Transcriptomic profiling of Methylococcus capsulatus (Bath) during growth with two different methane monooxygenases.

Methylococcus capsulatus (Bath) is a methanotroph that possesses both a membrane-embedded (pMMO) and a soluble methane monooxygenase (sMMO). The expression of these two MMO's is tightly controlled by the availability of copper in the growth medium, but the underlying mechanisms and the number of genes involved in this switch in methane oxidation is not yet fully elucidated. Microarray analyses were used to assess the transcriptome in cells producing either pMMO or sMMO. A total of 137 genes were differentially expressed, with 87 genes showing a significant up-regulation during sMMO production. The majority of the differentially expressed genes could be assigned to functional roles in the energy metabolism and transport. Furthermore, three copper responding gene clusters were discovered, including an extended cluster that also harbors the genes for sMMO. Our data also indicates that major changes takes place in the respiratory chain between pMMO- and sMMO-producing cells, and that quinone are predominantly used as the electron donors for methane oxidation by pMMO. Intriguingly, a large proportion of the differentially expressed genes between pMMO- and sMMO-producing cells encode c-type cytochromes. By combining microarray- and mass spectrometry data, a total of 35 c-type cytochromes are apparently expressed in M. capsulatus when grown in nitrate mineral salt medium with methane as sole energy and carbon source, and the expression of 21 of these respond to the availability of copper. Interestingly, several of these c-type cytochromes are recovered from the cell surface, suggesting that extracellular electron transfers may occur in M. capsulatus.

The noncommensal bacterium Methylococcus capsulatus (Bath) ameliorates dextran sulfate (Sodium Salt)-Induced Ulcerative Colitis by influencing mechanisms essential for maintenance of the colonic barrier function.

Dietary inclusion of a bacterial meal has recently been shown to efficiently abolish soybean meal-induced enteritis in Atlantic salmon. The objective of this study was to investigate whether inclusion of this bacterial meal in the diet could abrogate disease development in a murine model of epithelial injury and colitis and thus possibly have therapeutic potential in human inflammatory bowel disease. C57BL/6N mice were fed ad libitum a control diet or an experimental diet containing 254 g/kg of body weight BioProtein, a bacterial meal consisting of Methylococcus capsulatus (Bath), together with the heterogenic bacteria Ralstonia sp., Brevibacillus agri, and Aneurinibacillus sp. At day 8, colitis was induced by 3.5% dextran sulfate sodium (DSS) ad libitum in the drinking water for 6 days. Symptoms of DSS treatment were less profound after prophylactic treatment with the diet containing the BioProtein. Colitis-associated parameters such as reduced body weight, colon shortening, and epithelial damage also showed significant improvement. Levels of acute-phase reactants, proteins whose plasma concentrations increase in response to inflammation, and neutrophil infiltration were reduced. On the other, increased epithelial cell proliferation and enhanced mucin 2 (Muc2) transcription indicated improved integrity of the colonic epithelial layer. BioProtein mainly consists of Methylococcus capsulatus (Bath) (88%). The results that we obtained when using a bacterial meal consisting of M. capsulatus (Bath) were similar to those obtained when using BioProtein in the DSS model. Our results show that a bacterial meal of the noncommensal bacterium M. capsulatus (Bath) has the potential to attenuate DSS-induced colitis in mice by enhancing colonic barrier function, as judged by increased epithelial proliferation and increased Muc2 transcription.

Bacteria grown on natural gas prevent soybean meal-induced enteritis in Atlantic salmon.

Dietary inclusion of solvent extracted soybean meal (SBM) is associated with inflammation in the distal intestine of salmonid fish, commonly referred to as SBM-induced enteritis. The enteritis is linked to alcohol soluble components in SBM, but the mechanisms have not been established. Previous studies show that bacterial meal (BM) containing mainly Methylococcus capsulatus grown on natural gas is a suitable protein source for salmonids. The BM is rich in nucleotides, phospholipids, and small peptides that might be beneficial for intestinal homeostasis. In this study, a fish meal (FM)-based control diet (FM diet) and diets with 200 g/kg SBM (SBM diet), 300 g/kg BM (BM diet), and 300 g/kg BM and 200 g/kg SBM (BM-SBM diet) were fed to juvenile Atlantic salmon (Salmo salar) for 80 d. Dietary inclusion of SBM reduced growth (P = 0.007). Inclusion of BM reduced digestibility of protein (P = 0.002) and lipids (P = 0.011) and increased (P < 0.01) the relative weights (g/kg whole body) of total gut, liver, and stomach, and mid and distal intestine. Fish fed the SBM diet developed enteritis, lacked carbonic anhydrase 12 in the brush border of epithelial cells in distal intestine, and had more epithelial cells reacting for proliferating cell nuclear antigen compared with fish fed the other diets. Fish fed the same amount of SBM combined with BM showed no signs of inflammation in the distal intestine. Our results demonstrate that BM grown on natural gas can be used to prevent SBM-induced enteritis in Atlantic salmon.

The RING-finger protein haprin: domains and function in the acrosome reaction.

The RBCC (RING finger, B-box type zinc finger, coiled-coil domain) motif family contains a large number of proteins implicated in many cellular processes, including vesicle exocytosis. The acrosome reaction, the sperm exocytotic event that is required for fertilization, involves essentially the same process of intracellular membrane fusions as vesicular exocytosis in somatic cells. We have previously isolated a haploid-germ-cell-specific gene designated haprin, which encodes a RBCC motif protein that plays a role in the acrosome reaction of sperm by mediating protein complex formation via the RBCC motif. In this review, we describe the potential role of Haprin in the molecular mechanisms of acrosome reaction, as compared with some other RBCC proteins. The conserved structure and localization of the Haprin protein in human and mouse suggest an indispensable role for Haprin in the functioning of mammalian sperm.