Molecular genetics of Streptococcus thermophilus

Abstract The metabolism and genetics of Streptococcus thermophilus (presently Streptococcus salivarius ssp. thermophilus ) have only been investigated recently despite its widespread use in milk fermentation processes. The development of recombinant DNA technology has allowed impressive progress to be made in the knowledge of thermophilic dairy streptococci. In particular, it has permitted a careful analysis of phenotypically altered variants which were derived from a mother strain by plasmid or chromosomal DNA reorganization. While natural phage defense mechanisms of S. thermophilus remain poorly documented, information on the bacteriophages responsible for fermentation failures has accumulated. The lysogenic state of two S. thermophilus strains has also been demonstrated for the first time. Gene transfer techniques for this species have been established and improved to the point that targeted manipulation of their chromosomal determinants is now feasible. Cloning and expression vectors have been constructed, and a few heterologous genes were successfully expressed in S. thermophilus . The first homologous genes, involved in carbohydrate utilization, have been cloned and sequenced, shedding some light on the molecular organization of key metabolic steps.     

Lactate has the potential to promote hydrogen sulphide formation in the human colon

High concentrations of sulphide are toxic for the gut epithelium and may contribute to bowel disease. Lactate is a favoured cosubstrate for the sulphate-reducing colonic bacterium Desulfovibrio piger , as shown here by the stimulation of sulphide formation by D. piger DSM749 by lactate in the presence of sulphate. Sulphide formation by D. piger was also stimulated in cocultures with the lactate-producing bacterium Bifidobacterium adolescentis L2-32. Other lactate-utilizing bacteria such as the butyrate-producing species Eubacterium hallii and Anaerostipes caccae are, however, expected to be in competition with the sulphate-reducing bacteria (SRB) for the lactate formed in the human colon. Strains of E. hallii and A. caccae produced 65% and 96% less butyrate from lactate, respectively, in a coculture with D. piger DSM749 than in a pure culture. In triculture experiments involving B. adolescentis L2-32, up to 50% inhibition of butyrate formation by E. hallii and A. caccae was observed in the presence of D. piger DSM749. On the other hand, sulphide formation by D. piger was unaffected by E. hallii or A. caccae in these cocultures and tricultures. These experiments strongly suggest that lactate can stimulate sulphide formation by SRB present in the colon, with possible consequences for conditions such as colitis.     

Pectic Metabolism in Suspension-Cultured Cells of Flax, Linum usitatissimum

Flax cells in suspension culture continuously synthesize twotypes of pectin (i) highly methylated pectin that is covalentlylinked to the cell wall (ii) and low methylated pectin thatis ionically linked to the cell wall. A maximal activity ofpectin methyltransferase (PMT) in microsomes was recorded atthe beginning of the elongation phase, while, two maximal activitiesof pectin methylesterase (PME) in cell walls were recorded atthe end of the elongation phase and during the maturation phase.The differences between the PMT and PME activities, not onlyin terms of their location but also in terms of the time courseof the culture, may explain the variations in the cation-exchangecapacity (CEC) of the cell walls, which is mainly due to thenegative charges of the less methylated pectins. The minimalvalue of the CEC during the elongation phase was due to high-levelsynthesis of neutral, hemicellulose-like polysaccharides andalso to the PMT activity into the highly methylated, covalentlylinked pectins. Conversely, the large increase of the CEC duringthe maturation phase, might be due to the low level of PMT activitythat is responsible for the limited methylation of the pectinssynthesized at that time. (Received November 2, 1992; Accepted June 14, 1993)     

Yield performance of 14 novel inter- and intra-species Miscanthus hybrids across Europe

Miscanthus, a C4 perennial rhizomatous grass from Asia is a leading candidate for the supply of sustainable biomass needed to grow the bioeconomy. European Miscanthus breeding programmes have recently produced a new range of seeded hybrids with the objective of increasing scalability to large acreages limited by current clonal propagation. For the EU-GRACE project, new replicated field trials were established in seven locations across Europe in 2018 with eight intraspecific M. sinensis hybrids (sin × sin) and six M. sacchariflorus × M. sinensis (sac × sin) from Dutch and UK breeding programmes, respectively, with clonal Miscanthus × giganteus. The planting density of the sin × sin was double that of sac × sin (30,000 & 15,000 plants ha⁻¹), creating commercially relevant upscaling comparisons between systems. Over the first 3 years, the establishment depended on location and hybrid. The mature sin × sin hybrids formed tight tufts of shoots up to 2.5 m tall which flower and senesce earlier than the taller sac × sin hybrids. Following the third growing season, the highest yields were recorded in Northern Italy at a low altitude (average 13.7 (max 21) Mg DM ha⁻¹) and the lowest yielding was on the industrially damaged marginal land site in Northern France (average 7.0 (max 10) Mg DM ha⁻¹). Moisture contents at spring harvest were lowest in Croatia (21.7%) and highest in Wales, UK (41.6%). Overall, lower moisture contents at harvest, which are highly desirable for transport, storage and for most end-use applications, were found in sin × sin hybrids than sac × sin (30% and 40%, respectively). Yield depended on climate interactions with the hybrid and their associated planting systems. The sin × sin hybrids appeared better adapted to northern Europe and sac × sin hybrids to southern Europe. Longer-term yield observations over crop lifespans will be needed to explore the biological (yield persistence) and economic costs and benefits of the different hybrid systems.     

Lectin Receptor Kinases in Plants

Referee: Dr. Philip Becraft, Zoology and Genetics/Agronomy Depts., 2116 Molecular Building, lowa State University, Ames, IA 50011 Forty-two lectin receptor kinase (lecRK)-related sequences and nine related soluble legume lectin sequences were identified in the Arabidopsis thaliana genome. The genes are scattered as a single or gathered copies at different loci throughout the five chromosomes, and four predicted lecRK probably correspond to pseudogenes. Both structural alignments and molecular modeling revealed striking similarities between the lectinlike domain of lecRK, and related A. thaliana soluble lectins and legume lectins. The hydrophobic cavity is extremely conserved, whereas most of the residues forming the monosaccharide-binding site and the bivalent cation-binding site of legume lectins are poorly conserved. LecRK should be unable to bind the simple sugars usually recognized by genuine legume lectins. Molecular modeling of the kinase domain suggests that, except for two apparently inactive receptors, all other lecRK contain a putative functional Ser/Thr kinase catalytic domain. Both the juxtamembrane and C-terminal domains, which are considered important regions for regulating the kinase activity, exhibit a few specific stretches of amino acid residues. Some phylogenetic relationships are inferred from the phylogenetic trees built up from the different lecRK domain sequences. LecRK cluster in three distinct classes (A,B,C), one of them (B) being more closely related to soluble lectins of A. thaliana and legume lectins.     

Multiple Human Taste Receptor Sites: A Molecular Modeling Approach

Numerous experimental data on the human peripheral taste systemsuggest the existence of multiple low-affinity and low-specificityreceptor sites which are responsible for the detection and thecomplete discrimination of a very large number of organic molecules.According to this hypothesis, a given molecule interacts withnumerous taste receptors and vice versa. Statistical analysisof taste intensities estimated by 58 human subjects for variousmolecules enables the calculation of taste intermolecular distances.For the present modeling study, we hypothesized that a shorttaste distance (i.e. taste similarity) between two distinctmolecules indicates that they bind with similar distributionsof affinities to the taste receptors, and hence display similarbinding motifs. In order to find common molecular binding motifsamong 14 selected organic tastants, hydrogen-bonding and hydrophobicinteraction properties were mapped onto their molecular surfaces.The 14 surfaces were then cut in 240 fragments, most of whichwere made up of 2–4 potentially interacting zones. A correspondenceindex was defined to measure the analogy between two optimallysuperimposed fragments. The 75 most representative fragmentswere all matched pairwise. Twelve distinct clusters of fragmentswere isolated from the 2775 calculated comparisons. These 12fragment types were used to calculate structural similaritydistances. We then performed a combinatorial analysis to identifywhich fragment combination best reconciled structural and tastedistances. We finally identified an optimal subset of sevenfragment types out of the 12, which significantly and best accountedfor the 91 pairwise taste distances between all 14 modeled tastants.These seven validated fragment types are therefore presentedas good candidates to be recognized by the same number of distincttaste receptor sites. Potential applications of these identifiedbinding motifs to tastant design are suggested. Chem. Senses21: 425–445, 1996.     

Mortality Rates above Emergency Threshold in Population Affected by Conflict in North Kivu,Democratic Republic of Congo,July 2012–April 2013

The area of Walikale in North Kivu, Democratic Republic of Congo, is intensely affected by conflict and population displacement. Médecins-Sans-Frontières (MSF) returned to provide primary healthcare in July 2012. To better understand the impact of the ongoing conflict and displacement on the population, a retrospective mortality survey was conducted in April 2013. A two-stage randomized cluster survey using 31 clusters of 21 households was conducted. Heads of households provided information on their household make-up, ownership of non-food items (NFIs), access to healthcare and information on deaths and occurrence of self-reported disease in the household during the recall period. The recall period was of 325 days (July 2012–April 2013). In total, 173 deaths were reported during the recall period. The crude mortality rate (CMR) was of 1.4/10,000 persons/day (CI95%: 1.2–1.7) and the under-five- mortality rate (U5MR) of 1.9/10,000 persons per day (CI95%: 1.3–2.5). The most frequently reported cause of death was fever/malaria 34.1% (CI95%: 25.4–42.9). Thirteen deaths were due to intentional violence. Over 70% of all households had been displaced at some time during the recall period. Out of households with someone sick in the last two weeks, 63.8% sought health care; the main reason not to seek health care was the lack of money (n = 134, 63.8%, CI95%: 52.2–75.4). Non Food Items (NFI) ownership was low: 69.0% (CI95%: 53.1–79.7) at least one 10 liter jerry can, 30.1% (CI95%: 24.3–36.5) of households with visible soap available and 1.6 bednets per household. The results from this survey in Walikale clearly illustrate the impact that ongoing conflict and displacement are having on the population in this part of DRC. The gravity of their health status was highlighted by a CMR that was well above the emergency threshold of 1 person/10,000/day and an U5MR that approaches the 2 children/10,000/day threshold for the recall period.