Assessment of Lemna gibba L. (duckweed) as a potential ecological indicator for contaminated aquatic ecosystem by boron mine effluent

Duckweeds, as a group, are important early warning indicators for the assessment of contaminated ecosystems due to their propensity to accumulate pollutants. In the present study, we investigated the potential use of Lemna gibba L. (Lemnaceae) as an ecological indicator for boron (B) mine effluent containing B concentration above 10 mg l⁻¹. For this purpose, L. gibba fronds were grown for 7 days in simulated water contaminated with B mine effluent. The important note is that this study was carried out in Kırka (Eskişehir, Turkey) B reserve area, which is the largest borax reserve in all over the world, under natural climatic conditions in the field. The results demonstrated that accumulations of B by L. gibba gradually increased based on the initial B concentrations (10, 25, 50, 100, and 150 mg l⁻¹) of the mine effluent. B concentration in the dry weight of the plant reached 639 mg kg⁻¹ when the minimum initial dosage (10 mg l⁻¹) was applied and 2711 mg kg⁻¹ when the maximum initial dosage (150 mg l⁻¹) was applied during the study. However, significant reductions in their relative growth rates occurred in 50, 100 and 150 mg l⁻¹ initial B concentrations. Results suggest that 25 mg l⁻¹ B concentration in water seemed to be a sensitive endpoint for L. gibba that could be used as a critical bioindicator level of B contaminated water. Following our data, we also constructed a simple growth model under the climatic conditions in this region of Turkey, but in instructive as a worldwide model. L. gibba is, therefore, suggested to be able to use as both an indicator and a phytoremediation tool because of its high accumulation capacity for B contaminated water.     

Protocorm culture of Dendrobium candidum in balloon type bubble bioreactors

Protocorm cultures of Dendrobium candidum were established in balloon type bubble bioreactors using Murashige and Skoog (MS) medium with 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5% (w/v) sucrose, 5:25 mM NH₄:NO₃ and 1% (v/v) banana homogenate for the production of biomass and bioactive compounds. In 3 l bioreactor containing 2 l medium, a maximum protocorm biomass (21.0 g l⁻¹ dry biomass) and also optimum quantities of total polysaccharides (389.3 mg g⁻¹ DW), coumarins (18.0 mg g⁻¹ DW), polyphenolics (11.9 mg g⁻¹ DW), and flavonoids (4.5 mg g⁻¹ DW) were achieved after 7 weeks of culture. Based on these studies, 5 and 10 l bioreactor cultures were established to harvest 80 g and 160 g dry biomass. In 10 l bioreactors, the protocorms grown were accumulated with optimal levels of polysaccharides (424.1 mg g⁻¹ DW), coumarins (15.8 mg g⁻¹ DW), polyphenols (9.03 mg g⁻¹ DW) and flavonoids (4.7 mg g⁻¹ DW). The bioreactor technology developed here will be useful for the production of important bioactive compounds from D. candidum.     

Mathematical analysis of trickling filter response to pentachlorophenol shock load

The response of a laboratory trickling filter to a step increase in pentachlorophenol (PCP) feed concentration was analyzed using continuous stirred tank (CSTR) and plug flow reactor (PFR) models. The CSTR model provided a slightly better fit to experimental data than the plug flow model when specific growth rate, μ, and PCP-degrading biomass concentration before the shock load, X₀, were variable parameters but was clearly superior when the mean residence time, τ, was added as a third parameter. The three-parameter CSTR model accurately represented six of seven concentration response curves corresponding to step increases in PCP feed concentration of 12–165 mg l⁻¹ and 20–150 mg l⁻¹. The continuing improvement in system response to repetitive 20–150 mg l⁻¹ shock loads was reflected by a monotonic increase in the optimal estimates of initial rate of biomass production.     

Treatment of dye-rich wastewater by an immobilized thermophilic cyanobacterial strain: Phormidium sp.

The removal of Remazol Blue and Reactive Black B by the immobilized thermophilic cyanobacterial strain Phormidium sp. was investigated under thermophilic conditions in a batch system, in order to determine the optimal conditions required for the highest dye removal. In the experiments, performed at pH 8.5, with different initial dye concentrations between 9.1 mg l⁻¹ and 82.1 mg l⁻¹ and at 45 °C, calcium alginate immobilized Phormidium sp. showed high dye decolorization, with maximum uptake yields ranging from 50% to 88% at all dye concentrations tested. When the effects of high dye concentrations on dye removal were investigated, the highest uptake yield in the beads was 50.3% for 82.1 mg l⁻¹ Remazol Blue and 60.0% for 79.5 mg l⁻¹ Reactive Black B. The highest color removal was detected at 45 °C and 50 °C incubation temperatures for all dye concentrations. As the temperature decreased, the removal yield of immobilized Phormidium sp. also decreased. At about 75 mg l⁻¹ initial dye concentrations, the highest specific dye uptake measured was 41.29–41.17 mg g⁻¹ for Remazol Blue and 47.69–43.82 mg g⁻¹ for Reactive Black B at 45 °C and 50 °C incubation temperatures, respectively, after 8 days incubation.     

Effects of Yucca schidigera extract on fermentation and degradation of steroidal saponins in the rumen simulation technique (RUSITEC)

A rumen simulation technique (RUSITEC) apparatus with eight 940 ml fermentation vessels was used to study the effects of the steroidal saponins in Yucca schidigera extract (YE) on ruminal microbial activity and saponin degradation. The YE contained approximately 4.4% (w/w) saponin, as smilagenin equivalents, and was included at 0 (control) or 0.5 mg ml⁻¹ (n=4) in the McDougall's buffer infused continuously into the vessels (dilution rate=0.75 day⁻¹). Each vessel received 5 g chopped alfalfa hay and 5 g concentrate (as-fed basis) daily for 22 days. Ammonia concentrations were lower (P<0.05) in effluent from vessels receiving YE than from controls for the first half of the study, but did not differ thereafter. Total amounts of VFA in effluent were not affected (P>0.05) by YE, but molar proportions of iso-butyric and iso-valeric acids were lower (P<0.05) in the YE vessels than in the controls in the first half of the experiment. Yucca extract at 0.5 mg ml⁻¹ did not affect (P>0.05) dry matter disappearance (DMD) from hay or from concentrate, nor did it affect total gas or methane production, or bacterial numbers (total or cellulolytic populations) in homogenates prepared from fermenter vessel liquid and feed particles. Protozoal numbers in the homogenates were substantially reduced (P<0.01) by YE (at 0.5 mg ml⁻¹), protease activity was increased (P<0.05), deaminase activity and activity against Ala₂ were unaffected (P>0.05) and activity against Ala₅ was reduced by 25% (P>0.05). When the homogenates from control and YE-supplemented (0.5 mg ml⁻¹) vessels were used to inoculate roll tubes containing 0 or 5 mg ml⁻¹ of YE, fewer colonies developed (P<0.01) in roll tubes containing YE than in those without YE, irrespective of the source of inoculum. Homogenates were also assayed for saponin degradation and for protease, peptidase and deaminase activities. Inoculum from the vessels receiving YE degraded saponin slightly during a 2 h incubation. Yucca extract at 0.5 mg ml⁻¹ altered proteolytic activity and reduced protozoal numbers, but did not affect DMD or bacterial activity, and did not induce resistance to YE at a concentration of 5 mg ml⁻¹.     

Bioavailability and extraction of heavy metals from contaminated soil by Atriplex halimus

Pot experiments were performed to evaluate the phytoremediation capacity of plants of Atriplex halimus grown in contaminated mine soils and to investigate the effects of organic amendments on the metal bioavailability and uptake of these metals by plants. Soil samples collected from abandoned mine sites north of Madrid (Spain) were mixed with 0, 30 and 60 Mg ha⁻¹ of two organic amendments, with different pH and nutrients content: pine-bark compost and horse- and sheep-manure compost. The increasing soil organic matter content and pH by the application of manure amendment reduced metal bioavailability in soil stabilising them. The proportion of Cu in the most bioavailable fractions (sum of the water-soluble, exchangeable, acid-soluble and Fe–Mn oxides fractions) decreased with the addition of 60 Mg ha⁻¹ of manure from 62% to 52% in one of the soils studied and from 50% to 30% in the other. This amendment also reduced Zn proportion in water-soluble and exchangeable fractions from 17% to 13% in one of the soils. Manure decreased metal concentrations in shoots of A. halimus, from 97 to 35 mg kg⁻¹ of Cu, from 211 to 98 mg kg⁻¹ of Zn and from 1.4 to 0.6 mg kg⁻¹ of Cd. In these treatments there was a higher plant growth due to the lower metal toxicity and the improvement of nutrients content in soil. This higher growth resulted in a higher total metal accumulation in plant biomass and therefore in a greater amount of metals removed from soil, so manure could be useful for phytoextraction purposes. This amendment increased metal accumulation in shoots from 37 to 138 mg pot⁻¹ of Cu, from 299 to 445 mg pot⁻¹ of Zn and from 1.8 to 3.7 mg pot⁻¹ of Cd. Pine bark amendment did not significantly alter metal availability and its uptake by plants. Plants of A. halimus managed to reduce total Zn concentration in one of the soils from 146 to 130 mg kg⁻¹, but its phytoextraction capacity was insufficient to remediate contaminated soils in the short-to-medium term. However, A. halimus could be, in combination with manure amendment, appropriate for the phytostabilization of metals in mine soils.     

Influence on dithiolopyrrolone antibiotic production by organic acids in Saccharothrix algeriensis NRRL B-24137

The influence of organic acids on growth and dithiolopyrrolone antibiotic production by Saccharothrix algeriensis NRRL B-24137 was studied. The production of dithiolopyrrolones depends upon the nature and concentration of the organic acids in the culture medium. Study of the nature of organic acids showed that the most effective organic acids for thiolutin specific production were maleic, 4-hydroxybenzoic, benzentetracarboxylic, pantothenic, pivalic and pyruvic acids (which yielded almost five-fold over the starting medium) and pimelic acid (more than three-fold). 4-Bromobenzoic acid showed the best production of senecioyl-pyrrothine (59 mg g⁻¹ DCW). Tiglic acid showed the best production of tigloyl-pyrrothine (22 mg g⁻¹ DCW). The highest yield of isobutyryl-pyrrothine (7.6 mg g⁻¹ DCW) was observed in the presence of crotonic acid. Sorbic acid yielded the best production of butanoyl-pyrrothine (26 mg g⁻¹ DCW). Methacrylic, butyric, pyruvic and 4-bromobenzoic acids also exhibited the best production of butanoyl-pyrrothine (27–11-fold).Study of organic acid concentration showed that among the selected organic acids, pimelic acid yielded the highest specific production of thiolutin (91 mg g⁻¹ DCW) at 7.5 mM; and senecioyl-pyrrothine (11 mg g⁻¹ DCW), tigloyl-pyrrothine (9 mg g⁻¹ DCW) and butanoyl-pyrrothine (3.5 mg g⁻¹ DCW) at 5 mM. Pyruvic acid at 1.25 mM enhanced the production of senecioyl-pyrrothine (4.3 mg g⁻¹ DCW). The maximum production of tigloyl-pyrrothine (18.6 mg g⁻¹ DCW) was observed in the presence of tiglic acid at 2.5 mM. Maximum production of isobutyryl-pyrrothine was observed in the presence of 7.5 mM tiglic acid. In addition, methacrylic acid (at 5 mM) and butyric acid (at 2.5 mM) enhanced the production of butanoyl-pyrrothine (26 and 20 times, respectively).The above results can be employed in the optimisation of the culture medium for the production of dithiolopyrrolone in higher quantities.     

Selenium uptake,speciation and stressed response of Nicotiana tabacum L.

The research on the function and mechanism of selenium (Se) is of great significance for the development of Se-enriched agricultural products. In this paper, uptake, speciation distribution, the effects on the flue-cured tobacco growth and antioxidant system of Se at different levels (0–22.2 mg Se kg⁻¹) were studied through a pot experiment, aiming to clarify flue-cured tobacco's response to Se stress and the relationship between Se speciation and antioxidant system. The results showed that the leaf area and number, the biomass and the chlorophyll content reached the maximum at 4.4 mg kg⁻¹ of Se treatment. Selenium at low levels (≤4.4 mg kg⁻¹) stimulated the growth of flue-cured tobacco by elevating the capability of antioxidant stress and reducing the malondialdehyde (MDA) content to 0.6–0.8 times of that of the control. However, high Se levels (≥11.1 mg kg⁻¹) depressed the capability of antioxidant stress and raised the MDA content to 1.5-fold of that of the control, and meanwhile the biomass of the aboveground parts and underground parts declined notably. The Se content in different parts of flue-cured tobacco significantly increased with the growth of Se levels. The range of Se content in roots, leaves and stems at 2.2–22.2 mg kg⁻¹ of Se treatment were 16.7–58.6 mg kg⁻¹, 2.6–37.3 mg kg⁻¹ and 2.2–10.3 mg kg⁻¹, respectively. According to the detection of different Se speciation, only selenocysteine (SeCys) was detectable in leaves at 2.2 mg kg⁻¹ Se treatment; SeCys, selenite [Se(IV)]and selenate [Se(VI)] were detected in flue-cured tobacco leaves at Se treatment (≥4.4 mg kg⁻¹), which accounted for 4.6–10%, 9–18.7% and 71–86% respectively; SeCys, selenomethionine (SeMet) and Se(IV) were detected in roots, and organic selenium(66–84%) was the main Se species at Se ≤ 11.1 mg kg⁻¹ treatment; four Se species [SeCys, SeMet, Se(IV) and Se(VI)] were detected in flue-cured tobacco roots, and the main Se species was inorganic Se (60%) at 22.2 mg kg⁻¹ Se treatment. That was to say, the percentage of organic Se species (SeCys and SeMet in flue-cured tobacco leaves and root) declined, whereas the ratio of inorganic Se species [Se(IV) and Se(VI)] increased with the growth of Se levels. The correlation analysis showed that the superoxide dismutase (SOD) activity as well as the glutathione (GSH) and MDA contents were positively correlated with the Se(IV) and Se(VI) contents at P < 0.01 and excessive inorganic Se might destruct the reactive oxygen species (ROS) balance and enhance the MDA content, thus causing damage to the plant growth. In a word, the present study suggested that the ratio of inorganic Se [Se(IV) and Se(VI)] was closely related with the growth and the antioxidant capacity of flue-cured tobacco and the excessive application of Se led to the higher proportion of inorganic Se and poorer antioxidant capacity, which ultimately inhibited the growth of flue-cured tobacco.     

Purification and characterization of an exo-polygalacturonase produced by Penicillium viridicatum RFC3 in solid-state fermentation

The pectinolytic enzyme obtained from Penicillium viridicatum RFC by solid-state fermentation was purified to homogeneity by pretreatment with kaolin (40 mg mL⁻¹) and ultrafiltration, followed by chromatography on a Sephadex G50 column. The apparent molecular weight of the enzyme was 24 kDa. Maximal activity occurred at pH 6.0 and at 60 °C. The enzyme proved to be an exo-polygalacturonase, releasing galacturonic acid by hydrolysis of highly esterified pectin. The presence of 10 mM Ba²⁺ increased the enzyme activity by 96% and its thermal stability by 30%, besides increasing its stability at acid pH. The apparent K_m with apple pectin as substrate was 1.82 mg mL⁻¹ and the V_max was 81 μmol min⁻¹ mg⁻¹.     

Multiple regeneration pathways in Spathoglottis plicata Blume — A study in vitro

Asymbiotic germination of immature seeds (embryos), and mature seeds and micropropagation of Spathoglottis plicata were described. Effects of three nutrition media namely, Murashige & Skoog (MS); Phytamax (PM); and Phyto-Technology orchid seed sowing medium (P₇₂₃), two carbon sources such as glucose and sucrose at 2–3% (w/v), two plant growth regulators such as 6-benzylaminopurine (BAP; 0.5–3.0 mg l^− 1) and α-naphthalene acetic acid (NAA; 0.5–2.0 mg l^− 1) and peptone (2.0 g l^− 1) were examined on seed germination, early protocorm development and micropropagation. The maximum germination of mature seeds (95%) was recorded in PM medium supplemented with 2% (w/v) sucrose + 2.0 g l^− 1 peptone. For germination of embryos P₇₂₃ medium supplemented with 1.0 mg l^− 1 BAP proved best. Multiple shoot buds or protocorm-like bodies (PLBs) were produced from stem segments of in vitro raised seedlings. Both direct organogenesis and embryogenesis were observed and the morphogenetic response was initiated by different concentrations and combinations of PGRs. The optimum PGR combination for maximal PLB regeneration was 1.0 mg l^− 1 NAA + 2.5 mg l^− 1 BAP, while 1.0 mg l^− 1 NAA + 1.0 mg l^− 1 BAP for shoot bud development. Strong and stout root system was induced in half strength PM medium supplemented with 0.5 mg l^− 1 IAA. The well-rooted plantlets were transferred to pots containing a potting mixture composed of saw dust, coconut coir, humus, and coal pieces at 1:1:1:2 (w/w) with 80% survival in outside environment and flowered after two years of transfer.     

Freeze tolerance and accumulation of cryoprotectants in the enchytraeid Enchytraeus albidus (Oligochaeta) from Greenland and Europe

The freeze tolerance and accumulation of cryoprotectants was investigated in three geographically different populations of the enchytraeid Enchytraeus albidus (Oligochaeta). E. albidus is widely distributed from the high Arctic to temperate Western Europe. Our results show that E. albidus is freeze tolerant, with freeze tolerance varying extensively between Greenlandic and European populations. Two populations from sub Arctic (Nuuk) and high Arctic Greenland (Zackenberg) survived freezing at −15 °C, whereas only 30% of a German population survived this temperature. When frozen, E. albidus responded by catabolising glycogen to glucose, which likely acted as a cryoprotectant. The average glucose concentrations were similar in the three populations when worms were frozen at −2 °C, approximately 50 μg glucose mg⁻¹ tissue dry weight (DW). At −14 °C the glucose concentrations increased to between 110 and 170 μg mg⁻¹ DW in worms from Greenland. The average glycogen content of worms from Zackenberg and Nuuk were about 300 μg mg⁻¹ DW, but only 230 μg mg⁻¹ DW in worms from Germany showing that not all glycogen was catabolised during the experiment. Nuclear magnetic resonance spectrometry (NMR) was used to screen for other putative cryoprotectants. Proline, glutamine and alanine were up regulated in frozen worms at −2 °C but only in relatively small concentrations suggesting that they were of little significance for freeze survival. The present study confirms earlier reports that freeze tolerant enchytraeids, like other freeze tolerant oligochaete earthworms, accumulate high concentrations of glucose as a primary cryoprotectant.     

A high pressure liquid chromatography-based assay for glutathione-S-transferase class distinction assay

In order to expedite the process of classification of the members of the family of glutathione-S-transferases (GSTs) high performance liquid chromatography with photodiode array detection (HPLC-PDA) was used as a means for measuring enzymatic activity. The GST chosen for the development of the HPLC-PDA technique was from equine liver (E-GST). The characterizing substrates, ethacrynic acid (EA) and bromosulfophthalein (BSP), along with previously gathered characterization data allowed for the distinction of α, μ or π-class enzymes. In an initial characterization of the previously unclassified E-GST it was determined that the enzyme was of the π-class with specific activities of 0.062, ± 0.0015 μmol min^− 1 mg^− 1 and 0.0019, ± 0.00064 μmol min^− 1 mg^− 1 for EA and BSP, respectively. Finally, the activity of the E-GST with the EA and BSP substrates, was measured by HPLC-PDA, and was found to be 0.027, ± 0.003 μmol min^− 1 mg^− 1 and 0.002, ± 0.0005 μmol min^− 1 mg^− 1, respectively. While the HPLC-PDA data do not mirror the spectrophotometric results quantitatively the overall response by the E-GST was the same. In general, the E-GSTs were shown to belong to the π-class when characterized by HPLC-PDA due to an EA specific activity greater than 0.01 μmol min^− 1 mg^− 1 and a negligible BSP activity (≤ 0.002 μmol min^− 1 mg^− 1).     

Chromate reduction by resting cells of Achromobacter sp. Ch-1 under aerobic conditions

Chromate reduction was carried out by resting cells of Achromobacter sp. Ch-1 with lactate as electron donor under aerobic conditions. The reduction activity of the samples supplemented with lactate was two times as those without lactate. The reduction rate was influenced by initial pH and lactate concentration. Under the optimal conditions, pH 9.0 and 4000 mg l⁻¹ lactate supplement, reduction rate was 5.45 mg l⁻¹ min⁻¹. The reduction rate decreased with increasing of Cr(VI) concentrations and increased with cell densities proportionally. The maximum reduction limit of Ch-1 cells was obtained at 2107 mg l⁻¹ of Cr(VI).     

Biosorption kinetics and isotherm studies of Acid Red 57 by dried Cephalosporium aphidicola cells from aqueous solutions

Equilibrium, kinetics and thermodynamic studies on the removal of Acid Red 57 (AR57) by biosorption onto dried Cephalosporium aphidicola (C. aphidicola) cells have been investigated in a batch system with respect to pH, contact time and temperature. The results showed that the equilibrium time was attained within 40 min and the maximum biosorption capacity of AR57 dye onto C. aphidicola cells was 2.08 × 10⁻⁴ mol g⁻¹ or 109.41 mg g⁻¹ obtained after contact with 0.4 g dm⁻³ biosorbent concentration, pH₀ of 1 and at a temperature of 20 °C. The pseudo-second-order kinetic model was observed to provide the best correlation of the experimental data among the kinetic models studied. Biosorption isotherm models were developed and the Langmuir, Freundlich and Dubinin–Radushkevich (D–R) isotherm models were conformed well to the experimental data. The changes of free energy, enthalpy and entropy of biosorption were also evaluated for the biosorption of AR57 dye onto C. aphidicola cells.     

Comparison of two matrices for selective recovery of C595 diabody fragment (dbFv) from Escherichia coli lysates

A comparative study of the performance of two types of adsorbent (Streamline Quartz Base and Upfront Matrices), derivatized with the same affinity ligand (RPAP) to recover C595 diabody fragment (dbFv) from Escherichia coli lysates has been undertaken. Both streamline and Upfront Matrices are characterized by a particle size range of 100–300 μm. Streamline has a density of 1.20 g cm⁻³ and ligand concentration of 0.85 μmol ml⁻¹. Upfront has a density of 1.35 g cm⁻³ and ligand concentration of 0.83 μmol ml⁻¹. The release of C595 dbFv from E. coli cells was achieved by a chemical lysis method. The recovery performance of both adsorbents was evaluated in terms of operational productivity and elution yield of C595 dbFv in packed bed (clarified feedstock) and expanded bed (unclarified and clarified feedstock) chromatography systems. Streamline and Upfront adsorbents exhibited diabody operational productivities of 131 and 202 mg l⁻¹, respectively, with an elution yield of 92 and 94%, respectively, in packed bed operation. Streamline and Upfront adsorbents exhibited diabody operational productivities of 54.5 and 123.7 mg l⁻¹, respectively, with an elution yield of 89 and 92%, respectively, in expanded bed operation.     

In vivo therapeutic potentiality of red seaweed,Asparagopsis (Bonnemaisoniales,Rhodophyta) in the treatment of Vibriosis in Penaeus monodon Fabricius

The crude extract of the red seaweed, Asparagopsis sp. was evaluated for in vivo antibacterial activity against the shrimp vibrio pathogens. The algal extract was rationalized with commercial shrimp feed and orally administered for different duration of time followed by the artificial bacterial challenge experiment. In dose titration experiments, the oral administration of Asparagopsis sp. at a dosage of 850 mg kg^–1 of biomass was highly efficacious in the treatment of natural infestations of Vibriosis in Penaeus monodon. The results of the confirmatory dose experiment revealed that the prophylactic treatment with moderate dose of 850 mg kg^–1 of biomass day^–1 for four weeks followed by 14 days of post infection therapy was highly effective in controlling Vibrio infection in shrimps. Moreover, results of the percent survival index and microbiological analysis clearly show that Asparagopsis extract incorporated medicated feed had broad therapeutic potential for managing shrimp Vibriosis. In addition, in vivo trials and results obtained in this work are based on the crude organic extract sourced from an unidentified Asparagopsis cryptic lineage, therefore further molecular analysis to identify the species will be required.     

Growth of the fungus Paecilomyces lilacinus with n-hexadecane in submerged and solid-state cultures and recovery of hydrophobin proteins

The filamentous fungus Paecilomyces lilacinus was grown on n-hexadecane in submerged (SmC) and solid-state (SSC) cultures. The maximum CO₂ production rate in SmC (V_max = 11.7 mg CO₂ L_g⁻¹ day⁻¹) was three times lower than in SSC (V_max = 40.4 mg CO₂ L_g⁻¹ day⁻¹). The P. lilacinus hydrophobin (PLHYD) yield from the SSC was 1.3 mg PLHYD g protein⁻¹, but in SmC, this protein was not detected. The PLHYD showed a critical micelle concentration of 0.45 mg mL⁻¹. In addition, the PLHYD modified the hydrophobicity of Teflon from 130.1 ± 2° to 47 ± 2°, forming porous structures with some filaments <1 μm and globular aggregates <0.25 μm diameter. The interfacial studies of this PLHYD could be the basis for the use of the protein to modify surfaces and to stabilize compounds in emulsions.     

Improvement of succinate production by overexpression of a cyanobacterial carbonic anhydrase in Escherichia coli

Succinate fermentation was investigated in Escherichia coli strains overexpressing cyanobacterium Anabaena sp. 7120 ecaA gene encoding carbonic anhydrase (CA). In strain BL21 (DE3) bearing ecaA, the activity of CA was 21.8 U mg⁻¹ protein, whereas non-detectable CA activity was observed in the control strain. Meanwhile, the activity of phosphoenolpyruvate carboxylase (PEPC) increased from 0.2 U mg⁻¹ protein to 1.13 U mg⁻¹ protein. The recombinant bearing ecaA reached a succinate yield of 0.39 mol mol⁻¹ glucose at the end of the fermentation. It was 2.1-fold higher than that of control strain which was just 0.19 mol mol⁻¹ glucose. EcaA gene was also introduced into E. coli DC1515, which was deficient in glucose phosphotransferase, lactate dehydrogenase and pyruvate:formate lyase. Succinate yield can be further increased to 1.26 mol mol⁻¹ glucose. It could be concluded that the enhancement of the supply of HCO₃⁻ in vivo by ecaA overexpression is an effective strategy for the improvement of succinate production in E. coli.     

Nitrite inhibition and intermediates effects on Anammox bacteria: A batch-scale experimental study

A batch test procedure, based on manometric measurements, was used to study the Anammox process, in particular the inhibition due to nitrite and the effects of hydroxylamine and hydrazine, indicated as possible intermediates of the process. The maximum nitrite removal rate (MNRR) was measured. The method showed good reliability with a standard error of 4.5 ± 3.3% (n: 41). All the tests were carried out on samples taken from a pilot plant with Anammox suspended biomass. The tests were used also to monitor the reactor activity. By testing different spiked additions of nitrite (10–75 mg NO₂⁻-N L⁻¹), a short-term inhibition, with more than 25% MNRR decrease, was found at concentrations higher than 60 mg NO₂⁻-N L⁻¹. Repeated additions of nitrite higher than 30 mg NO₂⁻-N L⁻¹ caused losses of activity. After a complete loss of activity, spiked additions of hydroxylamine (30 mg N L⁻¹ in total) determined a 20% permanent recovery. Low amounts of the intermediates (1–3 mg N L⁻¹) applied on partially inhibited samples and uninhibited samples produced temporary increases in activity up to 50% and 30%, respectively.     

Immobilized glucose oxidase on different supports for biotransformation removal of glucose from oligosaccharide mixtures

Four immobilized forms of glucose oxidase (GOD) were used for biotransformation removal of glucose from its mixture with dextran oligosaccharides. GOD was biospecifically bound to Concanavalin A-bead cellulose (GOD-ConA-TBC) and covalently to triazine-bead cellulose (GOD-TBC). Eupergit C and Eupergit CM were used for preparation of other two forms of immobilized GOD: GOD-EupC and GOD-EupCM. GOD-ConA-TBC and GOD-EupC exhibited the best operational and storage stabilities. pH and temperature optima of these two immobilized enzyme forms were broadened and shifted to higher values (pH 7 and 35 °C) in comparison with those of free GOD. The decrease of V_max values after immobilization was observed, from 256.8 ± 7.0 μmol min⁻¹ mg_GOD⁻¹ for free enzyme to 63.8 ± 4.2 μmol min⁻¹ mg_GOD⁻¹ for GOD-ConA-TBC and 45 ± 2.7 μmol min⁻¹ mg_GOD⁻¹ for GOD-EupC, respectively. Depending on the immobilization mode, the immobilized GODs were able to decrease the glucose content in solution to 3.8–15.6% of its initial amount The best glucose conversion, was achieved by an action of GOD-EupCM on a mixture of 100 g dextran with 9 g of glucose (i.e. 98.7% removal of glucose).