Calcium transport by pea root membranes

Calcium transport has been studied using purified endomembrane vesicles from dark-grown roots of Pisum sativum L. Membranes from a mixed microsomal (non-mitochondrial) fraction showed ATP-dependent calcium uptake which was released by the ionophore A 23187, had a pH optimum of 7.2 and required Mg²⁺ for uptake. Membranes were further purified using a rapid sucrosedensity-gradient technique yielding vesicles suitable for transport studies, and were identified using marker enzymes. Uptake by plasma membrane, tonoplast, endoplasmic reticulum and Golgi apparatus was indicated. Uptake by membranes of low density (predominantly tonoplast) had a pH optimum of 7.2–7.4 and nucleotide specificity ATP> guanosine 5-triphosphate>inosine 5-triphosphate>ADP>, while that by high-density membranes had a pH optimum of 7.5–7.9 and less specificity for ATP. The importance of regulating sucrose concentrations in calcium transport studies was demonstrated.Abbreviations ER endoplasmic reticulum - GTP guanosine 5-triphosphate - IDPase inosine diphosphatase - IIP inosine 5-triphosphate     

Year-class strength regulation in plaice (Pleuronectes platessa L.) on the Swedish west coast

Year-class strength of plaice (Pleuronectes platessa) has been studied over periods of 10 and 35 years, respectively, in two shallow sandy areas on the Swedish west coast. In one area, Gullmar Bay (N 58 ° 19 – W 11 ° 33), 0-group plaice were sampled quantitatively with a drop trap at 0–0.7 m depth and densities between 0.2 and 3.8 ind./m² were recorded in early summer. In the other area, Laholm Bay (N 56 ° 30 – W 12 ° 55), 0-group plaice were sampled semi-quantitatively in August with young-fish trawl at 1.5 m depths. Densities between 0.001 and 0.28 ind./m² were assessed.The effects of temperature and wind (in the winter and early spring) and predation (in early summer), on the recruitment of 0-group plaice were investigated. In Gullmar Bay high recruitment occurred after severe winters and in years when on-shore winds dominated during spring. No such correlations were found in Laholm Bay. In Gullmar Bay a significant inverse relationship was found between the density of O-group plaice in early summer and the biomass of brown shrimp (Crangon crangon) and the shore crab (Carcinus maenas), the main predators in the nursery area.The importance of physical and biological factors regulating recruitment in plaice are discussed.     

Observation of a distinct transition in the mode of interconversion of ring pucker conformers in non-crystalline d-ribose-2'-d from 2H NMR spin-alignment

Internal motions of d-ribose selectively ²H-labeled at the 2 position were measured using solid state ²H NMR experiments. A sample of d-ribose-2 -d was prepared in a hydrated, non-crystalline state to eliminate effects of crystal-packing. Between temperatures of –74 and –60°C the C2–H2 bond was observed to undergo two kinds of motions which were similar to those of C2–H2/H2 found previously in crystalline deoxythymidine (Hiyama et al. (1989) J. Am. Chem. Soc., 111, 8609–8613): (1) Nanosecond motion of small angular displacement with an apparent activation energy of 3.6 ± 0.7 kcal mol^–1, and (2) millisecond to microsecond motion of large amplitude with an apparent activation energy 4 kcal mol^–1. At –74°C, the slow, large-amplitude motion was best characterized as a two-site jump with a correlation time on the millisecond time scale, whereas at –60°C it was diffusive on the microsecond time scale. The slow, large-amplitude motions of the C2–H2 bond are most likely from interconversions between C2-endo and C3-endo by way of the O4-endo conformation, whereas the fast, small-amplitude motions are probably librations of the C2–H2 bond within the C2-endo and C3-endo potential energy minima.     

Yearly reproductive activity of Limnoperna fortunei (Bivalvia) as inferred from the occurrence of its larvae in the plankton of the lower Paraná river and the Río de la Plata estuary (Argentina)

Densities of planktonic larvae of the Asian freshwater invasive pest bivalve Limnoperna fortunei (Dunker) were monitored weekly between 19 September 1997 and 22 January 1999 at two sites: the Paraná de las Palmas river, ca. 130 km north of Buenos Aires (33°57.5 S, 59°12.5 W), and a coastal station on the Río de la Plata estuary, in Buenos Aires (34°33.5 S, 58°24.5 W). Very variable but generally high (up to 25000–35000 ind. m^–3) concentrations were recorded at both sites in September 1997-April 1998, and again in August 1998–January 1999, suggesting continuous reproduction during these months. In May through July 1998 larval densities were generally below 100-200 ind. m^–3. The threshold temperature level associated with the onset and termination of reproduction was ca. 16–17 °C. With the exception of August-September 1998, when Río de la Plata temperatures were below 16.7 °C and larval concentrations less than 1000 ind. m^–3, whereas those of the Paraná were above 16.7 °C, with 1300–13000 larvae m^–3, coupling between the two sites investigated was generally good. Since its invasion in Argentina around 1990, L. fortunei has rapidly become a major nuisance for industrial and power plants located along the lower Paraná and Río de la Plata. The fact that the mollusc produces larvae during 9 months per year greatly hinders its control.     

Modelling of temperature effects on batch microbial transglutaminase fermentation with Streptoverticillium mobaraense

Batch transglutaminase (MTG) fermentations by Streptovertivillium mobaraense WSH-Z2 at various temperatures ranging from 25 to 35 °C were studied. Dry cell weight and MTG activity could reach their maximal values of 25.1 g/l and 2.94 U/ml, respectively at 30 °C. One typical equation was used to describe the relationship between specific growth rate and culture temperature by comparing several typical equations. Different lag time was observed under various culture temperature. The low lag time was observed under high culture temperature. X = –a ₀(T – T ₀)² + X ₁ + a ₁ (1 – exp(a ₂ (T – T ₁))) and U = –a ₀ (T – T ₀ )² + U ₀ + a ₁ (1 – exp(a ₂ (T – T ₁ ))) could be used to describe the relationship between temperature and the maximal dry cell weight as well as the maximal MTG activity at each temperature.     

Na/H exchange in cultured epithelial cells from fish gills

Using primary cultures of gill pavement cells from freshwater rainbow trout, a method is described for achieving confluent monolayers of the cells on glass coverslips. A continuous record of intracellular pH was obtained by loading the cells with the pH-sensitive flourescent dye 2,7-bis(2-carboxyethyl)-5(6)-carboxyfluorescein and mounting the coverslips in the flowthrough cuvette of a spectrofluorimeter. Experiments were performed in HEPES-buffered media nominally free of HCO₃. Resting intracellular pH (7.43 at extracellular pH=7.70) was insensitive to the removal of Cl^– or the application of 4-acetamido-4-isothiocyanatostilbene-2,2-disulfonic acid (0.1 mmol·l^–1), but fell by about 0.3 units when Na⁺ was removed or in the presence of amiloride (0.2 mmol·l^–1). Exposure to elevated ammonia (ammonia prepulse; 30 mmol·l^–1 as NH₄Cl for 6–9 min) produced an increase in intracellular pH (to about 8.1) followed by a slow decay, and washout of the pulse caused intracellular pH to fall to about 6.5. Intracellular non-HCO ₃ ^– buffer capacity was about 13.4 slykes. Rapid recovery of intracellular pH from intracellular acidosis induced by ammonia prepulse was inhibited more than 80% in Na⁺-free conditions or in the presence of amiloride (0.2 mmol·l^–1). Neither bafilomycin A₁ (3 mol·l^–1) nor Cl removal altered the intracellular pH recovery rate. The K _m for Na⁺ of the intracellular pH recovery mechanism was 8.3 mmol·l^–1, and the rate constant at V _max was 0.008·s^–1 (equivalent to 5.60 mmol H⁺·l^–1 cell water·min^–1), which was achieved at external Na⁺ levels from 25 to 140 mmol·l^–1. We conclude that intracellular pH in cultured gill pavement cells in HEPES-buffered, HCO ₃ ^– -free media, both at rest and during acidosis, is regulated by a Na⁺/H⁺ antiport and not by anion-dependent mechanisms or a vacuolar H⁺-ATPase.Abbreviations BCECF 2,7-bis(2-carboxyethyl)-5(6)-carboxy-fluorescein - BCECF/AM 2,7-bis(2-carboxyethyl)-5(6)-carboxy-fluorescein, acetoxymethylester - Cholin-Cl choline chloride - DMSO dimethyl sulfoxide - EDTA ethylene diamine tetra-acetic acid - FBS foetal bovine serum - H ⁺ -ATPase Proton-dependent adenosine triphosphatase - HEPES N-[2-hydroxyethyl]piperazine-N[2-ethanesulfonic acid] - pH _i intracellular pH - pH _e extracellular pH - PBS phosphate-buffered saline - SITS 4-acetamido-4-isothiocyanatostilbene-2,2-disulfonic acid     

Oligonucleotide formation catalyzed by divalent metal ions. The uniqueness of the ribosyl system

Summary Polymerization of various nucleoside-5-phosphorimidazolides has been conducted in neutral aqueous solution using divalent metal ions as catalysts. Oligonucleotide formation took place from each of the ribonucleoside-5-phosphorimidazolides, ImpC, ImpU, ImpA, ImpG, and ImpI. The yields and distributions of the resulting oligonucleotides varied depending on the difference of the nucleic acid base and the metal ions used. The catalytic effect of divalent metal ions on the formation of oligocytidylates occurred in the following order: Pb²⁺>Zn²⁺>Co²⁺, Mn²⁺>Cd²⁺>Cu²⁺>Ni²⁺>Ca²⁺, Mg²⁺, none >Hg²⁺. The order changes slightly for other types of oligoribonucleotide formation. Oligoribonucleotides up to hexamers were obtained in 35–55% overall yield, when Pb²⁺ ion was used as a catalyst. Zn²⁺ ions yielded oligoribonucleotides up to tetramers in 10–20% overall yield. The resulting oligonucleotides contained mainly 2–5 internucleotide linkages.Little or no oligonucleotide was obtained from nucleoside-5-phosphorimidazolides modified in the sugars, Imp(3-dA), Imp(2-dA), Imp(Ara), Imp(Aris), and Imp(Nep). The results indicate that a ribosyl system is required for the metal ion-catalyzed synthesis of oligonucleotides. Abbreviations. EDTA, ethylenediaminetetraacetic acid; Versenol,N-hydroxyethylethylenediaminetriacetic acid; Tris, tris-(hydroxymethyl)aminomethane; pN (N is A, C, G, U, I, 3-dA, 2-dA, AraA, Aris, or Nep), nucleoside-5-phosphate; Np, nucleoside-2(3)-phosphate; I, inosine; 3-dA, 3-deoxyadenosine; 2-dA, 2-deoxyadenosine; AraA, arabinosyladenine; Aris, aristeromycin; Nep, neplanocin A; ImpN, nucleoside-5-phosphorimidazolide; NppN, P₁,P₂-dinucleoside-5,5-pyrophosphate; (pN)_n (n=2, 3, ...), oligomers of pN, numbers given between a nucleoside and a phosphate indicate the type of internucleotide linkage, e.g., pC₂ p₅C is 5-phosphorylcytidyl-(2–5)-cytidine; , cyclic dimers of pN; BAP, bacterial alkaline phosphatase; N.Pl, nuclease Pl; VPDase, venom phosphodiesterase; HPLC, high pressure liquid chromatography     

Surface phytoplankton composition and carbon biomass distribution in the Crozet Basin during austral summer of 1999: variability across frontal zones

Surface phytoplankton assemblages were studied in January/February 1999 in the Crozet Basin (43°50S–45°20S; 61°E–64°30E) between the northern Polar Zone and the Agulhas Front. Cell concentrations increased several fold northwards from the SubAntarctic Zone (SAZ) and reached peak numbers (average 2×10⁶ cells l^–l ) in the central and western Subtropical Zone (STZ). The most spectacular increase in cell numbers occurred at the Subtropical Front (STF) and was attributed to dinoflagellates and diatoms. Nanoflagellates and picoplankton were dominant in the entire area (average 2.8×10⁵–1.6×10⁶ cells l^–l). In the SAZ they were followed by coccolithophorids, dinoflagellates and diatoms. In the STZ coccolithophorids were often outnumbered by dinoflagellates. Diatoms were dominated by Pseudonitzschia delicatissima and were generally the least abundant algae, but reached peak densities of 1.2–4×10⁵ cells l^–l at, and north of the STF. Coccolithophorids contained mainly Emiliania huxleyi, but in the SAZ and STF Gephyrocapsa oceanica was a co-dominant species. Dinoflagellates were dominated by nano-sized species of Gymnodinium, Gyrodinium and Prorocentrum. The numbers of dinoflagellate and coccolithophorid species increased considerably in the convergence zone (STZ), which suggests their in-situ development. Heterotrophic dinoflagellates and ciliates were mainly present in the subtropics. Cell carbon biomass was attributed chiefly to auto- and heterotrophic dinoflagellates (av. 23–72 g C l^–l; 68–87%), showing their important contribution to the carbon flow. Variations in cell concentrations across the fronts and water masses, and the distribution of major species were most likely controlled by the combined effect of such factors as nutrient renewal in the convergence zone, availability of iron, increased water-column stability at fronts, and high horizontal gradients in surface-water temperature.     

pH equilibration in human erythrocyte suspensions

Summary A stopped-flow rapid reaction apparatus was used to study the rate of pH equilibration in human red cell suspensions. Flux of OH^– or H⁺ was determined over a wide range of extracellular pH (4–11) in CO₂-free erythrocyte suspensions. In these experiments, an erythrocyte suspension at pH 7.3 is rapidly mixed with an equal volume of NaCl solution at 3.0>pH>11.5. The pH of the extracellular fluid of the mixture changes rapidly as OH^– or H⁺ moves across the red cell membrane. Flux and velocity constants can be calculated from the initiald pH/dt using the known initial intra- and extracellular pH. It was found that the further the extracellular pH is from 7.3 (in either direction from 4–11), the greater the absolute value of total OH^– and/or H⁺ flux. Pretreatment with SITS (4-acetamido-4-isothiocyanostilbene-2,2-disulfonic acid), a potent anion exchange inhibitor, greatly reduces flux over the entire pH range, while exposure to valinomycin, a potassium ionophore, has no measurable effect. These data suggest that (i) both H⁺ and OH^– may be moving across the red cell membrane at all pH; (ii) the species dominating pH equilibration is probably dependent on the extracellular pH, which determines the magnitude of the driving gradient for each ion; and (iii) the rapid exchange pathway of the erythrocyte membrane may be utilized for both H⁺ and OH^– transport during CO₂-free pH equilibration.     

Genetic evidence for the nature, and excision repair, of DNA lesions resulting from incorporation of 5-bromouracil

Summary Escherichia coli mutants defective in DNA uracil N-glycosidase (ung ^–) or endonuclease VI active against apurinic/apyrimidinic sites in DNA (xthA^–) exhibit enhanced sensitivity towards 5-bromodeoxyuridine relative to the wild type strain, pointing to involvement of these enzymes in repair of bromouracil-induced lesions in DNA.Mutants defective in DNA polymerase I, either in polymerizing activity (polAl^–) or (53)-exonuclease activity (polA107^–) exhibit unusually high sensitivity (including marked lethality) in the presence of 5-bromodeoxyuridine. The results indicate that DNA polymerase I, and its associated (5–3)-exonuclease activity, are involved in repair of bromouracil-induced lesions and are not readily replaced, if at all, by DNA polymerases II and III.Thermosensitive mutant in DNA ligase gene (lig ts7) shows high sensitivity towards 5-bromodeoxyuridine at 42°C indicating the role of the enzyme in repair of bromouracil-induced lesions in DNA.Involvement of DNA uracil N-glycosidase, and endonuclease active against apurinic/apyrimidinic sites in recognition and repair of 5-bromouracil-induced damage permits of some inferences regarding the nature of this damage (lesions), in particular dehalogenation of incorporated bromouracil to uracil residues.     

Cell swelling activates separate taurine and chloride channels in Ehrlich mouse ascites tumor cells

The taurine efflux from Ehrlich ascites tumor cells is stimulated by hypotonic cell swelling. The swelling-activated taurine efflux is unaffected by substitution of gluconate for extracellular Cl^– but inhibited by addition of MK196 (anion channel blocker) and 4,4-diisothiocyanostilbene-2,2-disulfonic acid (DIDS; anion channel and anion exchange blocker) and by depolarization of the cell membrane. This is taken to indicate that taurine does not leave the osmotically swollen Ehrlich cells in exchange for extracellular Cl^–, i.e., via the anion exchanger but via a MK196- and DIDS-sensitive channel that is potential dependent. An additional stimulation of the swelling-activated taurine efflux is seen after addition of arachidonic acid and oleic acid. Cell swelling also activates a Mini Cl^– channel. The Cl^– efflux via this Cl^– channel, in contrast to the swelling-activated taurine efflux, is unaffected by DIDS and inhibited by arachidonic acid and oleic acid. It is suggested that the swelling-activated Mini Cl^– channel and the swelling-activated taurine channel in the Ehrlich cell represent two distinct types of channels.This work was supported by the Danish Natural Research Council and by the NOVO foundation. Dr. Birte Kramhøft is acknowledged for critical reading of this paper.     

Age Dependence of the Level ofcopia Retrotransposon Expression in the Testes of Drosophila melanogaster

Expression of the lacZ reporter gene controlled by various deletion derivatives of the regulatory region of the copia retrotransposon was studied in the testes of adult transgenic males of the Drosophila melanogaster y ¹ w ^67c23(2) strain at the age of 3, 6–9, 12–15, 18–21, and 24–27 days. When the construct contained the full-length regulatory region, which included the 5-long terminal repeat (LTR) and the 5-untranslated region (UTR), expression was the lowest in males aged 12–15 days and the highest in males aged 3 or 24–27 days. A similar V-shaped age dependence was previously observed for the copia transposition rate and RNA content in a strain with a high rate of copia transposition. Thus, the V-shaped age dependence of expression, which is unusual for Drosophila, proved to be characteristic of copiaregardless of its transposition rate. Deletion of the 5 or 3 end of the LTR, but not of the UTR, changed the age dependence of the level of reporter gene expression. In this case, expression was the highest in 3-day-old males and gradually decreased with age, as typical for many Drosophila genes. It was assumed that the 5- and 3-terminal regions of the copiaLTR contain regulatory elements responsible for the V-shaped age dependence of expression, while the expression level depends to a greater extent on the regulatory elements of UTR.     

Characteristics of βA chromosome haplotypes in Japanese

DNA polymorphism patterns linked to the ^A-globin gene were analyzed in healthy Japanese using four different restriction endonucleases. The chromosomes with the ^A-globin gene were mapped through an evaluation of the presence of seven different restriction sites (HincII 5 to ; HindIII in ^G and ^A; HincII in, and 3 to, ₁; AvaII in ; Bam-HI 3 to ). Among 36 chromosomes analyzed, 20 chromosomes had a haplotype of [+–––––+]. Among 55 individuals examined, 7 possessed a homozygous haplotye of [+–––––+]. All Japanese with the ^AT-globin gene had a subhaplotype of [–++–+] 5 to the -globin gene. Their major haplotypes were [–++–+–+] and [–++–++–]. It was expected that the presence of the ^AT-globin gene in Japanese may be deduced from subhaplotypes 5 to the -globin gene.     

Purification and characterization of the 5′ → 3′ exonuclease domain-deleted Thermus filiformis DNA polymerase expressed in Escherichia coli

The gene encoding 5 3 exonuclease domain-deleted Tfi DNA polymerase, named 5 3 Exo^– Tfi fragment, from Thermus filiformis was expressed in Escherichia coli under the control of the tac promoter on a high-copy plasmid, pJR. The expressed enzyme was purified 27-fold with a 19% yield and a specific activity of 2621 U mg^–1 protein. The 5 3 exonuclease domain of Tfi DNA polymerase was removed without significant effect on enzyme activity and stability. PCR conditions for the 5 3 Exo^– Tfi fragment were more tolerant to the buffer composition as compared to the full-length enzyme.     

Increase in chloroplastic thiol groups by SO2 and its effect on light modulation of NADP-dependent glyceraldehyde 3-phosphate dehydrogenase

In broken spinach chloroplasts the total amount of thiol groups is about 3.7 mol mg^-1 chlorophyll. Two thirds are represented by the masked form (which is only titratable after unfolding of the protein). Of the free groups, those reacting with NBD·Cl (1.2–2.0 mol mg^-1 chlorophyll) seem to be undergoing oxidation more readily than those reacting with DTNB (1.0 mol mg^-1 chlorophyll). SO₂ application causes a maximal increase of 25% in free thiols, and doubles the amount of the masked thiols. The light triggered increase in SH, which starts at an elevated level, runs parallel to that of the controls. SO₂ application of 1.8 mg m^-3 (=28 nmol l^-1) for 1 h does not affect the dark level of NADP-GPD but enhances the light modulation by increasing the ratio of activation. This enhancement is explained by an increase in masked thiol groups during the preceding fumigation period.Abbreviations DTNB 5,5 dithiobis-2-nitrobenzene-2-oxa-1,3 diazole - NBD·Cl 7-chloro-4-nitrobenzene-2-oxa-1,3 diazole - PCMB p-chloromercuribenzoate - SDS sodium dodecylsulfate - NADP-GPD NADP-dependent glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.13) - HEPES N-2-Hydroxyethylpiperazine-N-2-ethanesulfonic acid - MES 2[N-Morpholino]ethanesulfonic acid - PGA 3-phosphoglyceric acid     

Functional analysis of the iron-stress induced CP 43′ polypeptide of PS II in the cyanobacterium Synechococcus sp. PCC 7942

Under conditions of iron-stress, the Photosystem II associated chlorophyll a protein complex designated CP 43, which is encoded by the isiA gene, becomes the major pigment-protein complex in Synechococcus sp. PCC 7942. The isiB gene, which is located immediately downstream of isiA, encodes the protein flavodoxin, which can functionally replace ferredoxin under conditions of iron stress. We have constructed two cyanobacterial insertion mutants which are lacking (i) the CP 43 apoprotein (designated isiA ^–) and (ii) flavodoxin (designated isiB ^–). The function of CP 43 was studied by comparing the cell characteristics, PS II functional absorption cross-sections and Chl a fluorescence parameters from the wild-type, isiA ^– and isiB ^– strains grown under iron-stressed conditions. In all strains grown under iron deprivation, the cell number doubling time was maintained despite marked changes in pigment composition and other cell characteristics. This indicates that iron-starved cells remained viable and that their altered phenotype suggests an adequate acclimation to low iron even in absence of CP 43 and/or flavodoxin. Under both iron conditions, no differences were detected between the three strains in the functional absorption crossection of PS II determined from single turnover flash saturation curves of Chl a fluorescence. This demonstrates that CP 43 is not part of the functional light-harvesting antenna for PS II. In the wild-type and the isiB ^– strain grown under iron-deficient conditions, CP 43 was present in the thylakoid membrane as an uncoupled Chl-protein complex. This was indicated by (1) an increase of the yield of prompt Chl a fluorescence (F_o) and (2) the persistence after PS II trap closure of a fast fluorescence decay component showing a maximum at 685 nm.Abbreviations Chl chlorophyll - CP 43, CP 47 and CP 43 Chl a binding protein complexes of indicated molecular mass - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_m and F_m fluorescence when all PS II reaction centers are dosed in dark- and light-acclimated cells, respectively - F_o fluorescence when all PS II reaction centers are open in dark acclimated cells - F_v variable fluorescence after dark acclimation (F_m–F_o)     

Plankton and hydrochemistry of Lake Futalaufquen (Patagonia,Argentina) during the growing season

Plankton communities and hydrochemistry of an oligotrophic lake occupying a glacial valley in Argentinian Patagonia (42 °49S; 71 °43W) were studied. Monthly samples at three stations integrated from 0 to 50 m and stratified samples at the site of maximum depth, were taken during the growing season. Transparency was always controlled by glacial silt, and not by phytoplankton. Lake water belongs to the calcium-bicarbonate type, with low conductivity (24 µS cm^–1), and poor buffering capacity. Forty-five phytoplankton taxa were found. Mean phytoplankton density was 49 cells ml^–1 and mean biomass 69 µg l^–1. N:P relationships, inorganic nitrogen exhaustion in the photic layer, and correlations between nutrients and phytoplankton density suggests nitrogen as the main limiting factor. Fifteen zooplankton species were found. Mean zooplankton density was 12.2 ind. l^–1 and mean biomass 22.9 µg l^–1. Diatoms and Boeckellidae were the dominant planktonic groups. Morphometry and hydrological factors were responsible for horizontal heterogeneity in phytoplankton and chemical variables.     

Partial purification and biochemical characterization of alkaline 5'-phosphodiesterase from barley malt sprouts

An alkaline 5-phosphodiesterase (5-PDE) from barley (Hordeum distichum) malt sprouts was partially purified by thermal treatment and acetone precipitation to diminish phosphomonoesterase (PME) activity. 5-PDE was purified 40-fold to a specific activity of 30 U mg^–1 protein with a final yield of about 32%. With synthetic substrate, the enzyme had an optimum pH of 8.9, maximum activity at 70 °C over 10 min, and a Km of 0.26 mM. The partially purified enzyme was activated by 10 mM Mg²⁺ up to 168% of the original activity, while Zn²⁺, Mn²⁺ and Cu²⁺ ions, chelating agent (EDTA) and NaN₃ (1–10 mM), and 5-ribonucleotides (1–5 mM) were inhibitory. Final enzyme preparation was stable over 8 d at 4 °C), at 70 °C for up to 120 min and without loss of activity over 90 d at –18 °C.