Alanine-scanning mutation approach for classification of the roles of conserved residues in the activity and substrate affinity of L-carnitine dehydrogenase.

L-Carnitine dehydrogenase (CDH) is as an excellent tool for L-carnitine (L-Car) estimation. To date, four CDHs have been identified, that share 45 % homology of their proteins. Here 42 conserved residues of CDH from Xanthomonas translucens (Xt-CDH) were substituted successively with alanine. The resultant mutants were analyzed for catalytic activity. Active mutants were evaluated for their influence on L-Car affinity. Twenty-three mutants with reduced affinity toward L-Car were subjected to detailed kinetic analysis. Analytical data implied that all mutants had increased K m values. The mutants of R193A, E196A, W199A, R200A, F249A, and F253A that produced the greatest L-Car affinity disruption (K m > 200-folds of Xt-CDH) clustered near the putative active site. This information can provide a solid basis for the rational design of mutagenic investigation to improve CDHs.

Root endophytic bacterial and fungal communities in a natural hot desert are differentially regulated in dry and wet seasons by stochastic processes and functional traits.

Dryland ecosystems experience seasonal cycles of severe drought and moderate precipitation. Desert plants may develop symbiotic relationships with root endophytic microbes to survive under the repeated wet and extremely dry conditions. Although community coalescence has been found in many systems, the colonization by functional microbes and its relationship to seasonal transitions in arid regions are not well understood. Here we examined root endophytic microbial taxa, and their traits in relation to their root colonization, during the dry and wet seasons in a hot desert of the southwestern United States. We used high-throughput DNA sequencing of 16S rRNA and internal transcribed spacer gene profiling of five desert shrubs, and analyzed the seasonal change in endophytic microbial lineages. Goodness of fit to the neutral community model in relationship to microbial traits was evaluated. In summer, Actinobacteria and Bacteroidia increased, although this was not genus-specific. For fungi, Glomeraceae selectively increased in summer. In winter, Gram-negative bacterial genera, including those capable of nitrogen fixation and plant growth promotion, increased. Neutral model analysis revealed a strong stochastic influence on endophytic bacteria but a weak effect for fungi, especially in summer. The taxa with higher frequency than that predicted by neutral model shared environmental adaptability and symbiotic traits, whereas the frequency of pathogenic fungi was at or under the predicted value. These results suggest that community assembly of bacteria and fungi is regulated differently. The bacterial community was affected by stochastic and deterministic processes via bacterial response to drought (response trait), beneficial effect on plants (effect trait), and likely stable mutualistic interactions with plants suggested by the frequency of nodule bacteria. For fungi, mycorrhizal fungi were selected by plants in summer. The regulation of beneficial microbes by plants in both dry and wet seasons suggests the presence of plant-soil positive feedback in this natural desert ecosystem.

Chemicals behind the use of Strombus tricornis opercula in traditional sudanese perfumery and medicine.

Strombidae is one of the major molluscan families in Sudan and due to their opercula, has tremendous economic value. In traditional Sudanese homemade perfumes and body care cosmetics, Strombidae family operculum is one of the main ingredients. Their fumigation generates a charming odor preferred by Sudanese people, used for body smoke baths by married women. Moreover, these fumes are believed to treat several gynecological disorders. In this study, we attempted to confirm the presence of volatiles with pleasant odors and compounds with pharmaceutical importance in the Strombidae opercula. Volatiles from the smoke and soak extracts of the burned opercula were analyzed using gas chromatography-mass spectrometry (GC-MS). Furthermore, polar components from the methanol extract of opercula powder were isolated using high-performance liquid chromatography (HPLC) and identified by nuclear magnetic resonance (NMR), electrospray ionization mass spectrometry (ESI-MS), and UV spectra. The elemental and metal contents were analyzed using inductively coupled plasma-mass spectrometry (ICP-MS). GC-MS analysis revealed several phenols, aldehydes, ketones, and other functional fragrant and volatile constituents. Further, two compounds were purified from the methanol extract of Strombidae opercula, and named compounds B and D, which were identified as cyclo-(Tyr-Gly) and 4-hydroxybenzaldehyde, respectively. ICP-MS analysis revealed the presence of various elements and metals at different levels. These findings support the historical and traditional practices and usage of the Strombidae opercula in therapeutic and esthetic products. The opercula contains many biologically active compounds and produces smoke containing volatile scent compounds, which might provide alternative pharmaceuticals and cosmetic ingredients that can cooperate to improve the manufacturing of numerous medical products.

Argeloside I inhibits the pathogenicity of Porphyromonas gingivalis TDC60.

Porphyromonas gingivalis, a major pathogen associated with chronic periodontitis, produces several virulence agents in the outer cell membrane, including gingipains and hemagglutinins. These virulence factors enable the bacteria to adhere to periodontal tissue and degrade host proteins to obtain the nutrients needed for dental plaque formation. P. gingivalis TDC60 was recently identified as the most aggressive P. gingivalis strain to dates. In this study, we isolated a known pregnane glycoside, argeloside I, from the aqueous extract of Solenostemma argel leaves. Argeloside I completely hindered the growth of P. gingivalis TDC60 and inhibited the production of hemagglutinins as well as Arg- and Lys-specific gingipains. Our results demonstrate a new function of pregnane glycosides. Argeloside I may be a candidate for reducing the risk associated with P. gingivalis TDC60 and its adhesion factors.

Isolates from Monechma ciliatum seeds' extract hampered Porphyromonas gingivalis hemagglutinins.

Porphyromonas gingivalis is a major periodontitis pathogen that produces several virulence factors including hemagglutinins. These proteins, which are vital molecules, allow P. gingivalis to uptake iron and heme by attaching, aggregating, and lysing erythrocytes. In this study, we evaluated the inhibitory activity of the aqueous extract of Monechma ciliatum seeds against the hemagglutination activity of P. gingivalis. M. ciliatum is a Sudanese medicinal herb that grows in arid and semi-arid lands of tropical Africa. The water extracted from dry powdered seeds was partitioned using ethyl acetate followed by reversed-phase chromatography, thin-layer chromatography, ESI-MS, and NMR analysis resulting in the isolation of four compounds identified as oleic acid, coumarin, 1,2-dioleoylglycerol, and 1,3-dioleoylglycerol with MICs of 15-100 µg/ml against hemagglutination. We believe that the isolation and purification of these compounds will expand the application of M. ciliatum as a natural therapeutic or preventative agent. PRACTICAL APPLICATIONS: Monechma ciliatum or black mahlab is a famous medicinal plant that grows in some parts of arid and semi-arid areas of tropical Africa including western Sudan. Despite its nutritional and traditional medical applications, no studies have evaluated its anti-hemagglutination activity against periodontal pathogens. In this study, four active compounds (oleic acid, coumarin, 1,2-dioleoylglycerol, and 1,3-dioleoylglycerol) were isolated and identified from an aqueous extract of M. ciliatum seeds. The isolated compounds revealed high levels of inhibitory activity against all hemagglutinin agents secreted by Porphyromonas gingivalis. This evidence of inhibitory activity will encourage the application of M. ciliatum effectively as a functional food or therapeutic agent to prevent periodontal diseases in the early stages.

Chemical composition, antioxidant capacity, and mineral extractability of Sudanese date palm (Phoenix dactylifera L.) fruits.

The aim of the present work was to investigate the chemical composition, mineral extractability, and antioxidant capacity of six date palm varieties grown in Sudan. The results showed that Sudanese date varieties contained significantly different (P < 0.05) amounts of moisture, ash, fiber, oil, and carbohydrates, but have almost similar amounts of protein. Moreover, results revealed that date varieties contained significantly varied (P < 0.05) amounts of total polyphenols and total flavonoids, which ranged between 35.82 and 99.34 mg gallic acid equivalent/100 g and 1.74-3.39 mg catechin equivalent/100 g, respectively. The antioxidant activities of the studied date varieties were as follows: ferric-reducing antioxidant power (FRAP) was within the range of 2.82-27.5 mmol/100 g, chelation of Fe(2+) ion ranged from 54.31% to 94.98%, and scavenging of H2O2 ranged from 38.48% to 49.13%. There were many correlations (positive, negative, and weak) between antioxidant and mineral extractability of Sudanese date fruits.

Identification of residues essential for the activity and substrate affinity of L-carnitine dehydrogenase.

Recently, two L-carnitine dehydrogenases from soil isolates Rhizobium sp. (Rs-CDH) and Xanthomonas translucens (Xt-CDH) have demonstrated to exhibit mutually differing affinities toward L-carnitine. To identify residues important for affinity to the substrate, we compared the primary structure of Xt-CDH and Rs-CDH with the recognized 3D structure of 3-hydroxyacyl-CoA dehydrogenase (PDB code: 1F0Y). Then, six residues of Xt-CDH (Phe143, Gly188, Ile190, Ala191, Gly223, and Ala224) and the corresponding residues of Rs-CDH (Tyr140, Ala185, Val187, Gly188, Ser220, and Phe221) were selected for further mutagenesis. The residues of Xt-CDH were replaced with that of Rs-CDH at the corresponding position and vice versa. All Rs-CDH mutants exhibited slight effects on substrate affinity, except for the double mutants Rs-V187I/G188A, which was devoid of enzyme activity. All Xt-CDH mutants showed different K m values. Xt-F143Y caused a higher increase in the K m value. Furthermore, the kinetic parameters of 10 mutants at Xt-F143 and Rs-Y140 were investigated. All Rs-Y140 mutants, except aromatic residues (Phe, Trp), produced proteins that were almost entirely devoid of enzyme activity and with disrupted affinity to L-carnitine. All Xt-F143 variants showed a marked reduction (P ≤ 0.05) in enzyme activity. Overall, our results suggest that the aromatic rings of Tyr140 in Rs-CDH and Phe143 of Xt-CDH are essential for substrate recognition.

Metal and metalloid bioaccumulation in the Pacific blue shrimp Litopenaeus stylirostris (Stimpson) from New Caledonia: laboratory and field studies.

The present work aimed at better understanding metal and metalloid bioaccumulation in the edible Pacific blue shrimp Litopenaeus stylirostris, using both laboratory and field approaches. In the laboratory, the bioaccumulation kinetics of Ag, Cd, Co, Cr, and Zn have been investigated in shrimp exposed via seawater and food, using the corresponding gamma-emitting radiotracers ((110 m)Ag, (109)Cd, (57)Co, (51)Cr, and (65)Zn) and highly sensitive nuclear detection techniques. Results showed that hepatopancreas and intestine concentrated the metals to the highest extent among the blue shrimp organs and tissues. Moulting was found to play a non negligible detoxification role for Co, Cr and, to a lesser extent, Zn. Metal retention by L. stylirostris widely varied (from a few days to several months), according to the element and exposure pathway considered (a given metal was usually less strongly retained when ingested with food than when it was taken up from the dissolved phase). In the field study, Ag, As, Cd, Co, Cr, Cu, Mn, Ni, and Zn were analysed in shrimp collected from a New Caledonian aquaculture pond. Metal concentrations in the shrimp muscles were generally relatively low and results confirmed the role played by the digestive organs and tissues in the bioaccumulation/storage/detoxification of metals in the Pacific blue shrimp. Preliminary risk considerations indicate that consumption of the shrimp farmed in New Caledonia is not of particular concern for human health.