Recent Advances in the Genetic Manipulation of Methylosinus trichosporium OB3b

Ro, Soo Y.; Rosenzweig, Amy C.

doi:10.1016/bs.mie.2018.02.011

Cited by 21 publications

(9 citation statements)

References 41 publications

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“…The Methylosinus trichosporium OB3b Δ pmoD mutant was generated by chromosomal gene disruption into wild-type Methylosinus trichosporium OB3b (a gift from John Lipscomb) using methods described previously 41 . Briefly, a gentamicin resistance gene ( gen R ) was inserted in the middle of PmoD MettrDRAFT_0381 via homologous recombination.…”

Section: Methodsmentioning

confidence: 99%

“…The gentamicin resistance cassette was amplified from vector pFBOH-LIC using primers listed in Supplementary Table 5 . The PCR products were assembled into pK18mobsacB_p15a 41 using Gibson assembly with primers listed in Supplementary Table 5 and transformed into Escherichia coli S17-1 ATCC 47055 to produce plasmid pSYR16. This plasmid was then introduced into Methylosinus trichosporium OB3b via conjugation with E. coli S17-1 cells transformed with pSYR16 on NMS mating agar plates (0 μM CuSO 4 ).…”

Section: Methodsmentioning

confidence: 99%

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Characterization of a long overlooked copper protein from methane- and ammonia-oxidizing bacteria

et al. 2018

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Methane-oxidizing microbes catalyze the oxidation of the greenhouse gas methane using the copper-dependent enzyme particulate methane monooxygenase (pMMO). Isolated pMMO exhibits lower activity than whole cells, however, suggesting that additional components may be required. A pMMO homolog, ammonia monooxygenase (AMO), converts ammonia to hydroxylamine in ammonia-oxidizing bacteria (AOB) which produce another potent greenhouse gas, nitrous oxide. Here we show that PmoD, a protein encoded within many pmo operons that is homologous to the AmoD proteins encoded within AOB amo operons, forms a copper center that exhibits the features of a well-defined CuA site using a previously unobserved ligand set derived from a cupredoxin homodimer. PmoD is critical for copper-dependent growth on methane, and genetic analyses strongly support a role directly related to pMMO and AMO. These findings identify a copper-binding protein that may represent a missing link in the function of enzymes critical to the global carbon and nitrogen cycles.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Characterization of a long overlooked copper protein from methane- and ammonia-oxidizing bacteria

et al. 2018

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“…Electroporation-based genetic manipulation methods have been developed in both type I and type II methanotrophs [9,31]. Recently, one research group from the National Renewable Energy Laboratory has successfully developed a dual-plasmid, broad-host-range CRISPR/Cas9 gene-editing system for Methylococcus capsulatus (Bath) [32].…”

Section: Genetic Tool Development For Methanotrophsmentioning

confidence: 99%

Bioproduction of Isoprenoids and Other Secondary Metabolites Using Methanotrophic Bacteria as an Alternative Microbial Cell Factory Option: Current Stage and Future Aspects

2019

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Methane is a promising carbon feedstock for industrial biomanufacturing because of its low price and high abundance. Recent advances in metabolic engineering and systems biology in methanotrophs have made it possible to produce a variety of value-added compounds from methane, including secondary metabolites. Isoprenoids are one of the largest family of secondary metabolites and have many useful industrial applications. In this review, we highlight the current efforts invested to methanotrophs for the production of isoprenoids and other secondary metabolites, including riboflavin and ectoine. The future outlook for improving secondary metabolites production (especially of isoprenoids) using metabolic engineering of methanotrophs is also discussed. gas-to-liquid bioconversion using methanotrophs, such as limited mass transfer rates of methane, low volumetric productivity, and instability of recombinant strains [11].Secondary metabolites produced by plants and microorganisms have many useful biological activities. Among the many secondary metabolites, isoprenoids are the largest family of natural products, with more than 50,000 members identified [12][13][14]. Many isoprenoids have been used in many applications such as pharmaceuticals, nutraceuticals, fragrances, as well as the chemical industry. Additionally, due to the methyl-branched and cyclized hydrocarbon alkene structure, some isoprenoids have attracted more attention for their potential use as advanced biofuels. Plants are the major sources of isoprenoids. However, there are still many limitations for the production of high quantities of natural isoprenoids, including slow growth and tissue-specific biosynthesis, and difficulties in harvesting and extraction [12][13][14]. In contrast, a promising alternative method for isoprenoid production is the reconstruction of isoprenoid biosynthesis pathways from plants into microbes. The application of this approach has been extensively studied on model organisms like Escherichia coli or Saccharomyces cerevisiae for a variety of isoprenoids [12,14]. E. coli and S. cerevisiae are promising hosts since they possess simple genetic backgrounds, high growth rates as well as well-developed genetic tools. For example, high production of isoprenoid-based biofuels has been achieved in the metabolic engineering of microbes via endogenous or heterologous biosynthetic pathways in these hosts. However, the vast majority of these microbe-based isoprenoid production cases rely on sugar-based metabolism, which is likely to increase in the price of isoprenoids produced in large quantity.The use of alternative carbon sources is attractive in industrial biotechnology for isoprenoid production. Furthermore, different type of pathway regulation in different microbes utilizing unusual carbon substrate like methane can play a vital role in metabolic engineering of various microbes including model organisms. Thus, metabolic engineering of methane-utilizing methanotrophs has recently attracted much attention, due to not only cheaper price of...

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“…M. trichosporium was considered the best methanotroph due to its ability to express both the soluble MMO (sMMO) enzyme and particulate MMO (pMMO). Additionally, this bacterium can express two distinct MDHs 15 . Although M. trichosporium was proven to be able to convert methanol from methane, process optimization is still crucial in order to achieve an optimum rate in methanol production.…”

Section: Introductionmentioning

confidence: 99%

Improving methanol production by Methylosinus trichosporium through the one factor at a time (OFAT) approach

et al. 2022

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The technology of methanol production from the fermentation of methanol‐producing bacteria (methanotroph) is still in the research phase. Maximizing the methanol production through bacteria fermentation in this process is a challenge to researchers. By optimizing the biological process, this technology can potentially fill the demand gap in methanol industries. Methylosinus trichosporium is a methanotroph that is used for the production of methanol in this analysis. Using OD600 analysis, the rate and growth profile for M. trichosporium is determined. The one factor at a time (OFAT) method was chosen to optimize the methanol production (percentage of methane, pH, and phosphate ion), and the optimum condition for this bacterium was determined. M. trichosporium showed a specific growth rate at 0.02 h–1 with the exponential phase reaching the maximum at 90 h of the cultivation period. Optimization analysis showed that the maximum methanol production by this bacterium was at a 1:1 methane–air ratio, a pH of 6.3–6.5, and 40 mM of phosphate ions. The ability of M. trichosporium to produce methanol was proved in this analysis with the maximum rate of production at 0.072 mg L–1 h–1. © 2022 Society of Chemical Industry and John Wiley & Sons, Ltd.

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Recent Advances in the Genetic Manipulation of Methylosinus trichosporium OB3b

Cited by 21 publications

References 41 publications

Characterization of a long overlooked copper protein from methane- and ammonia-oxidizing bacteria

Characterization of a long overlooked copper protein from methane- and ammonia-oxidizing bacteria

Bioproduction of Isoprenoids and Other Secondary Metabolites Using Methanotrophic Bacteria as an Alternative Microbial Cell Factory Option: Current Stage and Future Aspects

Improving methanol production by Methylosinus trichosporium through the one factor at a time (OFAT) approach

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