}$ and the generation time was 6.4 hours. The protein content of the cell was 45.5% and total nucleic acid content was 5.9%. The amino acid profile was as good as FAO standard for food protein.

• Microbiology and Biotechnology Letters
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• v.17 no.4
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• pp.392-396
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• 1989

Methylobacterium organophilum, a facultative methylotroph was cultivated on a methanol as a sole carbon and energy source. The cell growth was affected by the various components of minimal synthetic medium and the medium composition was optimized with 0.5% (v/v) methanol at pH 6.8 and at 3$0^{\circ}C$. The maximum specific growth rate of M. organophilum was achieved to 0.26 hr$^{-1}$ in the optimized medium which has following composition: Methanol, 0.5% (v/v):(NH$_4$)$_2$SO$_4$, 1.0g/l:KH$_2$PO$_4$, 2.13g/l:KH$_2$PO$_4$, 1.305g/ι:MgSO$_4$.7$H_2O$. 45g/l and trace elements (CaCl$_2$.2$H_2O$, 3.3mg:FeSO$_4$.7$H_2O$, 1.3mg:MnSO$_4$.4$H_2O$, 130$\mu\textrm{g}$:ZnSO$_4$.5$H_2O$, 40$\mu\textrm{g}$:Na$_2$MoO$_4$.2$H_2O$, 40$\mu\textrm{g}$:CoCl$_2$.6$H_2O$, 40$\mu\textrm{g}$:H$_3$BO$_3$, 30$\mu\textrm{g}$ per liter). By the limitation of nitrogen and deficiency of Mn$^{+2}$ or Fe$^{+2}$, the cell growth was significantly repressed. Methanol greatly repressed the cell growth and the complete inhibition was observed at concentration above 4% (v/v). In order to overcome the methanol inhibition and to prevent the methanol limitation, intermittent feeding of methanol was conducted by a D.O.-stat technique. PHB production by M. organophilum was stimulated by deficiency of nutrients such as NH$_{4}^{+}$, SO$_{4}^{-2}$, $Mg^{+2}$, $K^{+}$, or PO$_{4}^{-3}$ in the medium. The maximum PHB content was obtained as 58% of dry cell weight under deficiency of potassium ion in the optimized synthetic medium.

• 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.