• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2098-2105
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• 2016

Massive reserves of methane ($CH_4$) remain unexplored as a feedstock for the production of liquid fuels and chemicals, mainly because of the lack of economically suitable and sustainable strategies for selective oxidation of $CH_4$ to methanol. The present study demonstrates the bioconversion of $CH_4$ to methanol mediated by Type I methanotrophs, such as Methylomicrobium album and Methylomicrobium alcaliphilum. Furthermore, immobilization of a Type II methanotroph, Methylosinus sporium, was carried out using different encapsulation methods, employing sodium-alginate (Na-alginate) and silica gel. The encapsulated cells demonstrated higher stability for methanol production. The optimal pH, temperature, and agitation rate were determined to be pH 7.0, $30^{\circ}C$, and 175 rpm, respectively, using inoculum (1.5 mg of dry cell mass/ml) and 20% of $CH_4$ as a feed. Under these conditions, maximum methanol production (3.43 and 3.73 mM) by the encapsulated cells was recorded. Even after six cycles of reuse, the Na-alginate and silica gel encapsulated cells retained 61.8% and 51.6% of their initial efficiency for methanol production, respectively, in comparison with the efficiency of 11.5% observed in the case of free cells. These results suggest that encapsulation of methanotrophs is a promising approach to improve the stability of methanol production.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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• pp.287-293
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• 2022

The hydroxylation of methane (CH₄) is crucial to the field of environmental microbiology, owing to the heat capacity of methane, which is much higher than that of carbon dioxide (CO₂). Soluble methane monooxygenase (sMMO), a member of the bacterial multicomponent monooxygenase (BMM) superfamily, is essential for the hydroxylation of specific substrates, including hydroxylase (MMOH), regulatory component (MMOB), and reductase (MMOR). The diiron active site positioned in the MMOH α-subunit is reduced through the interaction of MMOR in the catalytic cycle. The electron transfer pathway, however, is not yet fully understood due to the absence of complex structures with reductases. A type II methanotroph, Methylosinus sporium 5, successfully expressed sMMO and hydroxylase, which were purified for the study of the mechanisms. Studies on the MMOH-MMOB interaction have demonstrated that Tyr76 and Trp78 induce hydrophobic interactions through π-π stacking. Structural analysis and sequencing of the ferredoxin domain in MMOR (MMOR-Fd) suggested that Tyr93 and Tyr95 could be key residues for electron transfer. Mutational studies of these residues have shown that the concentrations of flavin adenine dinucleotide (FAD) and iron ions are changed. The measurements of dissociation constants (K_ds) between hydroxylase and mutated reductases confirmed that the binding affinities were not significantly changed, although the specific enzyme activities were significantly reduced by MMOR-Y93A. This result shows that Tyr93 could be a crucial residue for the electron transfer route at the interface between hydroxylase and reductase.

• 대한환경공학회지
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• 제33권9호
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• pp.662-669
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• 2011

본 연구에서는 메탄의 생물학적 메탄올 전환에 관한 연구를 수행하였다. 바이오가스 중의 메탄은 메탄산화균의 methane monooxygenase (MMO)의 생물학적 촉매반응에 의해 산화되었으며, 인산염, NaCl, $NH_4Cl$, EDTA와 같은 methanol dehydrogenase (MDH)의 활성 저해제를 이용하여 MDH의 활성도를 저해함으로써 메탄올의 전환이 이루어졌다. 메탄산화균은 $35^{\circ}C$, pH 7, 인공 바이오가스($CH_4$ 50%, $CO_2$ 50%) / Air의 부피비가 0.4인 조건에서 메탄 산화 정도가 0.56 mmol로 최대로 나타났다. 인산염 40 mM, NaCl 50 mM, $NH_4Cl$ 40 mM, EDTA $150{\mu}m$ 이하일 때 저해제의 종류에 상관없이 메탄 산화율은 80% 이상을 달성하였다. 한편, 인산염 40 mM, NaCl 100 mM, $NH_4Cl$ 40 mM, EDTA $50{\mu}m$ 주입 시 각각 1.30, 0.67, 0.74, 1.30 mmol의 메탄이 산화되는 동시에 각각 0.71, 0.60, 0.66, 0.66 mmol의 메탄올이 최대로 생성되었다. 이때의 메탄올 전환율은 각각 54.7, 89.9, 89.6 및 47.8%였으며 최대 메탄올 생성 속도는 $7.4{\mu}mol/mg{\cdot}h$였다. 이로부터 대상 저해제로 MDH 활성도를 일반적으로 35% 저해 시에 메탄올 생산량이 최대인 89.9%까지 나타남을 알 수 있었다.