Methane hydroxylation by Methylosinus trichosporium OB3b: Monitoring the biocatalyst activity for methanol production optimization in an innovative membrane bioreactor (original) (raw)
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Optimization of lab scale methanol production by Methylosinus trichosporium OB3b
Biotechnology and Bioprocess Engineering, 2010
Methylosinus trichosporium OB3b is a methanotrophic bacterium containing particulate methane monooxygenase (MMO), which catalyzes the hydroxylation of methane to methanol. The methanol is further oxidized to formaldehyde by methanol dehydrogenase (MDH). We developed a novel compulsory circulation diffusion system for cell cultivation. A methane/air mixture (1:1, v/v) was prepared in a tightly sealed gas reservoir and pumped into a nitrate mineral salt culture medium under optimal conditions (5 μM CuSO4, pH 7.0, 30°C). Cells were harvested, washed, and resuspended (0.6 mg dry cells/mL) in a 500 mL flask in 100 mL of 10 mM phosphate buffer (pH 7.0) containing 100 mM NaCl and 1 mM EDTA as MDH inhibitors, and 20 mM sodium formate. A single 12 h batch reaction at 25°C yielded a final concentration of 13.2 mM methanol. The use of a repeated batch mode, in which the accumulated methanol was removed after each of three 8 h cycles over a 24 h period, showed a productivity of 2.17 μmol methanol/h/mg dry cell wt. Finally, a lab-scale reaction performed using a 3 L cylindrical reactor with a working volume of 1 L produced 13.7 mM methanol after 16 h. Our results identify a simple process for improving the productivity of biologically derived methanol and, therefore the utility of methane as an energy source.
Biosynthesis of methanol from methane by Methylosinus trichosporium OB3b
Chemical Papers, 2009
Methanol has recently attracted significant interest in the energetic field. Current technology for the conversion of methane to methanol is based on energy intensive endothermic steam reforming followed by catalytic conversion into methanol. The one-step method performed at very low temperatures (35°C) is methane oxidation to methanol via bacteria. The aim of this work was to examine the role of copper in the one-step methane oxidation to methanol by utilizing whole cells of Methylosinus trichosporium OB3b bacteria. From the results obtained it was found that copper concentration in the medium influences the rate of bacterial biomass growth or methanol production during the process of methane oxidation to methanol. The presented results indicate that the process of methane oxidation to methanol by Methylosinus trichosporium OB3b bacteria is most efficient when the mineral medium contains 1.0 × 10−6 mol dm−3 of copper. Under these conditions, a satisfactory growth of biomass was also achieved.
Immobilization of Methylosinus trichosporium OB3b for methanol production
Journal of industrial microbiology & biotechnology, 2018
Due to the natural gas boom in North America, there is renewed interest in the production of other chemical products from methane. We investigated the feasibility of immobilizing the obligate methanotrophic bacterium Methylosinus trichosporium OB3b in alginate beads, and selectively inactivating methanol dehydrogenase (MDH) with cyclopropane to produce methanol. In batch cultures and in semi-continuous flow columns, the exposure of alginate-immobilized cells to cyclopropane or cyclopropanol resulted in the loss of the majority of MDH activity (> 80%), allowing methanol to accumulate to significant concentrations while retaining all of M. trichosporium OB3b's methane monooxygenase capacity. Thereafter, the efficiency of methanol production fell due to recovery of most of the MDH activity; however, subsequent inhibition periods resulted in renewed methanol production efficiency, and immobilized cells retained methane-oxidizing activity for at least 14 days.
Enrichment of anaerobic methanotrophs in sulfate-reducing membrane bioreactors
Biotechnology and Bioengineering, 2009
Anaerobic oxidation of methane (AOM) in marine sediments is coupled to sulfate reduction (SR). AOM is mediated by distinct groups of archaea, called anaerobic methanotrophs (ANME). ANME co-exist with sulfate-reducing bacteria, which are also involved in AOM coupled SR. The microorganisms involved in AOM coupled to SR are extremely difficult to grow in vitro. Here, a novel well-mixed submerged-membrane bioreactor system is used to grow and enrich the microorganisms mediating AOM coupled to SR. Four reactors were inoculated with sediment sampled in the Eckernförde Bay (Baltic Sea) and operated at a methane and sulfate loading rate of 4.8 L L−1 day−1 (196 mmol L−1 day−1) and 3.0 mmol L−1 day−1. Two bioreactors were controlled at 15°C and two at 30°C, one reactor at 30°C contained also anaerobic granular sludge. At 15°C, the volumetric AOM and SR rates doubled approximately every 3.8 months. After 884 days, an enrichment culture was obtained with an AOM and SR rate of 1.0 mmol g day−1 (286 µmol g day−1). No increase in AOM and SR was observed in the two bioreactors operated at 30°C. The microbial community of one of the 15°C reactors was analyzed. ANME-2a became the dominant archaea. This study showed that sulfate reduction with methane as electron donor is possible in well-mixed bioreactors and that the submerged-membrane bioreactor system is an excellent system to enrich slow-growing microorganisms, like methanotrophic archaea. Biotechnol. Bioeng. 2009; 104: 458–470 © 2009 Wiley Periodicals, Inc.
Production of Methanol from Methane by Encapsulated Methylosinus sporium
Journal of microbiology and biotechnology, 2016
Massive reserves of methane (CH₄) remain unexplored as a feedstock for the production of liquid fuels and chemicals, majorly because of the lack of economically suitable and sustainable strategies for selective oxidation of CH₄ to methanol. The present study demonstrates the bioconversion of CH₄ to methanol mediated by Type I methanotrophs, such as Methylomicrobium album and Methylomicrobium alcaliphilum. Furthermore, immobilization of 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°C, and 175 rpm using inoculum (1.5 mg of dry cell mass/ml) and 20% of CH₄ as a feed, respectively. Under these conditions, maximum methanol production (3.43 and 3.73 mM) by the encapsulated cells was recorded. Even after six cycles of reuse, t...
Bioresource Technology, 2017
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Methanogenesis mediated by methylotrophic mixed culture
Applied Biochemistry and Biotechnology, 1994
Enrichment of methanogenic cultures on methanol from the microbial population in the anaerobic digesters operated on agricultural wastes revealed a high rate of biomethanation efficiency. Routine maintenance of this enrichment in a minimal basal medium at room temperature resulted in maximal growth in 40-50 d, and indicated pigment production toward the end of the growth phase. The cultures grown in three different media, with different substrates under light and dark conditions, were analyzed for protein, pigment, and gaseous products, and morphological studies were carried out by light, phase-contrast, fluorescence, and electron microscopy. In different media with methanol as substrate, growth and pigment production were maximal for the light-grown cells, decreasing in the order: phototrophic (PS(m)) > mineral > basal medium. Methanation and phototrophic growth were inversely correlated under lightgrown conditions. In contrast, growth in the dark was predominently methanogenic in the decreasing order: mineral > basal > PS (m). Among other growth conditions tested, utilization of phototrophic substrates under light and dark conditions indicated the following: 1. Basal and mineral media were supportive of methanogenic growth under both light and dark conditions, although methane yields under light-grown conditions were low; *Author to whom all correspondence and reprint requests should be addressed.