• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.273-279
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• 1990

For poly- $\beta$ -hydroxybutyrate (PHB) production, a pink-pigmented facultative methylotrophic bacterium (PPFM) P-10 was newly isolated from soils through methanol-enrichment culture. The optimal medium composition for cell growth was 1.0% (vlv) of methanol as carbon source and l.Og/l of ,TEX>$NH_4Cl$, equivalent to C/N ratio of 13.2 at pH 7.0 and $30^{\circ}C$. To investigate the optimal condition for YHB accumulation, two-stage culture technique was adopted; first stage for cell growth and second stage for accumulation of PHB providing unbalanced growth conditions. The optimal PHB accumulation was 1.0% (vIv) of methanol and 0.26gll of $NH_4Cl$, C/N of 50.8 at pH 6.0. To overcome methanol inhibition on cell growth, intermittent feeding fed-batch culture technique was employed, and the cell concentration as high as 14gll with 40% of PHB was achieved. The purified PHB was identified using IR and $1^H NMR$ as homopolymer of 8hydroxybutyric acid. The absorption spectrum of extracted pink colored microbial pigment was alsa investigated.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1978.10a
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• pp.205.3-205
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• 1978

Methanol이용 미생물의 집적배양을 통해 토양 및 하수로부터 분리하여 그 중에서 비교적 생육속도가 빠른 균주를 선별하였다. 이 균주는 형태적, 생리적 특성에 따라 Methylomonas methanolica로 동정되었으며 obligate methylotroph 이었다. 균체 생산량을 높이기 위한 배지조성과 배양 최적조건을 검토한 결과, 탄소원으로는 methanol 0.8%(v/v), 질소원은 $(NH_4)_2SO_4$ 0.6%, 금속이온은 $MgSO_4.$ $7H_2O$ 0.1%이었고, 최적 pH는 6.3, 최적 배양온도는 $32.5^{\circ}C이었으며,$ 생육인자는 요구되지 않았다. 그리고 최적 배양조건에서 1ι용 fer-mentor를 사용하여 회분배양을 하였을 때 최대 비증식속도 0.19$hr^{-1}$, 균체수율은 0.47g dry cell/g-methanol이었다. Chemostat를 이용한 연속배양시 균체생산을 위한 최적희석률은 D=0.1 $hr^{-1}$이었고 이때의 균체생산속도는 0.21g- dry cell/hr이었다. 생산된 건조균체의 단백질과 핵산함량은 각각 73%, 12% 이었다.

• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.244.3-245
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• 2001

No Abstract, See Full Text

• Journal of Microbiology and Biotechnology
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• v.23 no.12
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• pp.1774-1778
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• 2013

Methylotrophs within biological activated carbon (BAC) systems have not received attention although they are a valuable biological resource for degradation of organic pollutants. In this study, methylotrophic populations were monitored for four consecutive seasons in BAC of an actual drinking water plant, using ribosomal tag pyrosequencing. Methylotrophs constituted up to 5.6% of the bacterial community, and the methanotrophs Methylosoma and Methylobacter were most abundant. Community comparison showed that the temperature was an important factor affecting community composition, since it had an impact on the growth of particular methylotrophic genera. These results demonstrated that BAC possesses a substantial methylotrophic activity and harbors the relevant microbes.

• Korean Journal of Microbiology
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• v.28 no.4
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• pp.290-296
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• 1990

Putrescine, spermidine, and spermine were found to persent in Methylobacterium extorquens AM1 growing on methanol, succinate, glucose, or nutrient broth as an energy source. Spermidine was found to be a major polyamine in cells growing on methanol or succinate, while putrescine to be the one in nutrient broth-grown cells. The overall content of polyamines in cells growing on glucose was less than that in cells growing on other substrates. Spermine was the most abundant polyamine in glucose-grown cells. Accumulation of polyamines in M. extorquens AM1 was maximal at the mid-exponential or early stationary phase during growth on each substrate. The effect of polyamines added into the medium on the polyamine composition in M. extorquens AM1 was variable. Each polyamine added into the nutrient broth medium was found to increase the amount of the respective polyamine in the cell. Exogeneously added polyamines had no effect on the growth of M. extorquens AM1.

• 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 Microbiology
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• v.29 no.4
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• pp.250-257
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• 1991

An obligate methanol-oxidizing bacterium, Methylobacillus sp. strain SK1, which grows only on methanol was isolated from soil. The isolate was nonmotile Gram-negtive rod. It does not have internal membrane system. The colonies were small, whitish-yellow, and smooth. The guanine plus cytosine content of the DNA was 48 mol%. Cellular fatty acids consisted predominantly of large amounts of straight-chain saturated $C_{16:0}$ acid and unsaturated $C_{16:1}$ acid. The major ubiquinone was Q-8, and Q-10 was present as minor component. The cell was obligately aerobic and exhibited catalase, but no oxidase, activity. Poly-.betha.-hydroxybutyrate, endospores, or cysts were not observed. the isolate could grow only on methanol in mineral medium. Growth factors were not required. The isolate was unable to use methane, formaldehyde, formate, methylamine, and several other organic compounds tested as a sole source of carbon and energy. Growth was optimal at 35.deg.C and pH 7.5. It could not grow at 42.deg.C. The doubling time was 1.2h at 30.deg.C when grown with 1.0%(v/v) methanol. The growth was not affected by antibiotics inhibiting cell wall synthesis and carbon monoxide but was completely suppressed by those inhibiting protein synthesis. Methanol was found to be assimilated through the ribulose monophosphate pathway. Cytochromes of b-, c-, and o- types were found. Cell-free extracts contained a phenazine methosulfate-linked methanol dehydrogenase activity, which required ammonium ions as an activator. Cells harvested after the late exponential phase seemed to contain blue protein.ein.

• Journal of Microbiology and Biotechnology
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• v.33 no.4
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• pp.485-492
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• 2023

Methylorubrum extorquens, a facultative methylotroph, assimilates C₁ compounds and accumulates poly-β-hydroxylbutyrate (PHB) as carbon and energy sources. The ethylmalonyl pathway is central to the carbon metabolism of M. extorquens, and is linked with a serine cycle and a PHB biosynthesis pathway. Understanding the ethylmalonyl pathway is vital in utilizing methylotrophs to produce value-added chemicals. In this study, we determined the crystal structure of the mesaconyl-CoA hydratase from M. extorquens (MeMeaC) that catalyzes the reversible conversion of mesaconyl-CoA to β-methylmalyl-CoA. The crystal structure of MeMeaC revealed that the enzyme belongs to the MaoC-like dehydratase domain superfamily and functions as a trimer. In our current MeMeaC structure, malic acid occupied the substrate binding site, which reveals how MeMeaC recognizes the β-methylmalyl-moiety of its substrate. The active site of the enzyme was further speculated by comparing its structure with those of other MaoC-like hydratases.

• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1261-1271
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• 2020

Cytochrome c_L (Cytc_L) is an essential protein in the process of methanol oxidation in methylotrophs. It receives an electron from the pyrroloquinoline quinone (PQQ) cofactor of methanol dehydrogenase (MDH) to produce formaldehyde. The direct electron transfer mechanism between Cytc_L and MDH remains unknown due to the lack of structural information. To help gain a better understanding of the mechanism, we determined the first crystal structure of heme c containing Cytc_L from the aquatic methylotrophic bacterium Methylophaga aminisulfidivorans MP^T at 2.13 Å resolution. The crystal structure of Ma-Cytc_L revealed its unique features compared to those of the terrestrial homologues. Apart from Fe in heme, three additional metal ion binding sites for Na⁺, Ca⁺, and Fe²⁺ were found, wherein the ions mostly formed coordination bonds with the amino acid residues on the loop (G93-Y111) that interacts with heme. Therefore, these ions seemed to enhance the stability of heme insertion by increasing the loop's steadiness. The basic N-terminal end, together with helix α4 and loop (G126 to Y136), contributed positive charge to the region. In contrast, the acidic C-terminal end provided a negatively charged surface, yielding several electrostatic contact points with partner proteins for electron transfer. These exceptional features of Ma-Cytc_L, along with the structural information of MDH, led us to hypothesize the need for an adapter protein bridging MDH to Cytc_L within appropriate proximity for electron transfer. With this knowledge in mind, the methanol oxidation complex reconstitution in vitro could be utilized to produce metabolic intermediates at the industry level.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.140-146
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• 1999

When methanol was the sole carbon source, Methylobacterium organophilum NCIB 11278, a facultative methylotroph, accumulated Poly-$\beta$-hydroxybutyrate (PHB) as 59% (w/w) of dry cell weight under potassium limitation, 37% under sulfate limitation, and 33% under nitrogen limitation. Based on a stoichiometric analysis of PHB synthesis from methanol, it was suspected that PHB synthesis is accompanied by the overproduction of energy, either 6-10 ATP and 1 $FADH_2$ or 6 ATP and 3 NADPH to balance the NADH requirement, per PHB monomer. This was confirmed by observation of increased intracellular ATP levels during PHB accumulation. The intracellular ATP with limited potassium, sulfate, and ammonium increased to 0.185, 0.452, and 0.390 $\mu$moles ATP/g Xr (residual cell mass) during PHB accumulation, respectively. The intracellular ATP level under potassium limitation was similar to that when there was no nutrient limitation and no PHB accumulation, 0.152- 0.186 $\mu$moles ATP/g Xr. We propose that the maximum PHB accumulation observed when potassium was limited is a result of the energy balance during PHB accumulation. Microorganisms have high energy requirements under potassium limitation. Enhanced PHB accumulation, in ammonium and sulfate limited conditions with the addition of 2,4-dinitrophenol, which dissipates surplus energy, proves this assumption. With the addition of 1 mM of 2,4-dinitrophenol, the PHB content increased from 32.4% to 58.5% of dry cell weight when nitrogen limited and from 15.1 % to 31.0% of dry cell weight when sulfate limited.