Methylobacterium Organophilum Research Articles

The roles of Methylobacterium organophilum and Sphingomonas melonis for accelerating N-methyl pyrrolidone (NMP) biodegradation

Although N-methyl pyrrolidone (NMP) is biodegradable, normal activated sludge exhibits poor NMP biodegradation unless it is acclimated with NMP for an extended time. In this work, glucose-acclimated biomass (GAB) was acclimated with NMP for over two months to obtain NMP-acclimated biomass (NAB). NAB was superior to GAB for NMP biodegradation, and this correlated to higher abundances of the genera Mycobacterium, Amaricoccus, and Thauera in NAB. However, when GAB was bioaugmented with Methylobacterium organophilum or Sphingomonas melonis, which were isolated from NAB based on their growth on NAB, the bioaugmented GAB immediately showed rapid biodegradation of NMP. M. organophilum was superior to S. melonis for accelerating NMP biodegradation, and the acceleration of NMP biodegradation was proportional to the mass of M. organophilum or S. melonis added. A key finding was that the limiting step for NMP mineralization was biodegradation of monomethylamine, an NMP-biodegradation intermediate that also is inhibitory. Bioaugmenting M. organophilum or S. melonis accelerated NMP biodegradation by enhancing monomethylamine removal, with M. organophilum being superior to S. melonis. The documented rapid impact of bioaugmentation can make it especially valuable in response to an input of a toxicant.

Recent trends of biotechnological production of polyhydroxyalkanoates from C1 carbon sources.

Growing concerns over the use of limited fossil fuels and their negative impacts on the ecological niches have facilitated the exploration of alternative routes. The use of conventional plastic material also negatively impacts the environment. One such green alternative is polyhydroxyalkanoates, which are biodegradable, biocompatible, and environmentally friendly. Recently, researchers have focused on the utilization of waste gases particularly those belonging to C1 sources derived directly from industries and anthropogenic activities, such as carbon dioxide, methane, and methanol as the substrate for polyhydroxyalkanoates production. Consequently, several microorganisms have been exploited to utilize waste gases for their growth and biopolymer accumulation. Methylotrophs such as Methylobacterium organophilum produced highest amount of PHA up to 88% using CH4 as the sole carbon source and 52-56% with CH3OH. On the other hand Cupriavidus necator, produced 71-81% of PHA by utilizing CO and CO2 as a substrate. The present review shows the potential of waste gas valorization as a promising solution for the sustainable production of polyhydroxyalkanoates. Key bottlenecks towards the usage of gaseous substrates obstructing their realization on a large scale and the possible technological solutions were also highlighted. Several strategies for PHA production using C1 gases through fermentation and metabolic engineering approaches are discussed. Microbes such as autotrophs, acetogens, and methanotrophs can produce PHA from CO2, CO, and CH4. Therefore, this article presents a vision of C1 gas into bioplastics are prospective strategies with promising potential application, and aspects related to the sustainability of the system.

Differential Microbial Signature Associated With Benign Prostatic Hyperplasia and Prostate Cancer.

Apart from other risk factors, chronic inflammation is also associated with the onset of Prostate Cancer (PCa), wherein pathogen infection and tissue microbiome dysbiosis are known to play a major role in both inflammatory response and cancer development. However, except for a few studies, the link between microbes and PCa remained poorly understood. To explore the potential microbiome signature associated with PCa in Indian patients, we investigated differential compositions of commensal bacteria among patients with benign prostatic hyperplasia (BPH) and PCa using 16S rRNA amplicon sequencing followed by qPCR analyses using two distinct primer sets. Using two independent cohorts, we show that Prevotella copri, Cupriavidus campinensis, and Propionibacterium acnes represent the three most abundant bacteria in diseased prostate lesions. LEfSe analyses identified that while Cupriavidus taiwanensis and Methylobacterium organophilum are distinctly elevated in PCa samples, Kocuria palustris and Cellvibrio mixtus are significantly enriched in BPH samples. Furthermore, we identify that a number of human tumor viruses, including Epstein-Barr virus (EBV) and hepatitis B virus (HBV), along with two high-risk human papillomaviruses - HPV-16 and HPV-18, are significantly associated with the PCa development and strongly correlated with PCa bacterial signature. The study may thus offer to develop a framework for exploiting this microbial signature for early diagnosis and prognosis of PCa development.

Growth of Methylobacterium organophilum in Methanol for the Simultaneous Production of Single-Cell Protein and Metabolites of Interest.

This study aims to monitor the growth of the methylotrophic bacteria Methylobacterium organophilum in a culture medium with methanol as a carbon source and to verify the production of unicellular proteins and other biomolecules, such as carotenoids, exopolysaccharides and polyhydroxyalkanoates, making them more attractive as animal feed. Bacterial growth was studied in shake flasks using different carbon/nitrogen (C:N) ratios to determine their best ratio for achieving the highest volumetric productivity of cells and substrate consumption rate. This optimal parameter was further used in a fed-batch operating bioreactor system to define the kinetic profile of cell growth. Methanol consumption was measured by HPLC analysis and the extracted pigments were analyzed by liquid chromatography/mass spectrometry. Chemical composition and rheological properties of the produced exopolysaccharides were also determined. The best experimental parameters were verified using an initial methanol concentration of 7 g/L in the culture medium. The same initial substrate concentration was used in the fed-batch operation and after 60 h of cultivation 5 g/L of biomass were obtained. The accumulation of carotenoids associated with cell growth was monitored, reaching a concentration of 1.6 mg/L at the end of the process. These pigments were then analyzed and characterized as a set of xanthophylls (oxidized carotenoids). In addition, two other product types were identified during the fed-batch operation: exopolysaccharides, which reached a concentration of 8.9 g/L at the end of the cultivation, and an intracellular granular structure that was detected by transmission electron microscopy (TEM), suggesting the accumulation of polyhydroxyalkanoate (PHA), most likely polyhydroxybutyrate. Methylobacterium organophilum demonstrated a unique ability to produce compounds of commercial interest. The distinct metabolic diversity of this bacterium makes room for its use in biorefineries.

The role of hygrodynamic resistance compared to biofilm formation in helping pathogenic bacteria dominate air-conditioning units recovered from odour problems

ABSTRACT We previsouly found that installing filters in odourous air-conditioning units (ACUs) to block the entry of skin squames could well tackle the odour problems. In this study, we revisited and sampled the ACUs installed with filters earlier to study the bacterial communities inside the ACUs using 16S amplicon sequencing. We identified 26 genera and found that the skin bacteria isolated in the previous work were absent in this study. Two pathogenic bacteria, Methylobacterium and Sphingomonas, dominated ACUs instead. Afterwards, these two bacteria were identified to species level (Methylobacterium organophilum and Sphingomonas paucimobilis, respectively), and examined in terms of their biofilm formation ability and resistance to changing moisture conditions together with another prevalent species isolated in our previous study, namely Micrococcus luteus, in order to understand the mechanisms of the survival of bacteria in ACUs. In general, M. organophilum and M. luteus showed good biofilm formation ability at all tested temperature levels, but S. paucimobilis only displayed limited biofilm formation. Whereas, all these three bacteria well maintained their survival after wet-dry cycles. These results suggest that compared to biofilm formation, ability to survive under hygrodynamics tends to play a more important role in helping bacteria dominate ACUs. Further, this study implies that the absence of odour problem does not guarantee a healthy environment, more attentions should be given to limit the abundance of hydrodynamic-resistant pathogenic bacteria.

Comprehensive Comparative Genomics and Phenotyping of Methylobacterium Species.

The pink-pigmented facultative methylotrophs (PPFMs), a major bacterial group found in the plant phyllosphere, comprise two genera: Methylobacterium and Methylorubrum. They have been separated into three major clades: A, B (Methylorubrum), and C. Within these genera, however, some species lack either pigmentation or methylotrophy, which raises the question of what actually defines the PPFMs. The present study employed a comprehensive comparative genomics approach to reveal the phylogenetic relationship among the PPFMs and to explain the genotypic differences that confer their different phenotypes. We newly sequenced the genomes of 29 relevant-type strains to complete a dataset for almost all validly published species in the genera. Through comparative analysis, we revealed that methylotrophy, nitrate utilization, and anoxygenic photosynthesis are hallmarks differentiating the PPFMs from the other Methylobacteriaceae. The Methylobacterium species in clade A, including the type species Methylobacterium organophilum, were phylogenetically classified into six subclades, each possessing relatively high genomic homology and shared phenotypic characteristics. One of these subclades is phylogenetically close to Methylorubrum species; this finding led us to reunite the two genera into a single genus Methylobacterium. Clade C, meanwhile, is composed of phylogenetically distinct species that share relatively higher percent G+C content and larger genome sizes, including larger numbers of secondary metabolite clusters. Most species of clade C and some of clade A have the glutathione-dependent pathway for formaldehyde oxidation in addition to the H4MPT pathway. Some species cannot utilize methanol due to their lack of MxaF-type methanol dehydrogenase (MDH), but most harbor an XoxF-type MDH that enables growth on methanol in the presence of lanthanum. The genomes of PPFMs encode between two and seven (average 3.7) genes for pyrroloquinoline quinone-dependent alcohol dehydrogenases, and their phylogeny is distinctly correlated with their genomic phylogeny. All PPFMs were capable of synthesizing auxin and did not induce any immune response in rice cells. Other phenotypes including sugar utilization, antibiotic resistance, and antifungal activity correlated with their phylogenetic relationship. This study provides the first inclusive genotypic insight into the phylogeny and phenotypes of PPFMs.

Methylobacterium terricola sp. nov., a gamma radiation-resistant bacterium isolated from gamma ray-irradiated soil.

A gamma radiation-resistant and pink-pigmented bacterial strain, designated as 17Sr1-39T, was isolated from a gamma ray-irradiated soil sample collected in the Republic of Korea. Cells were Gram-stain-negative, strictly aerobic, flagellated, asporogenous, rod-shaped and methylotrophic. Results of 16S rRNA gene sequence analysis showed that strain 17Sr1-39T was phylogenetically related to Methylobacterium currus PR1016AT (97.3 %), Methylobacterium aquaticum DSM 16371T (97.2 %), Methylobacterium platani PMB02T (97.0 %), Methylobacterium frigidaeris IER25-16T (96.6 %), Methylobacterium terrae 17Sr1-28T (96.6 %) and Methylobacterium organophilum JCM 2833T (93.4 %). The G+C content calculated based on the genome sequence was 70.4 mol%. The average nucleotide identity and in silico DNA-DNA hybridization values between strain 17Sr1-39T and M. currus, M. aquaticum, M. platani, M. frigidaeris, M. terrae and M. organophilum were 77.3-89.9 and 22-38.2 %, respectively. The predominant fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c). The predominant quinone was ubiquinone 10 and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Based on the data from phenotypic tests and genotypic differences between strain 17Sr1-39T and its close phylogenetic relatives, strain 17Sr1-39T represented a new species belonging to the genus Methylobacterium, for which the name Methylobacterium terricola sp. nov. (=KACC 52905T=NBRC 112874T) is proposed.

Methylobacterium durans sp. nov., a radiation-resistant bacterium isolated from gamma ray-irradiated soil.

A gamma radiation-resistant, Gram-stain negative, oxidase and catalase positive, aerobic, flagellated, rod-shaped, methylotrophic and pink-pigmented bacterial strain designated 17SD2-17T was isolated from gamma-ray-irradiated soil collected in Korea. The 16S rRNA gene sequence analysis showed that strain 17SD2-17T is phylogenetically related to Methylobacterium organophilum DSM 760T (97.6%), Methylobacterium oxalidis 35aT (97.4%) and Methylobacterium soli YIM 48816T (97.0%). The G+C content calculated based on the draft genome sequence is 68.7mol%. The DNA-DNA hybridisation between 17SD2-17T and its close relatives was found to be less than 40%. The predominant fatty acid was identified as summed feature 8 (C18:1ω7c and/or C18:1ω6c) and the major respiratory quinone as Q-10. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. On the basis of the data from phenotypic tests and genotypic differences between strain 17SD2-17T and its close phylogenetic relatives, strain 17SD2-17T is concluded to represent a new species belonging to the genus Methylobacterium, for which the name Methylobacterium durans sp. nov. (= KCTC 52908T = NBRC 112876T) is proposed.

Methylobacterium terrae sp. nov., a radiation-resistant bacterium isolated from gamma ray-irradiated soil.

A Gram-stain-negative, asporogenous, aerobic rods, motile by means of a single polar flagellum, catalase- and oxidase-positive, methylotrophic bacterium, designated 17Sr1-28T, was isolated from gamma ray-irradiated soil. The 16S rRNA gene sequence analysis showed that strain 17Sr1-28T was phylogenetically related to Methylobacterium currus PR1016AT (96.8%), Methylobacterium platani PMB02T (96.2%), Methylobacterium aquaticum DSM 16371T (96.3%), Methylobacterium tarhaniae N4211T (96.4%), Methylobacterium frigidaeris IER25-16T (95.8%), and Methylobacterium organophilum JCM 2833T (92.7%). The G+C content calculated based on genome sequence was 71.6%. The average nucleotide identity and in silico DNA-DNA hybridization values between strain 17Sr1- 28T and M. currus, M. platani, M. aquaticum, M. tarhaniae, M. frigidaeris, and M. organophilum were 77.7-90.4% and 22-39.6%, respectively. The major fatty acids of strain 17Sr1-28T were summed feature 8 (C18:1ω7c and/or C18:1ω6c), and summed feature 3 (C16:1ω7c and/or C16:1ω6c). The predominant quinone was ubiquinone 10 and the major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylglycerol. On the basis of the data from phenotypic tests and genotypic differences between strain 17Sr1-28T and its close phylogenetic relatives, strain 17Sr1-28T represents a new species belonging to the genus Methylobacterium, for which the name Methylobacterium terrae sp. nov. (= KCTC 52904T = NBRC 112873T) is proposed.

Construction of methanol sensing Escherichia coli by the introduction of novel chimeric MxcQZ/OmpR two-component system from Methylobacterium organophilum XX

Methylobacterium organophilum XX is a type II facultative methylotroph that can grow on methanol. In M. organophilum XX, the MxcQ/MxcE two-component system (TCS) is involved in methanol metabolism. EnvZ/OmpR in E. coli TCS was exploited to develop a methanol biosensor by engaging the MxcQ/MxcE TCS system. The MxcQZ/OmpR methanol sensing chimeric TCS was constructed by integrating the sensing domain of M. organophilum MxcQ with the transmitter domain of E. coli EnvZ. The response regulator of the chimeric TCS system is OmpR, which regulates the expression of the ompC and gfp. The expression of ompC was monitored by real-time quantitative PCR analysis. The expression of gfp also confirmed the expression of the ompC. The maximum expression of ompC and gfp occurred with 0.05% of methanol, and the expression started to decline with further increases in methanol concentration. This system delivers rapid detection of methanol in the environment.

C35 Hopanoid Side Chain Biosynthesis: Reduction of Ribosylhopane into Bacteriohopanetetrol by a Cell-Free System Derived from Methylobacterium organophilum.

The major bacterial triterpenoids of the hopane series each consist of a C30 triterpene hopane moiety and an additional nonterpene C5 side chain derived from D-ribose and linked through its C-5 carbon atom to the hopane side chain. Bacteriohopanetetrol and aminobacteriohopanetriol are the most common representatives of this natural product series, adenosylhopane and ribosylhopane being putative precursors. Deuterium-labelled ribosylhopane was obtained by hemisynthesis and converted into deuterium-labelled bacteriohopanetetrol in the presence of NADPH, thus giving evidence of this as yet unknown precursor-to-product relationship in the bacterial hopanoid metabolic pathway.

Phylogenetic study of methanol oxidizers from chilika-lake sediments using genomic and metagenomic approaches.

Group-wise diversity of sediment methylotrophs of Chilika lake (Lat. 19°28'-19°54'N; Long. 85°06'-85°35'E) Odisha, India at various identified sites was studied. Both the culturable and unculturable (metagenome) methylotrophs were investigated in the lake sediments employing both mxaF and 16S rRNA genes as markers. ARDRA profiling, 16S rRNA gene sequencing, PAGE profiling of HaeIII, EcoRI restricted mxaF gene and the mxaF gene sequences using culture-dependent approach revealed the relatedness of α-proteobacteria and Methylobacterium, Hyphomicrobium and Ancyclobacter sp. The total viable counts of the culturable aerobic methylotrophs were relatively higher in sediments near the sea mouth (S3; Panaspada), also demonstrated relatively high salinity (0.1M NaCl) tolerance. Metagenomic DNA from the sediments, amplified using GC clamp mxaF primers and resolved through DGGE, revealed the diversity within the unculturable methylotrophic bacterium Methylobacterium organophilum, Ancyclobacter aquaticus, Burkholderiales and Hyphomicrobium sp. Culture-independent analyses revealed that up to 90% of the methylotrophs were unculturable. The study enhances the general understandings of the metagenomic methylotrophs from such a special ecological niche.

Hemisynthesis of deuteriated adenosylhopane and conversion into bacteriohopanetetrol by a cell-free system from Methylobacterium organophilum

Adenosylhopane is a putative precursor of the widespread bacterial C35 biohopanoids. A concise and flexible hemisynthesis of adenosylhopane has been developed including as key steps a cross metathesis between two olefins containing either the hopane moiety or a protected adenosine derivative and a subsequent diimide reduction of the resulting olefin. Reduction by deuteriated diimide allowed deuterium labelling. This synthetic protocol represents a versatile tool to access to deuteriated composite bacterial hopanoids required for biosynthetic studies. Deuteriated adenosylhopane was thus converted into bacteriohopanetetrol by a crude cell-free system from Methylobacterium organophilum in the presence of NADPH, showing for the first time the precursor to product relationship between these two bacterial metabolites.

Polyhydroxyalkanoates accumulation by Methylobacterium organophilum CZ-2 during methane degradation using citrate or propionate as cosubstrates

Methylobacterium organophilum CZ-2 synthesized polyhydroxyalkanoates (PHAs) under nitrogen limitation with CH4 as carbon source and when either citrate or propionate was added as cosubstrates. The highest PHAs content (yPHA) in closed flasks was obtained in the CH4-citrate and CH4-propionate experiments attaining values of 0.82 and 0.68, respectively. M. organophilum CZ-2 cultivated in bioreactors with citrate and continuous CH4 addition yielded a final PHAs concentration of 143gm−3 containing hydroxybutyrate (HB), hydroxyvalerate (HV) and hydroxyoctanoate (HO), in a 55:35:10 ratio, with, yPHA of 0.88 and a CH4 elimination capacity (EC) of 20gm−3h−1. With propionate, the yPHA was 0.3 and the EC around 8gm−3h−1. From 1H and 13C NMR experiments it was found that the polymer produced with CH4-citrate contained six different monomers: 3HB, 3HV, 4HV, 4-hydroxyheptanoate (4HH), 3HO and 4HO, showing the great versatility of this PHAs producing bacterium.

Production of poly-β-hydroxybutyrate (PHB) by Methylobacterium organophilum isolated from a methanotrophic consortium in a two-phase partition bioreactor

The biodegradation of methane, a greenhouse gas, and the accumulation of poly-β-hydroxybutyrate (PHB) were studied using a methanotrophic consortium and an isolated strain thereof. The specific rates for methane consumption were 100 and 17 m g C H 4 g x − 1 h − 1 for the isolate and the consortium, respectively. Also the effect of including 10% (v v −1) of silicone oil in a two-phase partitioning bioreactor (TPPB) was assayed for the elimination of 1% methane in air stream. TPPB allowed a 33–45% increase of methane elimination under growing conditions. Nitrogen limitation was assayed in bioreactors to promote PHB production. Under this condition, the specific methane degradation rate remained unchanged for the consortium and decreased to 36 m g C H 4 g x − 1 h − 1 for the isolated strain. The accumulated PHB in the reactor was 34% and 38% (w w −1) for the consortium and the isolate, respectively. The highest productivity was obtained in the TPPB and was 1 .61 m g PHB g x − 1 h − 1 . The CZ-2 isolate was identified as Methylobacterium organophilum, this is the first study that reports this species as being able to grow on methane and accumulate up to 57% (w w −1) of PHB under nitrogen limitation in microcosm experiments.

ChemInform Abstract: 2α-Methylhopanoids: First Recognition in the Bacterium Methylobacterium organophilum and Obtention via Sulfur Induced Isomerization of 2β-Methylhopanoids. An Account for Their Presence in Sediments.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

Methylobacterium jeotgali sp. nov., a non-pigmented, facultatively methylotrophic bacterium isolated from jeotgal, a traditional Korean fermented seafood

A novel facultatively methylotrophic, Gram-negative, rod-shaped bacterium, designated strain S2R03-9(T), was isolated from jeotgal, a traditional Korean fermented seafood. The organism was strictly aerobic, motile by means of a single polar flagellum, non-sporulating and catalase- and oxidase-positive. Strain S2R03-9(T) grew in the presence of 0-1 % (w/v) NaCl and at pH 6.0-10.0, with optimum growth in the absence of NaCl and at pH 7.0. It grew at temperatures in the range 20.0-30.0 degrees C, with optimum growth at 30 degrees C. Colonies grown on R2A medium were non-pigmented, opaque and creamy white. Phylogenetic analysis based on 16S rRNA gene sequences indicated that it was most closely related to Methylobacterium organophilum JCM 2833(T) (96.6 % similarity) and the phylogenetic similarities to all other Methylobacterium species with validly published names were less than 95.0 %. The DNA G+C content was 64.9 mol%. The phylogenetic analysis, the phenotypic assessment and the chemotaxonomic data (major ubiquinone, Q-10; major fatty acids, C(18 : 1) and C(18 : 0)) showed that S2R03-9(T) represents a novel species within the genus Methylobacterium in the class Alphaproteobacteria, for which the name Methylobacterium jeotgali sp. nov. is proposed. The type strain is S2R03-9(T) (=KCTC 12671(T)=LMG 23639(T)).

Facultative Methanotrophs Revisited

Facultative Methanotrophs Revisited

Methylobacterium populi sp. nov., a novel aerobic, pink-pigmented, facultatively methylotrophic, methane-utilizing bacterium isolated from poplar trees (Populus deltoides x nigra DN34).

A pink-pigmented, aerobic, facultatively methylotrophic bacterium, strain BJ001T, was isolated from internal poplar tissues (Populus deltoidesxnigra DN34) and identified as a member of the genus Methylobacterium. Phylogenetic analyses showed that strain BJ001T is related to Methylobacterium thiocyanatum, Methylobacterium extorquens, Methylobacterium zatmanii and Methylobacterium rhodesianum. However, strain BJ001T differed from these species in its carbon-source utilization pattern, particularly its use of methane as the sole source of carbon and energy, an ability that is shared with only one other member of the genus, Methylobacterium organophilum. In addition, strain BJ001T is the only member of the genus Methylobacterium to be described as an endophyte of poplar trees. On the basis of its physiological, genotypic and ecological properties, the isolate is proposed as a member of a novel species of the genus Methylobacterium, Methylobacterium populi sp. nov. (type strain, BJ001T=ATCC BAA-705T=NCIMB 13946T).

Isolation and 16S DNA Characterization of Soil Microorganisms from Tropical Soils Capable of Utilizing the Herbicides Hexazinone and Tebuthiuron

Six non‐fermentative bacteria were isolated from Colombian (South America) and Hawaiian (USA) soils after enrichment with minimal medium supplemented with two herbicides, hexazinone (Hex) and tebuthiuron (Teb). Microscopic examination and physiological tests were followed by partial 16S DNA sequence analysis, using the first 527 bp of the 16S rRNA gene for bacterial identification. The isolated microorganisms (and in brackets, the herbicide that each degraded) were identified as: from Colombia, Methylobacterium organophilum [Teb], Paenibacillus pabuli [Teb], and Microbacterium foliorum [Hex]; and from Hawaii, Methylobacterium radiotolerans [Teb], Paenibacillus illinoisensis [Hex], and Rhodococcus equi [Hex]. The findings further explain how these herbicides, which have potential for illicit coca (Erythroxylum sp.) control, dissipate following their application to tropical soils.