Methylotroph

The key intermediate in methylotrophic metabolism is formaldehyde, which can be diverted to either assimilatory or dissimilatory pathways. Methylotrophs produce formaldehyde through oxidation of methanol and/or methane. Methane oxidation requires the enzyme methane monooxygenase (MMO). Methylotrophs with this enzyme are given the name methanotrophs. The oxidation of methane (or methanol) can be assimilatory or dissimilatory in nature (see figure). If dissimilatory, the formaldehyde intermediate is oxidized completely into CO2 to produce reductant and energy. If assimilatory, the formaldehyde intermediate is used to synthesize a 3-Carbon (C3) compound for the production of biomass. Many methylotrophs use multi-carbon compounds for anabolism, thus limiting their use of formaldehyde to dissimilatory processes, however methanotrophs are generally limited to only C1metabolism. or a non-specific alcohol oxidase in yeast. Electrons from methanol oxidation are passed to a membrane-associated quinone of the electron transport chain to produce ATP. In dissimilatory processes, formaldehyde is completely oxidized to CO2 and excreted. Formaldehyde is oxidized to formate via the action of Formaldehyde dehydrogenase (FALDH), which provides electrons directly to a membrane associated quinone of the electron transport chain, usually cytochrome b or c. and CO2. Anabolism The main metabolic challenge for methylotrophs is the assimilation of single carbon units into biomass. Through de novo synthesis, methylotrophs must form carbon-carbon bonds between 1-Carbon (C1) molecules. This is an energy intensive process, which facultative methylotrophs avoid by using a range of larger organic compounds. However, obligate methylotrophs must assimilate C1 molecules. All four pathways incorporate 3 C1 molecules into multi-carbon intermediates, then cleave one intermediate into a new C3 molecule. The remaining intermediates are rearranged to regenerate the original multi-carbon intermediates. Bacteria Each species of methylotrophic bacteria has a single dominant assimilation pathway. Ribulose bisphosphate (RuBP) cycle Unlike the other assimilatory pathways, bacteria using the RuBP pathway derive all of their organic carbon from CO2 assimilation. This pathway was first elucidated in photosynthetic autotrophs and is better known as the Calvin Cycle. Shortly thereafter, methylotrophic bacteria who could grow on reduced C1 compounds were found using this pathway. First, 3 molecules of ribulose 5-phosphate are phosphorylated to ribulose 1,5-bisphosphate (RuBP). The enzyme ribulose bisphosphate carboxylase (RuBisCO) carboxylates these RuBP molecules which produces 6 molecules of 3-phosphoglycerate (PGA). The enzyme phosphoglycerate kinase phosphorylates PGA into 1,3-diphosphoglycerate (DPGA). Reduction of 6 DPGA by the enzyme glyceraldehyde phosphate dehydrogenase generates 6 molecules of the C3 compound glyceraldehyde-3-phosphate (GAP). One GAP molecule is diverted towards biomass while the other 5 molecules regenerate the 3 molecules of ribulose 5-phosphate. Through radio-labelling experiments, it was shown that M. methanica used the ribulose monophosphate (RuMP) pathway. This has led researchers to propose that the RuMP cycle may have preceded the RuBP cycle. First, 2 molecules of formaldehyde are added to 2 molecules of the amino acid glycine. This produces two molecules of the amino acid serine, the key intermediate of this pathway. These serine molecules eventually produce 2 molecules of 2-phosphoglycerate, with one C3 molecule going towards biomass and the other being used to regenerate glycine. Notably, the regeneration of glycine requires a molecule of CO2 as well, therefore the Serine pathway also differs from the other 3 pathways by its requirement of both formaldehyde and CO2. Compartmentalization of this reaction in peroxisomes likely sequesters the hydrogen peroxide produced. Catalase is produced in the peroxisomes to deal with this harmful by-product. This pathway assimilates three molecules of formaldehyde into 1 molecule of DHAP using 3 molecules of xylulose 5-phosphate as the key intermediate. DHA synthase acts as a transferase (transketolase) to transfer part of xylulose 5-phosphate to DHA. Then these 3 molecules of DHA are phosphorylated to DHAP by triokinase. Like the other cycles, 3 C3 molecules are produced with 1 molecule being directed for use as cell material. The other 2 molecules are used to regenerate xylulose 5-phosphate. ==Environmental Implications==

As key players in the carbon cycle, methylotrophs work to reduce global warming primarily through the uptake of methane and other greenhouse gases. In aqueous environments, methanogenic archaea produce 40-50% of the world's methane. Symbiosis between methanogens and methanotrophic bacteria greatly decreases the amount of methane released into the atmosphere. This symbiosis is also important in the marine environment. Marine bacteria are very important to food webs and biogeochemical cycles, particularly in coastal surface waters but also in other key ecosystems such as hydrothermal vents. There is evidence of widespread and diverse groups of methylotrophs in the ocean that have potential to significantly impact marine and estuarine ecosystems. One-carbon compounds used as a carbon and energy source by methylotrophs are found throughout the ocean. These compounds include methane, methanol, methylated amines, methyl halides, and methylated sulfur compounds, such as dimethylsulfide (DMS) and dimethylsulfoxide (DMSO). Some of these compounds are produced by phytoplankton and some come from the atmosphere. Studies incorporating a wider range of one-carbon substrates have found increasing diversity of methylotrophs, suggesting that the diversity of this bacterial group has not yet fully been explored. while in others a product of methylotroph activity, methylamine, has been found to be emitted from the ocean and form aerosols. This means that methylotrophic metabolism is expected to follow similar dynamics, which will then impact biogeochemical cycles and carbon fluxes. These types of studies will contribute to further understanding of deep sea carbon cycling and the connectivity between deep ocean and surface carbon cycling. The expansion of omics technologies has accelerated research on the diversity of methylotrophs, their abundance and activity in a variety of environmental niches, and their interspecies interactions. Further research must be done on these bacteria and the overall effect of bacterial drawdown and transformation of one-carbon compounds in the ocean. Current evidence points to a potentially substantial role for methylotrophs in the ocean in the cycling of carbon but also potentially in the global nitrogen, sulfur and phosphorus cycles as well as the air-sea flux of carbon compounds, which could have global climate impacts. Methylotrophs have high potential as alternative biofertilizers and bioinoculants due to their ability to form mutualistic relationships with several plant species. Methylotrophs provide plants with nutrients such as soluble phosphorus and fixed nitrogen and also play a role in the uptake of said nutrients. Additionally, they can help plants respond to environmental stressors through the production of phytohormones. Methylotrophic growth also inhibits the growth of harmful plant pathogens and induces systemic resistance. Methylotrophic biofertilizers used either alone or together with chemical fertilizers have been shown to increase both crop yield and quality without loss of nutrients. ==References==