Isolation of a seed coagulant Moringa oleifera lectin

In this work hemagglutinating activity (HA) was investigated in distinct Moringa oleifera tissue extracts. A new lectin from seeds (cMoL) was purified and characterized; hemagglutinating and coagulating activities were evaluated. HA was detected in 0.15 M NaCl extracts from flowers and rachis inflorescence (5%, w/v), seeds, leaves, fundamental tissue of stem and steam bark (10%, w/v). cMoL isolated after saline extraction and guar gel column chromatography was active at pH range 4.0–9.0 agglutinating erythrocytes from rabbit and human blood types. Extracts of tissues and cMoL activities were carbohydrate inhibited; azocasein and asialofetuin abolished cMoL HA. The lectin was thermostable at 100 °C during 7 h. Polyacrylamide gel electrophoresis under reduced conditions revealed a main polypeptide band of 26.5 kDa; native basic cMoL was detected as a unique band. Seed lectin preparations and cMoL showed coagulant activity, similar to aluminium sulphate, the coagulant most widely used in water treatment.     

Single-step ion exchange purification of the coagulant protein from Moringa oleifera seed

The coagulant protein from Moringa oleifera (MO) seed was purified using a single-step batch ion exchange (IEX) method. Adsorption and elution parameters were optimized. Impact of the purification on the reduction of organic and nutrient release to the water was studied. The matrix was equilibrated using ammonium acetate buffer, and the optimum ionic strength of NaCl for elution was 0.6 M. The time for adsorption equilibrium was between 90 and 120 min. Maximum adsorption capacity of the matrix, estimated with the Langmuir model, was 68 mg protein/g adsorbent. The purified protein does not release organic and nutrient loads to the water, which are the main concerns of the crude extract. This work suggests that a readily scalable single-step IEX purification method can be used to produce the coagulant protein and it can be carried out with locally available facilities. This will promote the use of MO in large water treatment plants and other industries.     

Floral Development and Anatomy ofMoringa oleifera(Moringaceae): What is the Evidence for a Capparalean or Sapindalean Affinity?

Floral development and anatomy ofMoringahave been investigatedin the context of the disputed view of a capparalean affinity.Flowers arise in terminal or axillary panicles. Sepals arisesequentially and petals simultaneously. Antepetalous stamensarise simultaneously and precede the antesepalous staminodes,which emerge sequentially. Within their respective whorls, thepetals and stamens become twisted along different orientations.The gynoecium develops as a ring primordium on which three carpellarylobes become demarcated simultaneously. A saccate ovary bearsnumerous ovules on a parietal placentation and is topped bya hollow style. The interpretation of laminal placentation isdenied. Monothecal anthers are formed by the failure of onehalf to initiate. The flowers present a peculiar form of zygomorphyrunning transversally from the petal between sepals 3 and 5to sepal 4. The shape and position of petals and stamens isrelated to a pollen presentation mechanism with bowl-shapedanthers on different levels. The floral anatomy also reflectsthe zygomorphy of the flower. AlthoughMoringashares importantmorphological features with certain members of the Sapindalesand Capparales, differences in ontogeny make a close relationshipwith either Capparales or certain Sapindales appear uncertain.Copyright1998 Annals of Botany Company Moringa,Moringaceae, Capparales, Sapindales, floral ontogeny, floral anatomy.     

Effects of storage conditions of Moringa oleifera seeds on its performance in coagulation

Moringa oleifera is a plant whose seeds have coagulation properties for treating water and wastewater. In this study the coagulation efficiency of Moringa oleifera kept in different storage conditions were studied. The Moringa oleifera seeds were stored at different conditions and durations; open container and closed container at room temperature (28 degrees C) and refrigerator (3 degrees C) for durations of 1, 3 and 5 months. Comparison between turbidity removal efficiency of Moringa oleifera kept in refrigerator and room temperature revealed that there was no significant difference between them. The Moringa oleifera kept in refrigerator and room temperature for one month showed higher turbidity removal efficiency, compared to those kept for 3 and 5 months, at both containers. The coagulation efficiency of Moringa oleifera was found to be dependent on initial turbidity of water samples. Highest turbidity removals were obtained for water with very high initial turbidity. In summary coagulation efficiency of Moringa oleifera was found independent of storage temperature and container, however coagulation efficiency of Moringa oleifera decreased as storage duration increased. In addition, Moringa oleifera can be used as a potential coagulant especially for very high turbidity water.     

Cytological and morphology characteristics of natural microsporogenesis within Camellia oleifera

Camellia oleifera is believed to exhibit a complex intraspecific polyploidy phenomenon. Abnormal microsporogenesis can promote the formation of unreduced gametes in plants and lead to sexual polyploidy, so it is hypothesized that improper meiosis probably results in the formation of natural polyploidy in Camellia oleifera. In this study, based on the cytological observation of meiosis in pollen mother cells (PMCs), we found natural 2n pollen for the first time in Camellia oleifera, which may lead to the formation of natural polyploids by sexual polyploidization. Additionally, abnormal cytological behaviour during meiosis, including univalent chromosomes, extraequatorial chromosomes, early segregation, laggard chromosomes, chromosome stickiness, asynchronous meiosis and deviant cytokinesis (monad, dyads, triads), was observed, which could be the cause of 2n pollen formation. Moreover, we confirmed a relationship among the length–width ratio of flower buds, stylet length and microsporogenesis. This result suggested that we can immediately determine the microsporogenesis stages by phenotypic characteristics, which may be applicable to breeding advanced germplasm in Camellia oleifera.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-01002-5.     

The efficiency of combined coagulant and ballast to remove harmful cyanobacterial blooms in a tropical shallow system

We tested the hypothesis that a combination of coagulant and ballast could be efficient for removal of positively buoyant harmful cyanobacteria in shallow tropical waterbodies, and will not promote the release of cyanotoxins. This laboratory study examined the efficacy of coagulants [polyaluminium chloride (PAC) and chitosan (made of shrimp shells)] alone, and combined with ballast (lanthanum modified bentonite, red soil or gravel) to remove the natural populations of cyanobacteria collected from a shallow eutrophic urban reservoir with alternating blooms of Cylindrospermopsis and Microcystis. PAC combined with ballast was effective in settling blooms dominated by Microcystis or Cylindrospermopsis. Contrary to our expectation, chitosan combined with ballast was only effective in settling Cylindrospermopsis-dominated blooms at low pH, whereas at pH ≥ 8 no effective flocculation and settling could be evoked. Chitosan also had a detrimental effect on Cylindrospermopsis causing the release of saxitoxins. In contrast, no detrimental effect on Microcystis was observed and all coagulant-ballast treatments were effective in not only settling the Microcystis dominated bloom, but also lowering dissolved microcystin concentrations. Our data show that the best procedure for biomass reduction also depends on the dominant species.     

Carbon isotope discrimination,water-use efficiency,growth, and mortality in a natural shrub population

In order to scale up from the ecophysiological characters of individual plants to population-level questions, we need to determine if character patterns in natural populations are stable through time, and if the characters are related to growth and survival. We investigated these questions in a 3-year study for one character, integrated water-use efficiency (WUE) as estimated by carbon isotope discrimination () in a population of the Great Basin shrub, Chrysothamnus nauseosus. WUE was a conservative character for a given plant within and across seasons, and a previously documented difference between two size classes (represented by juveniles and adults) was maintained; smaller juveniles had a lower WUE than larger adults. The lower WUE of juveniles was often accompanied by higher rates of photosynthesis and stomatal conductance as compared to adults even though juveniles generally had more negative xylem pressure potentials. Although many discussions of the role of WUE in natural populations have been based on the expectation that higher WUE (lower ) is generally associated with less growth, we found no such relation-ship for juvenile plants in this population (i.e was not positively correlated with height increase). In addition, juvenile plant mortality was not correlated with . Although there were stable patterns of WUE for plants in this population, the positive correlation between WUE and size, and the lack of a negative correlation between WUE and height growth, make it unlikely that the WUE of an individual plant will be related in a simple manner to its growth and survival in the population.     

Recombinant protein expression of Moringa oleifera lectin in methylotrophic yeast as active coagulant for sustainable high turbid water treatment

The natural coagulant Moringa oleifera lectin (MoL) as cationic protein is a promising candidate in coagulation process of water treatment plant. Introducing the gene encoding MoL into a host, Pichia pastoris, to secrete soluble recombinant protein is assessed in this study. Initial screening using PCR confirmed the insertion of MoL gene, and SDS-PAGE analysis detected the MoL protein at 8 kDa. Cultured optimization showed the highest MoL protein at 520 mg/L was observed at 28 °C for 144 h of culturing by induction in 1% methanol. Approximately, 0.40 mg/mL of recombinant MoL protein showed 95 ± 2% turbidity removal of 1% kaolin suspension. In 0.1% kaolin suspension, the concentration of MoL at 10 μg/mL exhibits the highest turbidity reduction at 68 ± 1%. Thus, recombinant MoL protein from P. pastoris is an effective coagulant for water treatment.     

江西省油茶综合生产潜力与资源利用效率评估

江西适合油茶生长,但开发利用程度不高,分析评估其生产潜力与资源利用效率,可为进一步优化油茶种植分区、提高产量、合理利用国土资源等提供依据。以江西省油茶综合生产潜力为研究对象,基于潜力衰减法逐级修订得到自然生产潜力,结合社会有效系数得到综合生产潜力,并进行资源利用效率评估。结果表明: 1)江西省油茶各级生产潜力平均值在10229～17724 kg·km^-2,从赣南向赣西北递减,平均综合生产潜力为12550 kg·km^-2,分级后具有较高潜力和高潜力类可利用土地面积达51656 km²,占全省面积的31.1%;市级生产潜力中,赣州最高。2)江西省油茶生产潜力变幅为2223～3857 kg·km^-2,各级潜力变化量的区域差异较小;油茶资源满足率为69.3%～122.7%,各级资源满足率区域差异较小;油茶资源组合利用效率为49.1%～85%,各级生产潜力利用率的区域差异较大。3)综合生产潜力评估结果与江西油茶实际产量较符合,资源利用效率评估结果与江西自然和社会条件相符,可以作为相关研究与政策制定的参考。     

Ethylene removal efficiency and bacterial community diversity of a natural zeolite biofilter

To establish an economical and environmentally friendly technology for ethylene removal from horticultural facilities and industrial point sources, a bench-scale natural zeolite biofiltration system was developed in this study. The system was evaluated for its performance in removing ethylene from an artificially contaminated air stream and characterized for its bacterial diversity under varied ethylene concentrations, and in different spatial stages of the filter. The biofilter enabled to approximately 100% remove ethylene at loading rates of 0.26-3.76 g m⁻³ h⁻¹ when operated with inoculum containing enriched ethylene-degrading bacteria. The bacterial diversity and abundance varied with the height of the biofilter. Moreover, the occurrence and predominance of specific bacterial species varied with the concentrations of ethylene introduced into the biofilter, as observed by PCR-DGGE methods. Phylogenetic analysis indicated that the biofilter system supported a diverse community of ethylene-degrading bacteria, with high similarity to species in the classes Betaproteobacteria, Gammaproteobacteria, Bacilli, and Actinobacteria.     

Preservation of organic-walled dinoflagellate cysts in different oxygen regimes: a 10,000 year natural experiment

The occurrence of organic-walled dinoflagellate cysts in (fossil) sediments depends on several factors, including as the ecological preferences of the cyst-forming dinoflagellates, cyst production, transport and preservation. Although laboratory experiments have shown that several cyst species are sensitive to chemical treatment, no information about the selective preservation of dinoflagellate cyst species in natural environments has previously been presented. Here, we present data on the effects of oxygen availability in bottom sediments on a cyst assemblage from the ungraded Madeira Abyssal Plain f-turbidite of which only the upper layer has been oxidized. Based on differences in species composition between the oxidized and underlying, unoxidized layers of this turbidite, the influence of oxygen availability on the preservation of individual species has been estimated. Cyst species have been classified in ascending order of resistance to oxygen availability in sediments as: (1) highly sensitive (cysts formed by Protoperidinium species), (2) moderately sensitive (e.g. Spiniferites species), (3) moderately resistant (e.g. Impagidinium paradoxum and Nematosphaeropsis labyrinthus) and (4) resistant (e.g. Impagidinium aculeatum).     

Density-dependent natural selection does not increase efficiency

Summary Populations ofDrosophila melanogaster kept at high population density (K-selected) for 125 generations have higher larval viability than populations kept at low densities (r-selected) when both are raised under crowded conditions. In additionK-selected adults that emerge from crowded cultures are larger than theirr-selected counterparts. These differences cannot be explained by differences in efficiency of food use. The minimum food required for successful pupation is actually greater in theK-selected populations. I conjecture that there may be a trade-off between minimum food requirements and competitive ability, which has changed substantially in theK-selected populations. The possibility thatK-selected larvae can dig more more deeply and gain access to unused food is examined and rejected as a possible explanation of the viability differences. Evidence is provided supporting the hypothesis that the differences in viability may be due to an increased tendency of theK-selected larvae to pupate off the surface of the medium.     

Calumenin knockdown,by intronic artificial microRNA,to improve expression efficiency of the recombinant human coagulation factor IX

Biotechnology Letters - To improve the expression efficiency of recombinant hFIX, by enhancing its γ-carboxylation, which is inhibited by Calumenin (CALU), we used intronic artificial...     

Conformation of a bound inhibitor of blood coagulant factor Xa

13C[(15)N] and (13)C[(19)F] rotational-echo double-resonance NMR have been used to characterize the enzyme-bound structure of ZK-816042, an amidine-imidazoline inhibitor of human factor Xa (FXa). The NMR experiments were performed on a lyophilized FXa-inhibitor complex. The complex was formed in solution in the presence of stabilizing excipients and frozen after gradual supercooling prior to lyophilization. The results indicate that the inhibitor binds with a distribution of orientations of the imidazoline ring.     

Purification and characterization of a coagulant enzyme from Trimeresurus flavoviridis venom

An enzyme bearing thrombin-like specificity has been purified to homogeneity from the venom of Trimeresurus flavoviridis (the Habu snake). The enzyme is a monomer with a molecular weight of 23,500 as determined by analytical gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis. The protein contains approximately 210 amino acid residues and has a relatively high content of aspartic acid and glutamic acid. The isoelectric point was 4.8 and the extinction coefficient at 280 nm for a 1% solution was 11.5. The enzyme acted directly on fibrinogen to form a fibrin clot with 2.0 NIH units. Analysis by high performance liquid chromatography of enzyme-treated fibrinogen revealed the release of a peptide identical in composition to thrombin-induced fibrinopeptide A, but no peptide corresponding to fibrinopeptide B was detected. The enzyme showed esterase and amidase activities on synthetic substrates containing arginine. The enzyme exhibited higher activity toward tosyl-L-arginine methyl ester (TAME) but 6-times lower activity toward benzoyl-L-arginine p-nitroanilide when compared with bovin thrombin. The esterase activity was inhibited by diisopropylfluorophosphate and at a slower rate by phenylmethanesulfonyl fluoride, but was least affected by tosyl-L-lysine chloromethyl ketone, showing that the enzyme is a serine protease like thrombin. The enzyme showed a bell-shaped pH dependence of kcat/Km for hydrolysis of TAME, with a maximum around pH 8.5.     

Structure-activity relationship of a hemagglutinin from Moringa oleifera seeds

The hemagglutinin from the seeds of Moringa oleifera (MoL) agglutinates human as well as rabbit erythrocytes; the affinity for the latter is almost 250 times more than that for the former. MoL was inhibited by glycoproteins namely thyroglobulin, fetuin and holotransferin indicating the complex sugar specificity of the lectin. The protein is a homodimer with molecular mass of 14kDa, subunits (7.1kDa) linked by the disulfide bond(s). The secondary structure elements of MoL are alpha-helix, 28%; beta-sheet, 23%; turn 20% and unordered 28%. While the activity and secondary structure were not affected at extreme pH and high temperature, they were drastically affected in presence of dithiothreitol at and above pH 7.0, indicating that disulfide linkages hold the active conformation of the protein.     

Moringa oleifera oil: a possible source of biodiesel

Biodiesel is an alternative to petroleum-based conventional diesel fuel and is defined as the mono-alkyl esters of vegetable oils and animal fats. Biodiesel has been prepared from numerous vegetable oils, such as canola (rapeseed), cottonseed, palm, peanut, soybean and sunflower oils as well as a variety of less common oils. In this work, Moringa oleifera oil is evaluated for the first time as potential feedstock for biodiesel. After acid pre-treatment to reduce the acid value of the M. oleifera oil, biodiesel was obtained by a standard transesterification procedure with methanol and an alkali catalyst at 60 degrees C and alcohol/oil ratio of 6:1. M. oleifera oil has a high content of oleic acid (>70%) with saturated fatty acids comprising most of the remaining fatty acid profile. As a result, the methyl esters (biodiesel) obtained from this oil exhibit a high cetane number of approximately 67, one of the highest found for a biodiesel fuel. Other fuel properties of biodiesel derived from M. oleifera such as cloud point, kinematic viscosity and oxidative stability were also determined and are discussed in light of biodiesel standards such as ASTM D6751 and EN 14214. The (1)H NMR spectrum of M. oleifera methyl esters is reported. Overall, M. oleifera oil appears to be an acceptable feedstock for biodiesel.