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NASA NTRS2004

Enzymatic Properties of an Alkaline and Chelator Resistant Proportional to alpha-Amylase from the Alkaliphilic Bacillus sp. Isolate L1711

An alkaliphilic amylase producing bacterium, Bacillus sp. strain L1711, was selected among 13 soda lakes isolates. When grown at pH 10.5 and 370 C, strain L1711 produced multiple forms of amylases in the culture broth. One of these, BAA, was purified from the culture supernatant by QAE column chromatography and preparative native gel electrophoresis. The molecular weight of BAA was determined to be 51 kDa by denaturing gel electrophoresis. The pH optima for activity below and above 40 C were 9.5-10.0 and 7.0-7.5 respectively. BAA was stable in the pH range 6-11 and was completely inactivated at 55?C. The thermostability was not increased in the presence of Ca(2+). The enzyme was strongly inhibited by Ca(2+), Zn(2+), Mg(2+), Mn(2+), Ba(2+) and Cu(2+), whereas the presence of Na(+), Co2+ and EDTA (10 mM) enhanced enzymatic activity. The K(sub m) and specific activity of BAA on soluble starch were 1.9 mg/ml and 18.5 U/mg respectively. The main end products of hydrolysis were maltotetraose, maltose and glucose .

Bernhardsdotter, Eva C. M. J.↗

NASA NTRS2001

Beneficial effects of humic acid on micronutrient availability to wheat

Humic acid (HA) is a relatively stable product of organic matter decomposition and thus accumulates in environmental systems. Humic acid might benefit plant growth by chelating unavailable nutrients and buffering pH. We examined the effect of HA on growth and micronutrient uptake in wheat (Triticum aestivum L.) grown hydroponically. Four root-zone treatments were compared: (i) 25 micromoles synthetic chelate N-(4-hydroxyethyl)ethylenediaminetriacetic acid (C10H18N2O7) (HEDTA at 0.25 mM C); (ii) 25 micromoles synthetic chelate with 4-morpholineethanesulfonic acid (C6H13N4S) (MES at 5 mM C) pH buffer; (iii) HA at 1 mM C without synthetic chelate or buffer; and (iv) no synthetic chelate or buffer. Ample inorganic Fe (35 micromoles Fe3+) was supplied in all treatments. There was no statistically significant difference in total biomass or seed yield among treatments, but HA was effective at ameliorating the leaf interveinal chlorosis that occurred during early growth of the nonchelated treatment. Leaf-tissue Cu and Zn concentrations were lower in the HEDTA treatment relative to no chelate (NC), indicating HEDTA strongly complexed these nutrients, thus reducing their free ion activities and hence, bioavailability. Humic acid did not complex Zn as strongly and chemical equilibrium modeling supported these results. Titration tests indicated that HA was not an effective pH buffer at 1 mM C, and higher levels resulted in HA-Ca and HA-Mg flocculation in the nutrient solution.

Non-NASA Center↗

NASA NTRS1989

Calcium bridges are not load-bearing cell-wall bonds in Avena coleoptiles

I examined the ability of frozen-thawed Avena sativa L. coleoptile sections under applied load to extend in response to the calcium chelators ethyleneglycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) and 2-[(2-bis-[carboxymethyl]amino-5-methylphenoxy)methyl]-6-methoxy-8-bis[car boxymethyl]aminoquinoline (Quin II). Addition of 5 mM EGTA to weakly buffered (0.1 mM, pH 6.2) solutions of 2(N-morpholino) ethanesulfonic acid (Mes) initiated rapid extension and wall acidification. When the buffer strength was increased (e.g. from 20 to 100 mM Mes, pH 6.2) EGTA did not initiate extension nor did it cause wall acidification. At 5 mM Quin II failed to stimulate cell extension or wall acidification at all buffer molarities tested (0.1 to 100 mM Mes). Both chelators rapidly and effectively removed Ca2+ from Avena sections. These data indicate that Ca2+ chelation per se does not result in loosening of Avena cells walls. Rather, EGTA promotes wall extension indirectly via wall acidification.

NASA Program Space Biology↗

NASA NTRS1985

Role of Ca++ in Shoot Gravitropism

A cornerstone in the argument that Ca(2+) levels may regulate growth is the finding the EGTA promotes straight growth. The usual explanation for these results is that Ca(2+) chelation from cell walls results in wall loosening and thus accelerated straight growth. The ability of frozen-thawed Avena coleoptile tissue (subjected to 15g tension) to extend in response to EGTA and Quin II was examined. The EGTA when applied in weakly buffered (i.e., 0.1mM) neutral solutions initiates rapid extension. When the buffer strength is increased, similar concentrations of EGTA produce no growth response. This implies when EGTA liberated protons are released upon Ca(2+) chelation they can either initiate acid growth (low buffer conditions) or if consumed (high buffer conditions) have no effect. Thus Ca(2+) chelation in itself apparently does not result in straight growth.

Rayle, D. L.↗

NASA NTRS1998

Calcium-independent activation of extracellular signal-regulated kinases 1 and 2 by cyclic strain

We have previously demonstrated that cyclic strain induces extracellular signal-regulated kinases 1 and 2 (ERK1/2) activation in endothelial cells (EC). The aim of this study was to investigate the effect of Ca2+ on the activation of ERK1/2. Bovine aortic EC were pretreated with a chelator of extracellular Ca2+, ethylaneglycol-bis(aminoethylether)-tetra-acetate (EGTA), a depleter of Ca2+ pools, 2,5-Di-(tert-butyl)-1,4-benzohydroquinone (BHQ), or a Ca2+ channel blocker, GdCl3, and subjected to an average 10 % strain at a rate of 60 cycles/min for 10 min. BHQ and GdCl3 did not inhibit the strain-induced ERK1/2 activation. Chelation of normal extracellular Ca2+ (1.8 mM) medium with EGTA (3 mM) acutely stimulated baseline phosphorylation and activation of ERK1/2, thereby obscuring any strain-induced activation of ERK1/2. However, in EC preincubated for 24 hours in Ca2+-free medium, elevated baseline phosphorylation was minimally activated by EGTA (200 microM) such that cyclic strain stimulated ERK1/2 in the presence or absence of BHQ. These results suggest a Ca2+ independence of the ERK1/2 signaling pathway by cyclic strain. Copyright 1998 Academic Press.

NASA Discipline Cell Biology↗

NASA NTRS1969

Liquid laser cavities

Liquid laser cavities have plenum chambers at the ends of the capillary cell which are terminated in transparent optical flats. By use of these cavities, several new europium chelates and a terbium chelate can provide laser action in solution at room temperature.

Bjorklund, S.↗

NASA NTRS1972

Advanced techniques for determining long term compatibility of materials with propellants

The search for advanced measurement techniques for determining long term compatibility of materials with propellants was conducted in several parts. A comprehensive survey of the existing measurement and testing technology for determining material-propellant interactions was performed. Selections were made from those existing techniques which were determined could meet or be made to meet the requirements. Areas of refinement or changes were recommended for improvement of others. Investigations were also performed to determine the feasibility and advantages of developing and using new techniques to achieve significant improvements over existing ones. The most interesting demonstration was that of the new technique, the volatile metal chelate analysis. Rivaling the neutron activation analysis in terms of sensitivity and specificity, the volatile metal chelate technique was fully demonstrated.

Green, R. L.↗

NASA NTRS1981

Development of W-Ta generator

This research program was used to further develop the existing W-Ta generator and to evaluate alternative adsorbents, preferably inorganic materials, as supports for the generator. During the first half year, combinations of non-complexing eluents and a variety of adsorbents, both inorganic and organic, were evaluated. Some of these adsorbents were synthetic, such as chelate resins that could be specific for tungsten. In the second half of the year, the stress was mainly on the use of complexing eluents because of the high affinity of hydrous oxides for tantalum, on the synthesis of chelate resins and on the use novel techniques (electrolytic) to solve the tantalum-adsorption problem.

Source record↗

NASA NTRS1985

Effects of method of detachment on electrophoretic mobility of mammalian cells grown in monolayer culture

A variety of proteolytic and micolytic enzumes, mechanical procedures, and changes in the ionic environment, especially Ca chelation, are used for dispersal of monolayer grown cells. If either chelating agents or mechanical dispersion are used alone, the cell yield is often low and suspensions of single cells are difficult to obtain. Confluent monolayers treated with EDTA tend to be released from their surfaces in sheets, and clumps of cells remain even after further incubation in EDTA. Crude trypsin is the most popular dispersal agent and is known to contain a variety of contaminating enzymes which contribute to the dispersal of cells. A variety of cell injuries resulting from the activity of proteolytic enzymes are reported. It is shown that crystalline trypsin is least harmful to cell integrity as judged by trypan blue uptake.

Plank, L. D.↗

NASA NTRS1990

Recovery of precious metals in space

This program is proceeding along the lines compatible with the objective of developing feasible separation and recovery processes for precious metals in a space environment. Efficient, compact, solvent extraction systems (both single and multistage) capable of high selectivity and rapid equilibration that are based on chelate and ion pair complexes are being explored. Two research projects were undertaken: (1) study of the time-dependent aspects of the formation of Pd(2) extractable chelate complexes; and (2) development of a multistage countercurrent distribution process for the separation and recovery of Pd(2) and Pt(2) ions.

Freiser, H.↗

NASA NTRS1993

Determination of trace elements in triglycine sulfate solutions

Ten elements were divided into 2 groups. The elements in the first group included iron, nickel, chromium, manganese, copper, and gold. The elements in the second group included zinc, cobalt, lead, cadmium, and gold. Five ppm of each element in each group was spiked in a 1 percent triglycine sulfate (TGS) solution. Glycine was removed with 1-naphthyl isocyanate in ether medium. The glycine derivative 1-naphthyl isocyanate glycine was removed by filtration, and the filtrates were analyzed for the different elements. Analysis of these elements was performed by using the 5100 Perkin-Elmer Atomic Absorption Spectrophotometer. The result of these experiments was the observation that there was a decrease in the concentration of chromium and gold, which was interpreted to be due to the chelation of these elements by the derivative 1-naphthyl isocyanate glycine. Further research is needed to determine the concentration of other elements in triglycine sulfate (TGS) solutions. These elements will include lithium, sodium, rubidium, magnesium, calcium, strontium, barium, aluminum, and silicon. These are the most likely elements to be found in the sulfuric acid used in manufacturing the TGS crystal. Moreover, we will extend our research to investigate the structural formula of the violet colored chelated compounds, which had been formed by interaction of the derivative 1-naphthyl isocyanate glycine with the different elements, such as gold, chromium.

Tadros, Shawky H.↗

NASA NTRS2003

Biogeochemistry of fluoride in a plant-solution system

Fluoride (F-) pollutants can harm plants and the animals feeding on them. However, it is largely unknown how complexing and chelating agents affect F bioavailability. Two studies were conducted that measured F- bioavailability and uptake by rice (Oryza sativa L.). In the first study, rice was grown in solution culture (pH 5.0) with 0, 2, or 4 mM F- as KF to compare the interaction of F- with humic acid (HA) and with a conventional chelating agent, N-hydroxyethylenthylenediaminetriacetic acid (HEDTA). In the second study, F was supplied at 0, 0.5, 1.0, and 2.0 mM KF with an additional 2 mM F- treatment containing solution Ca at 2x (2 mM Ca) the level used in the first study, to test the effect added Ca had on F- availability and uptake. Total biomass was greatest with HEDTA and F- < 1 mM. Leaf and stem F concentrations increased exponentially as solution F- increased linearly, with nearly no F partitioning into the seed. Results suggest that F was taken up as HF0 while F- uptake was likely restricted. Additionally, F- competed with HA for Ca, thus preventing the formation of Ca-HA flocculents. The addition of soluble Ca resulted in the precipitation of CaF2 solids on the root surface, as determined by tissue analysis and energy dispersive X-ray spectroscopy.

NASA Discipline Life Support Systems↗

NASA NTRS1981

Functional magnetic microspheres

Functional magnetic particles are formed by dissolving a mucopolysaccharide such as chitosan in acidified aqueous solution containing a mixture of ferrous chloride and ferric chloride. As the pH of the solution is raised magnetite is formed in situ in the solution by raising the pH. The dissolved chitosan is a polyelectrolyte and forms micelles surrounding the granules at pH of 8-9. The chitosan precipitates on the granules to form microspheres containing the magnetic granules. On addition of the microspheres to waste aqueous streams containing dissolved ions, the hydroxyl and amine functionality of the chitosan forms chelates binding heavy metal cations such as lead, copper, and mercury and the chelates in turn bind anions such as nitrate, fluoride, phosphate and borate.

Yen, Shiao-Ping S.↗

NASA NTRS1985

A technique for collection of exudate from pea seedlings

Ethylenediaminetetraacetic acid (EDTA), at concentrations higher than 1.0 millimolar, is phytotoxic to etiolated seedlings of Pisum sativum. Substantial vascular exudation from pea epicotyls could be obtained without tissue damage at 0.5 millimolar EDTA if the solution was buffered at pH 7.5 with sodium N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid. Treated seedlings exuded 950 micrograms (leucine equivalents) of ninhydrin-positive material per day and 870 micrograms (glucose equivalents) of anthrone-positive material per day. Amino acid analysis showed the exudate to have glutamine as the major amido nitrogen containing compound and sucrose was shown to be the major sugar. Radiolabeled tryptophan and sucrose applied to cotyledons were transferred through the epicotyl and into the collection medium. The pH profile for exudation shows half maximal exudation at pH 7.2, indicating the promotion of exudation by EDTA is probably not due simply to Ca2+ chelation.

Non-NASA Center↗

NASA NTRS1988

Effects of cations on hormone transport in primary roots of Zea mays

We examined the influence of aluminum and calcium (and certain other cations) on hormone transport in corn roots. When aluminum was applied unilaterally to the caps of 15 mm apical root sections the roots curved strongly away from the aluminum. When aluminum was applied unilaterally to the cap and 3H-indole-3-acetic acid was applied to the basal cut surface twice as much radioactivity (assumed to be IAA) accumulated on the concave side of the curved root as on the convex side. Auxin transport in the apical region of intact roots was preferentially basipetal, with a polarity (basipetal transport divided by acropetal transport) of 6.3. In decapped 5 mm apical root segments, auxin transport was acropetally polar (polarity = 0.63). Application of aluminum to the root cap strongly promoted acropetal transport of auxin reducing polarity from 6.3 to 2.1. Application of calcium to the root cap enhanced basipetal movement of auxin, increasing polarity from 6.3 to 7.6. Application of the calcium chelator, ethylene-glycol-bis-(beta-aminoethylether)-N,N,N',N'-tetraacetic acid, greatly decreased basipetal auxin movement, reducing polarity from 6.3 to 3.7. Transport of label after application of tritiated abscisic acid showed no polarity and was not affected by calcium or aluminum. The results indicate that the root cap is particularly important in maintaining basipetal polarity of auxin transport in primary roots of corn. The induction of root curvature by unilateral application of aluminum or calcium to root caps is likely to result from localized effects of these ions on auxin transport. The findings are discussed relative to the possible role of calcium redistribution in the gravitropic curvature of roots and the possibility of calmodulin involvement in the action of calcium and aluminum on auxin transport.

NASA Discipline Plant Biology↗

NASA NTRS1988

Calcium in the regulation of gravitropism by light

The red light requirement for positive gravitropism in roots of corn (Zea mays cv "Merit") provides an entry for examining the participation of calcium in gravitropism. Applications of calcium chelators inhibit the light response. Calcium channel blockers (verapamil, lanthanum) can also inhibit the light response, and a calcium ionophore, A23187, can substitute for light. One can substitute for red light by treatments which have elsewhere been shown to trigger Ca2+ influx into the cytosol, e.g. heat or cold shock. Agents which are known to be agonists of the phosphatidylinositol second messenger system (serotonin, 2,4-dichlorophenoxyacetic acid, deoxycholate) can each partially substitute for the red light, and Li+ can inhibit the light effect. These experiments suggest that the induction of positive gravitropism by red light involves a rise in cytoplasmic Ca2+ concentration, and that a contribution to this end may be made by the phosphatidylinositol second messenger system.

NASA Discipline Number 40-10↗

NASA NTRS1997

Activation of G proteins mediates flow-induced prostaglandin E2 production in osteoblasts

Interstitial fluid flow may play a role in load-induced bone remodeling. Previously, we have shown that fluid flow stimulates osteoblast production of cAMP inositol trisphosphate (IP3), and PGE2. Flow-induced increases in cAMP and IP3 were shown to be a result of PG production. Thus, PGE2 production appears to be an important component in fluid flow induced signal transduction. In the present study, we investigated the mechanism of flow-induced PGE2 synthesis. Flow-induced a 20-fold increase in PGE2 production in osteoblasts. Increases were also observed with ALF4-(10mM) (98-fold), an activator of guanidine nucleotide-binding proteins (G proteins), and calcium ionophore A23187 (2 microM) (100-fold) in stationary cells. We then investigated whether flow stimulation is mediated by G proteins and increases in intracellular calcium. Flow-induced PGE2 production was inhibited by the G protein inhibitors GDP beta S (100 microM) and pertussis toxin (1 microgram/ml) by 83% and 72%, respectively. Chelation of extracellular calcium by EGTA (2 mM) and intracellular calcium by quin-2/AM (30 microM) blocked flow stimulation by 87% and 67%, respectively. These results suggest that G proteins and calcium play an important role in mediating mechanochemical signal transduction in osteoblasts.

NASA Discipline Cell Biology↗

NASA NTRS1961

Kinetics of the Reaction Between Alcohols and Isocyanates Catalyzed by Ferric Acetylacetonate

The rate and temperature dependence of reaction for the ferric acetylacetonate catalyzed reaction between a-naphthyl, ortho-tolyl, and para-tolyl isocyanates and n-butyl alcohol are investigated. The effect of substituents on the reactivity of isocyanate and hydroxyl group are reported and for substituted isocyanates are correlated by means of the Hammett equation. Several metal chelates were studied and their catalytic activity was compared to that of ferric acetylacetonate. All rate data are interpreted in terms of a mechanism involving simultaneous second-order uncatalyzed and catalyzed reactions between alcohol and isocyanate.

ACETONE↗