33 research outputs found
Influence of Substrates on the Surface Characteristics and Membrane Proteome of Fibrobacter succinogenes S85
Although Fibrobacter succinogenes S85 is one of the most proficient cellulose degrading bacteria among all mesophilic organisms in the rumen of herbivores, the molecular mechanism behind cellulose degradation by this bacterium is not fully elucidated. Previous studies have indicated that cell surface proteins might play a role in adhesion to and subsequent degradation of cellulose in this bacterium. It has also been suggested that cellulose degradation machinery on the surface may be selectively expressed in response to the presence of cellulose. Based on the genome sequence, several models of cellulose degradation have been suggested. The aim of this study is to evaluate the role of the cell envelope proteins in adhesion to cellulose and to gain a better understanding of the subsequent cellulose degradation mechanism in this bacterium. Comparative analysis of the surface (exposed outer membrane) chemistry of the cells grown in glucose, acid-swollen cellulose and microcrystalline cellulose using physico-chemical characterisation techniques such as electrophoretic mobility analysis, microbial adhesion to hydrocarbons assay and Fourier transform infra-red spectroscopy, suggest that adhesion to cellulose is a consequence of an increase in protein display and a concomitant reduction in the cell surface polysaccharides in the presence of cellulose. In order to gain further understanding of the molecular mechanism of cellulose degradation in this bacterium, the cell envelope-associated proteins were enriched using affinity purification and identified by tandem mass spectrometry. In total, 185 cell envelope-associated proteins were confidently identified. Of these, 25 proteins are predicted to be involved in cellulose adhesion and degradation, and 43 proteins are involved in solute transport and energy generation. Our results supports the model that cellulose degradation in F. succinogenes occurs at the outer membrane with active transport of cellodextrins across for further metabolism of cellodextrins to glucose in the periplasmic space and inner cytoplasmic membrane
The Complete Genome Sequence of Fibrobacter succinogenes S85 Reveals a Cellulolytic and Metabolic Specialist
Fibrobacter succinogenes is an important member of the rumen
microbial community that converts plant biomass into nutrients usable by its
host. This bacterium, which is also one of only two cultivated species in its
phylum, is an efficient and prolific degrader of cellulose. Specifically, it has
a particularly high activity against crystalline cellulose that requires close
physical contact with this substrate. However, unlike other known cellulolytic
microbes, it does not degrade cellulose using a cellulosome or by producing high
extracellular titers of cellulase enzymes. To better understand the biology of
F. succinogenes, we sequenced the genome of the type strain
S85 to completion. A total of 3,085 open reading frames were predicted from its
3.84 Mbp genome. Analysis of sequences predicted to encode for
carbohydrate-degrading enzymes revealed an unusually high number of genes that
were classified into 49 different families of glycoside hydrolases, carbohydrate
binding modules (CBMs), carbohydrate esterases, and polysaccharide lyases. Of
the 31 identified cellulases, none contain CBMs in families 1, 2, and 3,
typically associated with crystalline cellulose degradation. Polysaccharide
hydrolysis and utilization assays showed that F. succinogenes
was able to hydrolyze a number of polysaccharides, but could only utilize the
hydrolytic products of cellulose. This suggests that F.
succinogenes uses its array of hemicellulose-degrading enzymes to
remove hemicelluloses to gain access to cellulose. This is reflected in its
genome, as F. succinogenes lacks many of the genes necessary to
transport and metabolize the hydrolytic products of non-cellulose
polysaccharides. The F. succinogenes genome reveals a bacterium
that specializes in cellulose as its sole energy source, and provides insight
into a novel strategy for cellulose degradation
The effect of groove texture patterns on piston-ring pack friction
A cylinder liner possesses fairly intricate surface requirements due to its complicated functions. It needs to provide adequate surface roughness to resist wear as well as to store and retain lubricants during high temperatures. The liner surface texture is anisotropic, produced by the honing process, with resultant deep visible scratches left on it [1]. The prominence of the honing grooves observed suggests that surface texture significantly affects ring-pack performance, although this effect is not clearly understood. In this paper, a numerical model was developed to investigate the effects of groove characteristics on the lubrication condition and friction at the interface between the piston ring and cylinder liner. This model aims to solve the average Reynolds equation, which depends on the real surface topographies of the cylinder liner, and describes the influence of surface irregularities on the lubricant flow under hydrodynamic lubrication conditions, considering lubricant film rupture and cavitations. Numerical results help to determine the optimum lateral groove characteristics to reduce friction and then noxious emissions