Search Results (7)

The use of methane as a carbon source for producing bacterial single-cell protein (SCP) has been one of the most interesting developments in recent years. Most of these upcoming industries are using a methanotroph, Methylococcus capsulatus Bath, for SCP production using natural gas as the substrate. In the present study, we have explored the possibility of using an indigenously isolated methanotroph from a rice field in India, Methylomagnum ishizawai strain KRF4, for producing SCP from biogas [derived from cow dung]. The process was eco-friendly, required minimal instruments and chemicals, and was carried out under semi-sterile conditions in a tabletop fish tank. As the name suggests, Methylomagnum is a genus of large methanotrophs, and the strain KRF4 had elliptical to rectangular size and dimensions of ~4–5 µm × 1–2 µm. In static cultures, when biogas and air were supplied in the upper part of the growing tank, the culture grew as a thick pellicle/biofilm that could be easily scooped. The grown culture was mostly pure, from the microscopic observations where the large size of the cells, with rectangular-shaped cells and dark granules, could easily help identify any smaller contaminants. Additionally, the large cell size could be advantageous for separating biomass during downstream processing. The amino acid composition of the lyophilized biomass was analyzed using HPLC, and it was seen that the amino acid composition was comparable to commercial fish meal, soymeal, Pruteen, and the methanotroph-derived SCP-UniProtein^®. The only difference was that a slightly lower percentage of lysine, tryptophan, and methionine was observed in Methylomagnum-derived SCP. Methylomagnum ishizawai could be looked at as an alternative for SCP derived from methane or biogas due to the comparable SCP produced, on the qualitative level. Further intensive research is needed to develop a continuous, sustainable, and economical process to maximize biomass production and downstream processing. Full article

Thirty-two healthy adult dogs (16 males and 16 females) were fed control kibble diets for one month, followed by six months (Weeks 0 to 25) of diets containing either 0, 4, 6, or 8% cultured protein derived from Methylococcus capsulatus (FeedKind Pet^®, FK), then they were fed control diets (0% FK) for a further two months (Weeks 25 to 34). The diets were isonitrogenous, isolipidic, and isocaloric and stage- and age-specific. The dogs were assessed for overall health, weight gain, and body condition score (BCS). Blood samples were collected 1 week prior to randomization, during acclimation, then in Weeks 5, 13, 25, 30, 32, and 34 for hematology, coagulation, and clinical chemistry; urine was collected according to the same time schedule for urinalysis. Feces were assessed for parasite load and presence of occult blood during Weeks 5, 9, 13, 17, 21, and 25. Fecal samples were collected during acclimation and Weeks 25 and 34 for fecal microbiome analysis and in Week 25 for apparent total gastrointestinal tract digestibility (ATTD). All dogs maintained a healthy weight and BCS throughout the study. Hematology parameters were within normal limits at the end of each phase of the study. With the exception of a decrease in serum phosphorus level and in urine pH in all groups at the end of the study, urine and serum chemistry results were within normal limits at the end of each phase. ATTD values for organic matter, protein, and energy exceeded 80%, whilst digestibility values for copper were around 20%. The fecal microbiome was dominated by Firmicutes. Alpha diversity increased during the safety phase before returning to baseline levels during the washout phase. The dominant genera in all groups were Megamonas, Peptoclostridium, Turicibacter, Catenibacterium, Fusobacterium, Romboutsia, and Blautia. The study has shown that the inclusion of cultured protein at up to 8% of the total diet of adult dogs can provide sufficient nutrition and is safe with no long-term effects on a range of health parameters. Full article

Biotechnology continues to drive innovation in the production of pharmaceuticals, biofuels, and other valuable compounds, leveraging the power of microbial systems for enhanced yield and sustainability. Genome-scale metabolic (GSM) modeling has become an essential approach in this field, which enables a guide for targeting genetic modifications and the optimization of metabolic pathways for various industrial applications. While single-species GSM models have traditionally been employed to optimize strains like Escherichia coli and Lactococcus lactis, the integration of these models into community-based approaches is gaining momentum. Herein, we present a pipeline for community metabolic modeling with a user-friendly GUI, applying it to analyze interactions between Methylococcus capsulatus, a biotechnologically important methanotroph, and Escherichia coli W3110 under oxygen- and nitrogen-limited conditions. We constructed models with unmodified and homoserine-producing E. coli strains using the pipeline implemented in the original BioUML platform. The E. coli strain primarily utilized acetate from M. capsulatus under oxygen limitation. However, homoserine produced by E. coli significantly reduced acetate secretion and the community growth rate. This homoserine was taken up by M. capsulatus, converted to threonine, and further exchanged as amino acids. In nitrogen-limited modeling conditions, nitrate and ammonium exchanges supported the nitrogen needs, while carbon metabolism shifted to fumarate and malate, enhancing E. coli TCA cycle activity in both cases, with and without modifications. The presence of homoserine altered cross-feeding dynamics, boosting amino acid exchanges and increasing pyruvate availability for M. capsulatus. These findings suggest that homoserine production by E. coli optimizes resource use and has potential for enhancing microbial consortia productivity. Full article

This study was conducted to evaluate the effects of replacing fish meal (FM) with methanotroph (Methylococcus capsulatus, Bath) bacteria protein (MBP) in the diets of the juvenile American eel (Anguilla rostrata). Trial fish were randomly divided into the MBP0 group, MBP6 group, MBP12 group, and MBP18 group fed the diets with MBP replacing FM at levels of 0, 6%, 12%, and 18%, respectively. The trial lasted for ten weeks. There were no significant differences in weight gain or feed utilization among the MBP0, MBP6, and MBP12 groups (except for the feeding rate in the MBP12 group). Compared with the MBP0 group, the D-lactate level and diamine oxidase activity in the serum were significantly elevated in the MBP12 and MBP18 groups. In terms of non-specific immunity parameters in serum, the alkaline phosphatase activity was significantly decreased in the MBP18 group, and the complement 3 level was significantly elevated in the MBP12 and MBP18 groups. The activities of lipase and protease in the intestine were significantly decreased in the MBP12 and MBP18 groups. Compared with the MBP0 group, the total antioxidant capacity and activities of superoxide dismutase, catalase, and glutathione peroxidase in the intestine were significantly decreased in the MBP18 group, while the malondialdehyde level was significantly increased. The villus height, muscular thickness, and microvillus density were significantly decreased in the MBP12 and MBP18 groups. There were no significant differences in the foresaid parameters between the MBP0 group and the MBP6 group. The intestinal microbiota of the MBP6 group was beneficially regulated to maintain similar growth and health status with the MBP0 group. The adverse effects on the intestinal microbiota were reflected in the MBP18 group. In conclusion, MBP could successfully replace 6% of FM in the diet without adversely affecting the growth performance, serum biochemical parameters, and intestinal health of juvenile American eels. Full article

A 10-week growth study was conducted to evaluate the effect of a natural gas fermentation bacterial meal (FeedKind^®, FK) as a fishmeal (FM) alternative in largemouth bass (Micropterus salmoides) (48.0 ± 0.03 g). Four isonitrogenous and isoenergetic diets were formulated including one commercial control (C, 42% FM) and three experimental diets with gradient FK of 3% (FK3, 29%FM), 6% (FK6, 26%FM) and 9% (FK9, 23%FM), respectively. FK-fed groups showed significantly higher SR than that of C group. The WGR and SGR of fish fed FK3 and FK6 were significantly higher than those of FK9, but not statistical different from the C group. FK-fed groups showed higher apparent digestibility coefficients of dry matter and nutrients. Further, FK-fed groups increased the ratio of SOD/MDA in the plasma and liver, and the upregulation of intestinal Keap1 and downregulation of HIF1α was found in FK3. Furthermore, FK-fed groups showed higher microbial richness and diversity. Pearson correlation analysis found that antioxidant relevant biomarkers were negatively correlated with the relative abundance of certain potential beneficial bacteria. In conclusion, supplemented up to 3–6% FK replacing FM in a low FM diet of largemouth bass could increase growth, survival rate, antioxidant capacity, and improve gut microbiota. Full article

Candidatus Methylospira mobilis is a recently described spiral-shaped, micro-aerobic methanotroph, which inhabits northern freshwater wetlands and sediments. Due to difficulties of cultivation, it could not be obtained in a pure culture for a long time. Here, we report on the successful isolation of strain Shm1, the first axenic culture of this unique methanotroph. The complete genome sequence obtained for strain Shm1 was 4.7 Mb in size and contained over 4800 potential protein-coding genes. The array of genes encoding C₁ metabolic capabilities in strain Shm1 was highly similar to that in the closely related non-motile, moderately thermophilic methanotroph Methylococcus capsulatus Bath. The genomes of both methanotrophs encoded both low- and high-affinity oxidases, which allow their survival in a wide range of oxygen concentrations. The repertoire of signal transduction systems encoded in the genome of strain Shm1, however, by far exceeded that in Methylococcus capsulatus Bath but was comparable to those in other motile gammaproteobacterial methanotrophs. The complete set of motility genes, the presence of both the molybdenum–iron and vanadium-iron nitrogenases, as well as a large number of insertion sequences were also among the features, which define environmental adaptation of Methylospira mobilis to water-saturated, micro-oxic, heterogeneous habitats depleted in available nitrogen. Full article

Methane, an important greenhouse gas, has a 20-fold higher heat capacity than carbon dioxide. Earlier, through advanced spectroscopy and structural studies, the mechanisms underlying the extremely stable C–H activation of soluble methane monooxygenase (sMMO) have been elucidated in Methylosinus trichosporium OB3b and Methylococcus capsulatus Bath. Here, sMMO components—including hydroxylase (MMOH), regulatory (MMOB), and reductase (MMOR)—were expressed and purified from a type II methanotroph, Methylosinus sporium strain 5 (M. sporium 5), to characterize its hydroxylation mechanism. Two molar equivalents of MMOB are necessary to achieve catalytic activities and oxidized a broad range of substrates including alkanes, alkenes, halogens, and aromatics. Optimal activities were observed at pH 7.5 for most substrates possibly because of the electron transfer environment in MMOR. Substitution of MMOB or MMOR from another type II methanotroph, Methylocystis species M, retained specific enzyme activities, demonstrating the successful cross-reactivity of M. sporium 5. These results will provide fundamental information for further enzymatic studies to elucidate sMMO mechanisms. Full article